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As a robust antagonist of myostatin (MSTN), follistatin (FST) is an important regulator of skeletal muscle development, and the delivery of FST to muscle tissue represents a potential therapeutic strategy for muscular dystrophies. The N terminus and FSI domain of FST are the functional domains for MSTN binding. Here, we aimed to achieve site-specific integration of FSI-I-I, including the signal peptide, N terminus, and three FSI domains, into the last codon of the porcine MSTN gene using a homology-mediated end joining (HMEJ)-based strategy mediated by CRISPR-Cas9. Based on somatic cell nuclear transfer (SCNT) technology, we successfully obtained FSI-I-I knockin pigs. H&E staining of longissimus dorsi and gastrocnemius cross-sections showed larger myofiber sizes in FSI-I-I knockin pigs than in controls. Moreover, the Smad and Erk pathways were inhibited, whereas the PI3k/Akt pathway was activated in FSI-I-I knockin pigs. In addition, the levels of MyoD, Myf5, and MyoG transcription were upregulated while that of MRF4 was downregulated in FSI-I-I knockin pigs. These results indicate that the FSI-I-I gene mediates skeletal muscle hypertrophy through an MSTN-related signaling pathway and the expression of myogenic regulatory factors. Overall, FSI-I-I knockin pigs with hypertrophic muscle tissue hold great promise as a therapeutic model for human muscular dystrophies., Graphical abstract, The FSI-I-I, including the signal peptide, N terminus, and three FSI domains of FST, were site-specifically integrated into the last codon of the MSTN gene in porcine using a HMEJ-based strategy mediated by CRISPR-Cas9. The FSI-I-I knockin pigs obtained based on somatic cell nuclear transfer exhibited increased skeletal muscle mass.

Using the combined application of L-arginine with HMG-Co-A reductase inhibitors simvastatin, atorvastatin, rosuvastatin and nanoparticulated rosuvastatin on the background of modeling of sepsis-induced disease through the introduction of strain 603 Staphylococcus aureus shows endotelio- and cardioprotective effects, manifesting itself in preventing the proliferation of endothelial dysfunction coefficient (CED), adrenoreactivity, maintenance of myocardial reserve and the normalization of biochemical markers values (Total NO, eNOS expression, C-reactive protein, IL-6, TNF). In this case, the combined therapy was so effective that the values obtained thereunder did not differ from those obtained from control animals.

Experts of the Union of Pediatricians of Russia have developed modern guidelines on the management of children with acute obstructive laryngitis and epiglottitis. Croup is the most common cause of acute upper respiratory obstruction in children aged from 6 months to 6 years. Usually respiratory viruses are the pathogenic agents of the disease. The etiological factor of epiglottitis is Haemophilus influenzae type b in the vast majority of cases (> 90%). However, nowadays, epiglottitis caused by this pathogen agent is more common in adults in countries with mass immunization against hemophilic infection. This article considers in detail etiology, pathogenesis, and classification issues. Special attention is given to diagnosis, differential diagnosis, and therapy (via evidence-based approach) of acute obstructive laryngitis in children.

Urocortin-2 (Ucn-2) has demonstrated cardioprotective actions against myocardial ischemia-reperfusion (I/R) injuries. Herein, we explored the protective role of Ucn-2 through microRNAs (miRNAs) post-transcriptional regulation of apoptotic and pro-fibrotic genes. We determined that the intravenous administration of Ucn-2 before heart reperfusion in a Wistar rat model of I/R recovered cardiac contractility and decreased fibrosis, lactate dehydrogenase release, and apoptosis. The infusion of Ucn-2 also inhibited the upregulation of 6 miRNAs in revascularized heart. The in silico analysis indicated that miR-29a and miR-451_1∗ are predicted to target many apoptotic and fibrotic genes. Accordingly, the transfection of neonatal rat ventricular myocytes with mimics overexpressing miR-29a, but not miR-451_1∗, prevented I/R-induced expression of pro- and anti-apoptotic genes such as Apaf-1, Hmox-1, and Cycs, as well as pro-fibrotic genes Col-I and Col-III. We also confirmed that Hmox-1, target of miR-29a, is highly expressed at the mRNA and protein levels in adult rat heart under I/R, whereas, Ucn-2 abolished I/R-induced mRNA and protein upregulation of HMOX-1. Interestingly, a significant upregulation of Hmox-1 was observed in the ventricle of ischemic patients with heart failure, correlating negatively with the left ventricle ejection fraction. Altogether, these data indicate that Ucn-2, through miR-29a regulation, provides long-lasting cardioprotection, involving the post-transcriptional regulation of apoptotic and fibrotic genes., This study was co-financed by FEDER Funds [US-1381135], Agencia Estatal de Investigación [PID2019-104084GB-C22/AEI/10.13039/501100011033]; the Institute of Carlos III [PI18/01197; Red TerCel-Grant RD16/0011/0034]; the Andalusia Government [grant number: PI-0193-2018]; and by Agence National de la Recherche [ANR-19-14-0031-01].

Circular RNAs (circRNAs) have been proven to play key roles in the development and progression of various types of cancers. However, there were no reported studies on the roles of circRNA mediator complex subunit 27 (circMED27) in tumors including hepatocellular carcinoma (HCC). In this study, we found that circMED27 was significantly increased in HCC serum and that higher levels of circMED27 were correlated with bad clinical characteristics and poor prognoses of patients with HCC. Furthermore, upregulated circMED27 promoted HCC resistance to lenvatinib. Our mechanistic investigations revealed that circMED27 functions as a competing endogenous RNA (ceRNA) for miR-655-3p to upregulate ubiquitin-specific peptidase 28 (USP28) expression. Thus, we are led to conclude that circMED27 acts as a potential therapeutic target for HCC patients receiving lenvatinib therapy and may represent a promising molecular biomarker for forecasting lenvatinib-resistant HCC., Graphical abstract, Zhang et al. provide a promising biomarker and therapeutic target for the recovery sensitivity of HCC to lenvatinib. circMED27 was upregulated in both tumor tissues and serum of HCC patients. In addition, circMED27 could bind to miR-655-3p and positively regulate USP28 expression, further inducing lenvatinib resistance in HCC.

Clear cell renal cell carcinoma (ccRCC) is the most lethal urological cancer and is characterized by a high rate of metastasis and relapse. N6-Methyladenosine (m6A) is implicated in various stages of cancer development. However, a thorough understanding of m6A-modified lncRNAs in ccRCC is lacking. The results showed that METTL14 had decreased expression in ccRCC tissues. In addition, the expression of METTL14 was negatively correlated to the prognosis, stage, and ccRCC tumor grade. The silencing of METTL14 was shown to significantly increase metastasis in vitro and in vivo. High-throughput methylated RNA immunoprecipitation sequencing (MeRIP-seq) showed that the m6A levels of Lnc-LSG1 could be regulated by METTL14. Lnc-LSG1 can directly bind to ESRP2 protein and promote ESRP2 degradation via facilitating ESRP2 ubiquitination. However, m6A modification on Lnc-LSG1 can block the interaction between Lnc-LSG1 and ESRP2 via the m6A reader, YTHDC1. Taken together, our findings unraveled the novel mechanism of METTL14 inhibiting ccRCC progression, and explored the correlation between m6A and lncRNA in ccRCC for the first time., Graphical abstract, N6-Methyladenosine (m6A) is the most common RNA modification and implicated in various stages of cancer development. METTL14 is an important methyltransferase of m6A. For the first time, we provide insights into the function and mechanism of m6A-modified lncRNA in ccRCC and identify a “METTL14-YTHDC1-Lnc-LSG1” regulation axis in ccRCC progression.

Mesenchymal stromal cell (MSC) transplantation has been a promising therapeutic strategy for repairing heart tissues post-myocardial infarction (MI). Nevertheless, its therapeutic efficacy remains low, which is mainly ascribed to the low viability of transplanted MSCs. Recently, long noncoding RNAs (lncRNAs) have been reported to participate in diverse physiological and pathological processes, but little is known about their role in MSC survival. Using unbiased transcriptome profiling of hypoxia-preconditioned MSCs (HP-MSCs) and normoxic MSCs (N-MSCs), we identified a lncRNA named lung cancer-associated transcript 1 (LUCAT1) under hypoxia. LUCAT1 knockdown reduced the survival of engrafted MSCs and decreased the MSC-based therapeutic potency, as shown by impaired cardiac function, reduced cardiomyocyte survival, and increased fibrosis post-MI. Conversely, LUCAT1 overexpression had the opposite results. Mechanistically, LUCAT1 bound with and recruited jumonji domain-containing 6 (JMJD6) to the promoter of forkhead box Q1 (FOXQ1), which demethylated FOXQ1 at H4R3me2(s) and H3R2me2(a), thus downregulating Bax expression and upregulating Bcl-2 expression to attenuate MSC apoptosis. Therefore, our findings revealed the protective effects of LUCAT1 on MSC apoptosis and demonstrated that the LUCAT1-mediated JMJD6-FOXQ1 pathway might represent a novel target to potentiate the therapeutic effect of MSC-based therapy for ischemic cardiovascular diseases., Graphical Abstract, A novel lncRNA, LUCAT1, bound with and recruited JMJD6 to the promoter of FOXQ1, which demethylated FOXQ1 at H4R3me2(s) and H3R2me2(a), thus decreasing pro-apoptotic protein (Bax and Cleaved caspase-3) and increasing anti-apoptotic protein (Bcl-2) expression to attenuate MSC apoptosis. LUCAT1-mediated JMJD6-FOXQ1 pathway might be a promising pathway for improving MSCs’ therapeutic efficacy in MI.

Context: The validation of a method is synonymous with the quality of the obtained results. The dissolution test is an analytical technique to evaluate the quality and stability of drugs during their development. Aims: To evaluate whether the UV spectrophotometric method for the quantification of losartan potassium in tablets from the dissolution test at pH 1.2, 4.5 and 6.8 meet with the validation parameters. Methods: The determination and evaluation of validation parameters were carried out under the guidelines of the regulatory entities. Linearity and range, accuracy, precision, specificity, limits of detection and quantification, robustness, stability of the sample solution were evaluated, and the filter test was added. All data obtained were subject to an analysis of variance and t-student analysis with a confidence level of 95% (α = 0.05). Results: The UV spectrophotometric method meets the acceptance criteria for each validation parameter. Likewise, it was identified that the prepared solutions were stable at pH 6.8 for 24 hours; however, they were not stable at pH 1.2 and 4.5. Conclusions: The method meets with the validation criteria and is suitable to be used for quantifying samples obtained from the losartan potassium tablet dissolution test.

Diabetic retinopathy is a heterogeneous retinal degenerative disease with the microvascular dysfunction being recognized as a hallmark of the advanced stage. In this study, we demonstrated that exosomes collected from the vitreous humor of proliferative diabetic retinopathy patients promoted proliferation, migration and tube formation ability of primary human retinal endothelial cells via its elevated miR-9-3p expression level. Müller glia cells were further recognized as the sole source of the aberrantly expressed miR-9-3p, and both in vitro and in vivo experiments validated that Müller glia-derived exosomes aggravate vascular dysfunction under high glucose. Mechanistically, exosomal miRNA-9-3p was transferred to retinal endothelial cells and bound to the sphingosine-1-phosphate receptor S1P1 coding sequence, which subsequently activated VEGFR2 phosphorylation and internalization in the presence or absence of exogenous VEGF-A. We successfully orchestrated the dynamic crosstalk between retinal Müller glia cells and endothelial cells in pathological condition, which may provide a novel biomarker or promising therapeutic agents for the treatment of diabetic retinopathy., Graphical abstract, Liu et al. show that, in the vitreous humor of diabetic retinopathy patients, exosomal miRNA-9-3p secreted by Müller cells promote retinal angiogenesis by restricting sphingosine-1-phosphate receptor S1P1. This provides a better understanding on molecular mechanisms underlying diabetic retinopathy and sheds light on new potential therapeutic targets in future.

Evasion of immune destruction is a major hallmark of cancer. Recent US Food and Drug Administration (FDA) approvals of various immunomodulating therapies underline the important role that reprogramming the immune system can play in combating this disease. However, a wide range of side effects still limit the therapeutic potential of immunomodulators, suggesting a need for more precise reagents with negligible off-target and on-target/off-tumor effects. Aptamers are single-chained oligonucleotides that bind their targets with high specificity and affinity owing to their three-dimensional (3D) structures, and they are one potential way to address this need. In particular, bispecific aptamers (bsApts) have been shown to induce artificial immune synapses that promote T cell activation and subsequent tumor cell lysis in various in vitro and in vivo pre-clinical models. We discuss these advances here, along with gaps in bsApt biology at both the cellular and resident tissue levels that should be addressed to accelerate their translation into the clinic. The broad application, minimal production cost, and relative lack of immunogenicity of bsApts give them some ideal qualities for manipulating the immune system. Building upon lessons from other novel therapies, bsApts could soon provide clinicians with an immunomodulating toolbox that is not only potent and efficacious but exercises a wide therapeutic index.

Context: Several experimental results and clinical reports using Mangifera indica L extract (MSBE) suggest its potential utility in osteoarthritis (OA) mixed pain. Aims: To examine the possible therapeutic effects and safety of supplementation on osteoarthritis (OA) pain. Methods: Fifty patients with painful knee OA who had undergone a year of conventional treatment that included paracetamol and non-pharmacological therapies were randomly allocated to the experimental group (n = 21), which received a daily dose of 900 mg of extract supplementation or preceding usual treatment and placebo in the same form (n = 17) for a period of 120 days. The primary measure outcome was the change in the average daily pain diary score (ADPS) using the Likert scale. Also, a multidimensional measure of pain, stiffness and functional disability on The Western Ontario and Mc Master Universities (WOMAC) index for knee OA and ultrasonographic chronic signs of synovitis such as effusion and synovial thickness were evaluated. Results: Change from baseline in ADPS of the MSBE supplemented group showed a significant reduction after two weeks that lasted for 120 days with respect to the placebo group. Significant improvements in pain and functional disability WOMAC sub-scores, number of joints with synovial thickness and effusion after MSBE supplementation vs. placebo were observed. Non-adverse effects were reported in the experimental group. Conclusions: These results suggest that MSBE supplementation has a beneficial effect on OA pain and disability.

It is well established that natural killer (NK) cells can be used as an alternative candidate of T cells for adoptive cell therapy (ACT) due to its high killing capacity, off-the-shelf utility, and low toxicity. Though NK cells provide rapid and potent immune effects, they still suffer from insufficient infiltration and tumor immunosuppression environment, which result in unsatisfactory therapeutic efficiency. Herein, a highly stable CD16/PD-L1 bi-specific aptamer (defined as CP-bi-apt) with high affinity and selectivity was introduced to overcome these obstacles. This CP-bi-apt can mediate a significant antitumor immunity by recruiting CD16-positive NK cells to directly contact with PD-L1 high-expressed tumor cells. In addition, the induced up-regulation of PD-L1 on tumor cells can inevitably occur as an adaptive response to most of the immunotherapeutic strategies. The prepared CP-bi-apt can be further used as an immune checkpoint inhibitor to specifically bind to PD-L1, thus reducing the negative impact of PD-L1 over-expression on the therapeutic efficacy. Furthermore, this CP-bi-apt-based immunotherapy is simple, highly efficient, and has low side effects, showing a promising potential for clinical translation.

Context: Compounding sterile preparations in hospitals requires special attention because the process is more complex. Errors that occur when compounding sterile preparations can cause contamination of the prepared preparations. Aims: To evaluate the compounding of sterile preparations in a referral hospital. Methods: This research was conducted by observing nine critical aspects in the compounding of sterile preparations. The research instrument used was a checklist, as regulated in USP Guidelines and Basic Guidelines for Dispensing Sterile Preparations. After that, the most mixed preparations were tested for sterility using a fluid thioglycollate medium and soybean casein digest medium. Results: Of the 36 compounding sterile preparations, discrepancies were found in the aspects of compounding personnel (100%), building (100%), equipment (100%), aseptic procedures (100%), packaging (0%), labels (100%), storage (2.78%), distribution (2.78%) and quality assurance (100%). Furthermore, sterile preparations prepared in the cleanroom in this referral hospital indicate the occurrence of microbial contamination. Conclusions: Most of the critical aspects in compounding sterile preparations at this referral hospital have not met the recommendations of the USP Guidelines and the Basic Guidelines for the Dispensing of Sterile Preparations, except for the packaging aspect. Improvements in several critical aspects need to be made to ensure the quality of the prepared preparations from microbial contamination.

Genetic lineage tracing is indispensable to unraveling the origin, fate, and plasticity of cells. However, the intrinsic leakiness in the CreER-loxP system raises concerns on data interpretation. Here, we reported the generation of a novel dual inducible CreER-loxP system with superior labeling characteristics. This two-component system consists of membrane localized CreER (mCreER: CD8α-FRB-CS-CreER) and TEV protease (mTEVp: CD8α-FKBP-TEVp), which are fusion proteins incorporated with the chemically induced dimerization machinery. Rapamycin and tamoxifen induce sequential dimerization of FKBP and FRB, cleavage of CreER from the membrane, and translocation into the nucleus. The labeling leakiness in Ad293 cells reduced dramatically from more than 70% to less than 5%. This tight labeling feature depends largely on the association of mCreER with HSP90, which conceals the TEV protease cutting site between FRB and CreER and thus preventing uninduced cleavage of the membrane-tethering CreER. Membrane-bound CreER also diminished significantly cytotoxicity. Our studies showed mCreER under the control of the rat insulin promoter increased labeling specificity in MIN6 islet beta-cells. Viability and insulin secretion of MIN6 cells remained intact. Our results demonstrate that this novel system can provide more stringent temporal and spatial control of gene expression and will be useful in cell fate probing.

This study aimed to elucidate the role of nitric oxide (NO) in intestinal stem cells in methotrexate-induced ileal mucositis in rats. Methotrexate induced the mRNA expressions of the Wnt/β-catenin target genes Wnt3a, Sox9, and Lgr5 and the Wnt-antagonist gene sFRP-1 and the protein expressions of Lgr5 and sFRP-1. Methotrexate also induced Lgr5+ cells and lysozyme+ cells. A non-selective NO inhibitor inhibited the methotrexate induction of Wnt/β-catenin target genes and Lgr5+ cells but enhanced that of sFRP-1 expression. Thus, methotrexate mediates the integrity of intestinal stem cells partly through NO-dependent Wnt/β-catenin signaling and may enhance tolerability to methotrexate-induced injury.

Context: Alzheimer’s disease (AD) is one of the biggest causes of public health problems today, especially in older, where women have a greater risk of developing AD than men. However, the current AD therapy has not been satisfactory. Nigella sativa(NS) has bioactive compounds of flavonoid compounds and estrogenic effects that could be alternative medicine. Aims: To evaluate an extract of N. sativa on improving memory function through reducing GSK3β activation and Aβ42/40 ratio in a model of AD in rats. Methods: This research was an experimental study with the post-test-only group design using 24 Wistar rats divided into eight groups. Conjugated equine estrogen (CEE) 0.018-0.0036 mg/day and NS 100, 200, and 400 mg/kg BW were administered orally two weeks before intraperitoneal AlCl3 induction, then continued for up to 8 weeks. In the last five days of the study, the Morris water maze memory test was performed to analyze the expression of amyloid β (Ab), neurofibrillary tangles (NFT), glycogen synthase kinase 3β (GSK3β), and Aβ42/40 ratio. Results: There were significant differences in the memory test (p = 0.000), the expression Aβ40 (p = 0.000), Aβ42 (p = 0.000), NFT (p = 0.000) and GSK3β levels (p = 0.000). The lowest expression of Aβ40 in the NS100 group, Aβ42 in the NS400, NFT in the CEE group, and GSK3β levels in the NS100 group. Administration of NS increased the plasma ratio of Ab42/40 with the highest mean in the CEE group (p = 0.001). Conclusions: The administration of NS extract affected improving memory function by decreasing the expression of Aβ, NFT, GSK3β in ovariectomized Wistar rats supplemented with AlCl3.

Gene therapy offers great promises for a cure of hemophilia A resulting from factor VIII (FVIII) gene deficiency. We have developed and optimized a non-viral ultrasound-mediated gene delivery (UMGD) strategy. UMGD of reporter plasmids targeting mice livers achieved high levels of transgene expression predominantly in hepatocytes. Following UMGD of a plasmid encoding human FVIII driven by a hepatocyte-specific promoter/enhancer (pHP-hF8/N6) into the livers of hemophilia A mice, a partial phenotypic correction was achieved in treated mice. In order to achieve persistent and therapeutic FVIII gene expression, we adopted a plasmid (pHP-hF8-X10) encoding an FVIII variant with significantly increased FVIII secretion. By employing an optimized pulse-train ultrasound condition and immunomodulation, the treated hemophilia A mice achieved 25%-150% of FVIII gene expression on days 1-7 with very mild transient liver damage, as indicated by a small increase of transaminase levels that returned to normal within 3 days. Therapeutic levels of FVIII can be maintained persistently without the generation of inhibitors in mice. These results indicate that UMGD can significantly enhance the efficiency of plasmid DNA transfer into the liver. They also demonstrate the potential of this novel technology to safely and effectively treat hemophilia A.

Vascular calcification (VC), or calcium deposition inside the blood vessels, is common in patients with atherosclerosis, cardiovascular disease, and chronic kidney disease. Although several treatments are available to reduce calcification, the incidence of VC continues to rise. Recently, there have been several reports describing the regulation of circular RNAs (circRNAs) in various diseases. However, the role of circRNAs in VC has not yet been fully explored. Here, we investigated the function of circSmoc1-2, one of the circRNAs generated from the Smoc1 gene, which is downregulated in response to VC. CircSmoc1-2 is localized primarily to the cytoplasm and is resistant to exonuclease digestion. Inhibition of circSmoc1-2 worsens VC, while overexpression of circSmoc1-2 reduces VC, suggesting that circSmoc1-2 can prevent calcification. We went on to investigate the mechanism of circSmoc1-2 as a microRNA sponge and noted that miR-874-3p, the predicted target of circSmoc1-2, promotes VC, while overexpression of circSmoc1-2 reduces VC by suppressing miR-874-3p. Additionally, we identified the potential mRNA target of miR-874-3p as Adam19. In conclusion, we revealed that the circSmoc1-2/miR-874-3p/Adam19 axis regulates VC, suggesting that circSmoc1-2 may be a novel therapeutic target in the treatment of VC., Graphical abstract, Vascular calcification, or calcium deposition inside the blood vessels, is common in patients with atherosclerosis, cardiovascular disease, and chronic kidney disease. In this study, the authors demonstrate that the circSmoc1-2/miR-874-3p/Adam19 axis regulates vascular calcification, suggesting the circSmoc1-2 may be a novel therapeutic target in the treatment of vascular calcification.

Colorectal cancer (CRC) is the fourth leading cause of cancer-related mortality globally. Therefore, a better understanding of the early molecular events of this disease is needed. Long noncoding RNAs (lncRNAs) play a critical role in the regulation of tumorigenesis and cancer progression. In this study, we investigated the characteristics of ZFAS1 in CRC. We analyzed three independent microarray datasets of CRC tissues from GEO and found that ZFAS1 expression was remarkably upregulated in all three datasets. Moreover, we validated the overexpression of ZFAS1 in CRC tissues compared with normal tissues and found that ZFAS1 was positively correlated with tumor size and metastasis in CRC. Knockdown of ZFAS1 significantly suppressed the malignant phenotype and lipogenesis of CRC cells. Mechanistically, ZFAS1 binds polyadenylate-binding protein 2 (PABP2) to stabilize SREBP1 mRNA, thereby increasing the expression of SREBP1 and its target genes stearoyl-CoA desaturase (SCD1) and fatty acid synthase (FASN), thus promoting CRC lipid accumulation. These data demonstrated that ZFAS1 could act as an oncogene for CRC and that ZFAS1 reprograms lipid metabolism by binding with PABP2 to stabilize SREBP1 mRNA accumulation, implicating it as a novel and potent target for the treatment of CRC., Graphical abstract, Colorectal cancer (CRC) is a leading cause of cancer-related death worldwide. Liu et al. report that long noncoding RNA ZFAS1 regulates fatty acid synthesis, thereby providing survival advantages for malignant phenotype transformation of CRC, and may be a potential therapeutic target for CRC.

FilaminC (Flnc) is a member of the actin binding protein family, which is preferentially expressed in the cardiac and skeletal muscle tissues. Although it is known to interact with proteins associated with myofibrillar myopathy, its unique role in skeletal muscle remains largely unknown. In this study, we identify the biological functions of Flnc in vitro and in vivo using chicken primary myoblast cells and animal models, respectively. From the results, we observe that the growth rate and mass of the skeletal muscle of fast-growing chickens (broilers) were significantly higher than those in slow-growing chickens (layers). Furthermore, we find that the expression of Flnc in the skeletal muscle of broilers was higher than that in the layers. Our results indicated that Flnc was highly expressed in the skeletal muscle, especially in the skeletal muscle of broilers than in layers. This suggests that Flnc plays a positive regulatory role in myoblast development. Flnc knockdown resulted in muscle atrophy, whereas the overexpression of Flnc promotes muscle hypertrophy in vivo in an animal model. We also found that Flnc interacted with dishevelled-2 (Dvl2), activated the wnt/β-catenin signaling pathway, and controlled skeletal muscle development. Flnc also antagonized the LC3-mediated autophagy system by decreasing Dvl2 ubiquitination. Moreover, Flnc knockdown activated and significantly increased mitophagy. In summary, these results indicate that the absence of Flnc induces autophagy or mitophagy and regulates muscle atrophy., Graphical Abstract, Filamin C (Flnc) interacts with dishevelled-2 (Dvl2) to activate wnt/β-catenin signaling, consequently regulating skeletal muscle development. Furthermore, Flnc stabilizes Dvl2 by reducing the level of LC3-labeled Dvl2 and preventing cells from undergoing autophagy. In addition, Flnc silence-induced autophagy mediates muscle atrophy by mitochondrial clearance.

Lung cancer (LC) is the leading cause of cancer-related deaths worldwide, with high morbidity and mortality. Non-small cell lung cancer (NSCLC) is a major pathological type of LC and accounts for more than 80% of all cases. Circular RNAs (circRNAs) are a large class of non-coding RNAs (ncRNAs) with covalently closed-loop structures, a high abundance, and tissue-specific expression patterns. They participate in various pathophysiological processes by regulating complex gene networks involved in proliferation, apoptosis, migration, and epithelial-to-mesenchymal transition (EMT), as well as metastasis. A growing number of studies have revealed that the dysregulation of circRNAs contributes to many aspects of cancer progression, such as its occurrence, metastasis, and recurrence, suggesting their great potential as efficient and specific biomarkers in the diagnosis, prognosis, and therapeutic targeting of NSCLC. In this review, we systematically elucidate the characteristics, biogenesis, and functions of circRNAs and focus on their molecular mechanisms in NSCLC progression. Moreover, we highlight their clinical implications in NSCLC treatment., Graphical Abstract, In this review, Liu et al. systematically summarize the biological characteristics, biogenesis, and functions of circRNAs and focus on their mechanisms in non-small cell lung cancer (NSCLC) progression. Moreover, the authors also highlight the potential of circRNAs as diagnostic and prognostic biomarkers and therapeutic targets for NSCLC.

We identified a novel interactome, circ_0001018/miR-338-3p/SOX4, in papillary thyroid cancer (PTC), and we intended to confirm the regulatory relationship between the three and to study the effects of the three in PTC. The bioinformatics method was used to screen out the circular RNA and mRNA of interest. A cellular fractionation assay and fluorescence in situ hybridization (FISH) assay were conducted to prove that circ_0001018 and CCT4 (the host gene of circ_0001018) mRNA primarily localized in the cytoplasm of PTC cell lines. By qRT-PCR analysis, the expression of circ_0001018 and SOX4 mRNA was found upregulated while the expression of miR-338-3p was found downregulated in PTC tissues and cells. circ_0001018 silence significantly inhibited the tumor growth in xenografted nude mice. A series of cytological experiments such as a Cell Counting Kit-8 (CCK-8) assay, a 5-ethynyl-2′-deoxyuridine (EdU) assay, cell cycle profiling, wound healing, a transwell assay, and cell apoptosis were conducted and showed that circ_0001018 and SOX4 promoted cell proliferation, migration, and invasion, inhibited cell apoptosis, and reduced the cell cycle arrest at the G1 phase in PTC cells. Compared with circ_0001018 and SOX4, miR-338-3p held the opposite function. The regulatory relationship between circ_0001018 and miR-338-3p, and between miR-338-3p and SOX4 mRNA, was validated using a luciferase reporter gene assay and/or RNA immunoprecipitation (RIP assay). Our findings showed that circ_0001018 acted as the tumor promoter via sponging miR-338-3p to elevate SOX4 expression level in PTC. Importantly, this novel circ_0001018/miR-338-3p/SOX4 axis has the potential to be considered as a therapy target for PTC., Graphical Abstract, In present study, the authors’ findings showed that circ_0001018 acted as the tumor promoter via sponging miR-338-3p to elevate SOX4 expression level in papillary thyroid cancer (PTC). Importantly, this novel circ_0001018/miR-338-3p/SOX4 axis has the potential to be considered as a therapy target for PTC.

Differentiation of preadipocytes into functional adipocytes could be a major target for repressing obesity-induced insulin resistance (IR). However, the molecular mechanisms involved in adipogenesis and the development of IR are unclear. We report, for the first time, that miR-574-5p, a novel miRNA, promotes adipogenesis to suppress IR. An increase in the level of miR-574-5p significantly induced the differentiation of preadipocytes into mature adipocytes. Conversely, reduction of miR-574-5p levels blocked the differentiation of preadipocytes in vitro. In a dual-luciferase reporter assay, it was shown that homeobox A5 (HOXA5) promoted the transcription of miR-574-5p to induce the differentiation of preadipocytes. Hdac9, a direct downstream target of miR-574-5p, was involved in the regulation of adipocyte differentiation. The overexpression of miR-574-5p also promoted adipogenesis in subcutaneous fat to alleviate IR in high-fat-diet-fed mice. Additionally, miR-574-5p expression was significantly higher in the subcutaneous adipose tissue of obese patients without type 2 diabetes than in those with type 2 diabetes. There was an increase in HOXA5 expression and a decrease in histone deacetylase 9 (HDAC9) expression in the subcutaneous fat of obese patients without type 2 diabetes. These results suggest that miR-574-5p may be a potential therapeutic target for combating obesity-related IR., Graphical abstract, This is the first study to report that the upregulation of the expression of HOXA5-miR-574-5p-HDAC9 axis components can promote adipogenesis and maintenance of subcutaneous adipose expansion to suppress insulin resistance; this could be a potential therapeutic target for combating obesity-related insulin resistance.

Context: A solution of simethicone administrated orally before the scan endoscopy procedure is recommended as a good means to reduce the bubbles interference. Aims: To evaluate the reliability, reproducibility and effectiveness of simethicone pre-treatment before endoscopic exploration of patients not diagnosed with malignant digestive pathologies. Methods: The study was organized in two stages. Time zero was a retrospective reanalysis of data from 58 randomized patients (age 15-90 years) who had been evaluated by endoscopy without simethicone (Control group), and 66 patients (Simethicone group) who were recruited and received 62 mL of a simethicone pre-preparation drink, 30 min before examination. The second stage, using procedure with simethicone, was continued and after 6 months at the beginning, 88 subjects were randomized. Stability in the results was demonstrated with simethicone. Statistical analysis was performed using SPSS v.21. Results: Analysis of comorbidities were represented with the highest prevalence being high blood pressure, diabetes mellitus, long-term depression, obesity and hypothyroidism. Results demonstrated similarity among endoscopist and between analysts within the same group (examination time, images resolved and with the best quality, by mucosal areas). However, these variables were different between groups, such that valuation quality was improved with simethicone, even 6 months after the initial evaluation. Conclusions: Use of simethicone as a pre-preparation drink 30 min before the endoscopy evaluation improves the quality of images and reduces the time of operation.

Cancer remains one of the leading causes of death worldwide. Cancer stem cells (CSCs) are the underlying reason for tumor recurrence, progression, and therapeutic resistance. Aptamers are synthetic single-stranded oligonucleotides that can specifically bind to various molecular targets. Here, we aim to develop an effective aptamer-based biomarker and therapeutic tool that targets CSCs for cancer therapy. We perform whole-cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX) to screen DNA aptamers that specifically bound to lung CSCs, modeled by E-cadherin-silenced A549 cells. We develop a CSC-specific aptamer (AP-9R) specifically recognizing lung CSCs with high affinity and identify Annexin A2, a Ca2+-dependent membrane-binding protein, as its target. Annexin A2 expression was upregulated in lung CSCs and involved in cancer stemness. The expression of Annexin A2 was associated with signatures of stemness and metastasis, as well as poor clinical outcomes, in lung cancer in silico. Moreover, AP-9R decreased Annexin A2 expression and suppressed CSC properties in CSCs in vitro and in vivo. The present findings suggest that Annexin A2 is a CSC marker and regulator, and the CSC-specific aptamer AP-9R has potential theranostic applications for lung cancer.

Precise genome editing of human pluripotent stem cells (hPSCs) is crucial not only for basic science but also for biomedical applications such as ex vivo stem cell therapy and genetic disease modeling. However, hPSCs have unique cellular properties compared to somatic cells. For instance, hPSCs are extremely susceptible to DNA damage, and therefore Cas9-mediated DNA double-strand breaks (DSB) induce p53-dependent cell death, resulting in low Cas9 editing efficiency. Unlike Cas9 nucleases, base editors including cytosine base editor (CBE) and adenine base editor (ABE) can efficiently substitute single nucleotides without generating DSBs at target sites. Here, we found that the editing efficiency of CBE was significantly lower than that of ABE in human embryonic stem cells (hESCs), which are associated with high expression of DNA glycosylases, the key component of the base excision repair pathway. Sequential depletion of DNA glycosylases revealed that high expression of uracil DNA glycosylase (UNG) not only resulted in low editing efficiency but also affected CBE product purity (i.e., C to T) in hESCs. Therefore, additional suppression of UNG via transient knockdown would also improve C to T base substitutions in hESCs. These data suggest that the unique cellular characteristics of hPSCs could determine the efficiency of precise genome editing., Graphical abstract, Park and colleagues found that editing efficiency of CBE is lower than that of ABE in hPSCs, where expressions of DNA glycosylases are higher compared to the differentiated somatic counterparts. High UNG expression in hPSCs significantly affects CBE efficiency and purity. Thus, transient depletion of UNG would be beneficial for improvement of CBE efficiency in hPSCs.

In vitro transcribed (IVT)-mRNA has been accepted as a promising therapeutic modality. Advances in facile and rapid production technologies make IVT-mRNA an appealing alternative to protein- or virus-based medicines. Robust expression levels, lack of genotoxicity, and their manageable immunogenicity benefit its clinical applicability. We postulated that innate immune responses of therapeutically relevant human cells can be tailored or abrogated by combinations of 5′-end and internal IVT-mRNA modifications. Using primary human macrophages as targets, our data show the particular importance of uridine modifications for IVT-mRNA performance. Among five nucleotide modification schemes tested, 5-methoxy-uridine outperformed other modifications up to 4-fold increased transgene expression, triggering moderate proinflammatory and non-detectable antiviral responses. Macrophage responses against IVT-mRNAs exhibiting high immunogenicity (e.g., pseudouridine) could be minimized upon HPLC purification. Conversely, 5′-end modifications had only modest effects on mRNA expression and immune responses. Our results revealed how the uptake of chemically modified IVT-mRNA impacts human macrophages, responding with distinct patterns of innate immune responses concomitant with increased transient transgene expression. We anticipate our findings are instrumental to predictively address specific cell responses required for a wide range of therapeutic applications from eliciting controlled immunogenicity in mRNA vaccines to, e.g., completely abrogating cell activation in protein replacement therapies.

Antisense gapmer oligonucleotides containing phosphoryl guanidine (PG) groups, e.g., 1,3-dimethylimidazolidin-2-imine, at three to five internucleotidic positions adjacent to the 3′ and 5′ ends were prepared via the Staudinger chemistry, which is compatible with conditions of standard automated solid-phase phosphoramidite synthesis for phosphodiester and, notably, phosphorothioate linkages, and allows one to design a variety of gapmeric structures with alternating linkages, and deoxyribose or 2′-O-methylribose backbone. PG modifications increased nuclease resistance in serum-containing medium for more than 21 days. Replacing two internucleotidic phosphates by PG groups in phosphorothioate-modified oligonucleotides did not decrease their cellular uptake in the absence of lipid carriers. Increasing the number of PG groups from two to seven per oligonucleotide reduced their ability to enter the cells in the carrier-free mode. Cationic liposomes provided similar delivery efficiency of both partially PG-modified and unmodified oligonucleotides. PG-gapmers were designed containing three to four PG groups at both wings and a central “window” of seven deoxynucleotides with either phosphodiester or phosphorothioate linkages targeted to MDR1 mRNA providing multiple drug resistance of tumor cells. Gapmers efficiently silenced MDR1 mRNA and restored the sensitivity of tumor cells to chemotherapeutics. Thus, PG-gapmers can be considered as novel, promising types of antisense oligonucleotides for targeting biologically relevant RNAs., Graphical abstract, Many successful antisense oligonucleotide designs are gapmers, which contain a central section of DNA phosphorothioates flanked by RNA-like nucleotides. We describe a new type of gapmers with phosphoryl guanidine groups in the flanks, which efficiently silenced MDR1 gene expression and restored the sensitivity of drug-resistant tumor cells to conventional chemotherapy.

Context: Diabetes mellitus is a condition, which is characterized by persistent hyperglycemia, abnormal functioning of insulin and difficulty in the metabolism of carbohydrates, fats and proteins. Antidiabetic medicines of herbal origin are widely consumed in Pakistan. Therefore, there was a need to generate data on antidiabetic herbal formulations manufactured and marketed in Pakistan. Aims: To develop a list of Pakistani manufactured antidiabetic herbal products and the most common herbal ingredients found in them. Methods: Antidiabetic herbal formulations were collected from the renowned herbal stores of Karachi, Pakistan, and their ingredients were checked. The most common herbal ingredients found in them were determined, and the possible constituents responsible for the antidiabetic action of these herbs were discovered. Results: A total of 15 herbal antidiabetic products were collected from the herbal stores. The most common herbs found in them include Syzygium cumini, Gymnemma sylvestre, Aloe vera, Nigella sativa, Acacia nilotica, Commiphora myrrha, Portulaca oleracea, Punica granatum, Rhus coriaria, Coriandrum sativum, Trigonella foenum-graecum, Bambusa bambos, Holarrhena antidysenterica, Swertia changii, Curcuma longa and Rumex vesicarius. The constituents responsible for antidiabetic activity include alkaloids, polysaccharides, glycosides, secoiridoid, phenolic acids, flavonoids, terpenoids, saponins and volatile oil. Conclusions: The herbs incorporated in these formulations have proven antidiabetic effects; therefore, these formulations can produce significant results in the management of diabetes. More research on each of these formulations is needed to confirm the potency of all these herbal products.

Activation of hepatic stellate cells (HSCs) causes hepatic fibrosis and results in chronic liver diseases. Although activated HSC functions are facilitated by an increase in the cytosolic Ca2+ concentration ([Ca2+]cyt), the pathophysiological roles of ion channels are largely unknown. In the present study, functional analyses of the two-pore domain K+ (K2P) channels, which regulate the resting membrane potential and [Ca2+]cyt, were performed using the human HSC line, LX-2. Expression analyses revealed that TREK1 (also known as KCNK2 and K2P2.1) channels are expressed in LX-2 cells. Whole-cell K+ currents were activated by 10 μM arachidonic acid and the activation was abolished by 100 μM tetrapentylammonium, which are pharmacological characteristics of TREK1 channels. The siRNA knockdown of TREK1 channels caused membrane depolarization and reduced [Ca2+]cyt. In addition, TREK1 knockdown downregulated the gene expression of collage type I and platelet-derived growth factor. Furthermore, TREK1 knockdown inhibited the proliferation of LX-2 cells. In conclusion, the activity of TREK1 channels determines the resting membrane potential and [Ca2+]cyt, which play a role in extracellular matrix production and cell proliferation in HSCs. This study may help elucidate the molecular mechanism underlying hepatic fibrosis in HSCs and provide a potential therapeutic target for hepatic fibrosis.

Two major posttranscriptional mechanisms—alternative splicing (AS) and alternative polyadenylation (APA)—have attracted much attention in cancer research. Nevertheless, their roles in clear cell renal carcinoma (ccRCC) are still ill defined. Herein, this study was conducted to uncover the implications of AS and APA events in ccRCC progression. Through consensus molecular clustering analysis, two AS or APA RNA processing phenotypes were separately constructed with distinct prognosis, tumor-infiltrating immune cells, responses to immunotherapy, and chemotherapy. The AS or APA score was constructed to quantify AS or APA RNA processing patterns of individual ccRCCs with principal-component analysis. Both high AS and APA scores were characterized by undesirable survival outcomes, relatively high response to immunotherapy, and low sensitivity to targeted drugs, such as sorafenib and pazopanib. Moreover, several small molecular compounds were predicted for patients with a high AS or APA score. There was a positive correlation between AS and APA scores. Their interplay contributed to poor prognosis and reshaped the tumor immune microenvironment. Collectively, this study is the first to comprehensively analyze two major posttranscriptional events in ccRCC. Our findings uncovered the potential functions of AS and APA events and identified their therapeutic potential in immunotherapy and targeted therapy.

Context: Hyperglycemia causes diabetes mellitus (DM), abnormal metabolism, oxidative stress, and chronic complications such as impotence. Hyperglycemia causes testicular atrophy and stromal cell, seminiferous tubular damage, and spermatogenic cells. Bischofia javanica Blume is a plant that is used for the treatment of various chronic conditions and has traditionally by the people of Indonesia as a diabetes medicine. Aims: To determine the effect of B. javanica extract on the increase in the quality and quantity of sperm of hyperglycemic rats. Methods: The treatment groups consisted of; G0: negative control (-), G1: positive control (DM: alloxan induction + standard feed), G2-G4: DM + 300, 600 and 900 mg/kg BW of B. javanica leaves ethanol extract, respectively, and G5: DM + glibenclamide 0.5 mg/kg BW. Rats were dissected, and then the testes were taken to analyze sperm quantity and quality and immunohistochemistry. Results: There was a significant difference (p

Extracellular signal-regulated protein kinase 5 (ERK5) has various physiological functions. However, the physiological role of ERK5 in the treatment of mice with an illicit drug such as methamphetamine (METH) remains unknown. We revealed that mice treated with METH showed hyperactivity, and increased p-ERK5 and Iba1 (a microglia marker) levels in the striatum. Additionally, these changes were inhibited by pretreatment with the ERK5 inhibitor BIX02189. The results suggest that METH-induced hyperactivity is associated with the activation of microglia via p-ERK5 in the striatum. Thus, the ERK5 pathway components in the central nervous system are potential therapeutic targets for preventing METH addiction.

Gene editing utilizing homology-directed repair has advanced significantly for many monogenic diseases of the hematopoietic system in recent years but has also been hindered by decreases between in vitro and in vivo gene integration rates. Homology-directed repair occurs primarily in the S/G2 phases of the cell cycle, whereas long-term engrafting hematopoietic stem cells are typically quiescent. Alternative methods for a targeted integration have been proposed including homology-independent targeted integration and precise integration into target chromosome, which utilize non-homologous end joining and microhomology-mediated end joining, respectively. Non-homologous end joining occurs throughout the cell cycle, while microhomology-mediated end joining occurs predominantly in the S phase. We compared these pathways for the integration of a corrective DNA cassette at the Bruton’s tyrosine kinase gene for the treatment of X-linked agammaglobulinemia. Homology-directed repair generated the most integration in K562 cells; however, synchronizing cells into G1 resulted in the highest integration rates with homology-independent targeted integration. Only homology-directed repair produced seamless junctions, making it optimal for targets where insertions and deletions are impermissible. Bulk CD34+ cells were best edited by homology-directed repair and precise integration into the target chromosome, while sorted hematopoietic stem cells contained similar integration rates using all corrective donors., Graphical Abstract, There are alternative methods for nuclease-mediated targeted gene integration, including homology-independent targeted integration (HITI) and precise integration into target chromosome (PITCh), which utilize non-homologous end joining and microhomology-mediated end joining, respectively. These DNA repair pathways can be utilized in both cell lines and primary hematopoietic stem cells.

Rheumatoid arthritis (RA) is an inflammation-involved disorder and features the disruption of CD4+ T lymphocytes. Herein, we describe that microRNA-10b-5p (miR-10b) promotes RA progression by disrupting the balance between subsets of CD4+ T cells. MiR-10b-deficient mice protected against collagen antibody-induced arthritis (CAIA) model. RNA sequencing results indicated that disordered genes in miR-10b−/− CAIA model are closely associated with CD4+ T cells differentiation. Moreover, miR-10b mimics promoted Th1/Th17 and suppressed Th2/Treg cells differentiation, whereas miR-10b inhibitor induced contrary effects. In addition, GATA3 and PTEN was confirmed as two targets of miR-10b, and GATA3 siRNA could increase Th1 and reduce Th2 cells meanwhile PTEN siRNA could increase Th17 and decrease Treg cells. Furthermore, miR-10b inhibitor significantly ameliorated collagen-induced arthritis (CIA) development by attenuating the dysfunctional CD4+ T cell subpopulations. The present findings suggest that miR-10b could disrupt the balance of CD4+ T subsets, while suppressed miR-10b could attenuate the severity of experimental arthritis, which provided us a novel mechanistic and therapeutic insight into the RA.

Glaucoma is the leading cause of irreversible vision loss worldwide, and reduction of intraocular pressure (IOP) is the only factor that can be interfered to delay disease progression. As the first line and preferred method to treat glaucoma, eye drops have many shortcomings, such as low bioavailability, poor patient compliance, and unsustainable therapeutic effect. In this study, a highly efficient brimonidine (BRI) silicone rubber implant (BRI@SR@TPU implant) has been designed, prepared, characterized, and administrated for sustained relief of IOP to treat glaucoma. The in vitro BRI release from BRI@SR@TPU implants shows a sustainable release profile for up to 35 d, with decreased burst release and increased immediate drug concentration. The carrier materials are not cytotoxic to human corneal epithelial cells and conjunctival epithelial cells, and show good biocompatibility, which can be safely administrated into rabbit’s conjunctival sac. The BRI@SR@TPU implant sustainably released BRI and effectively reduced IOP for 18 d (72 times) compared to the commercial BRI eye drops (6 h). The BRI@SR@TPU implant is thus a promising noninvasive platform product for long-term IOP-reducing in patients with glaucoma and ocular hypertension.

Acipimox, a nicotinic acid derivative in clinical use for the treatment of hyperlipidaemia, incorporates a free carboxylic acid and an N-oxide moiety, functionalities known to interact with the metalloenzyme carbonic anhydrase (CA, EC 4.2.1.1) and inhibit its activity. Herein we report that acipimox acts as a low micromolar CA inhibitor (CAI) against most human (h) isoforms possessing catalytic activity, hCA I - XIV. By using computational techniques (docking and molecular dynamics simulations), we propose that acipimox coordinates through its carboxylate group to the zinc ion from the enzyme active site cavity, whereas the N-oxide group is hydrogen-bonded to the proton shuttle His residue in some isoforms (hCA I) or to active site Thr or Gln residues in other isoforms (hCA II, III, IV, VII, etc). As some CA isoforms are involved in lipogenesis, these data may be useful for the design of more effective CAIs with antiobesity activity.

Context Delphinidin-3-O-glucoside (DP) is a bioactive compound of Hibiscus sabdariffa L. (Malvaceae) (Roselle) calyces and exerts endothelial protection and lipid-lowering activities, which provided a basis for the prevention and treatment of cardiovascular diseases. Objectives To investigate the therapeutic effects of DP against atherosclerosis. Materials and methods A rabbit model of atherosclerosis (AS) was established by 12 weeks of a high-fat diet (HFD). The rabbits were divided into five groups: control, AS, simvastatin (4 mg/kg), and two DP groups (10 and 20 mg/kg). After treatment with DP or simvastatin by oral gavage for 12 weeks, the lipid profiles were measured. Histopathological assessment of the aorta was performed by H&E staining. Oxidative stress and inflammation-related markers were analyzed by ELISA kit and real-time RT-PCR. Results DP (20 mg/kg) decreased serum TG (2.36 ± 0.66 vs. 4.33 ± 0.27 mmol/L for the AS group), TC, LDL-C, and HDL-C (all p

Rheumatoid arthritis (RA) is an ordinarily occurring autoimmune disease with systemic inflammatory. Targeted drug delivery systems have many successful applications in the treatment of rheumatoid arthritis. In order to develop nanoparticles for targeted delivery of Celastrol (Cel) to rheumatoid arthritis and specific drug release, the dextran sulfate (DS) was modified as the targeting molecular by binding to the scavenger receptor of macrophage. The dextran-sulfate-PVGLIG-celastrol (DS-PVGLIG-Cel), named DPC, amphiphilic polymeric prodrug was synthesized and characterized. The resulting DPC@Cel micelles had the average size of 189.9 nm. Moreover, the micelles had ultrahigh entrapment efficiency (about 44.04%) and zeta potential of −11.91 mV. In the in vitro release study, due to the excessive production of matrix metalloproteinase-2 (MMP-2) at the inflammatory joint, the MMP-2 reactive peptide was used to crack in the inflammatory microenvironment to accelerate the release of Cel. The results have shown that the nanoparticles can effectively deliver Cel to activated macrophages and significantly improve the bioavailability. In vivo experiments showed that DPC@Cel have better anti-rheumatoid arthritis effects and lower systemic toxicity than free Cel. This study provided a new therapeutic strategy for the treatment of RA.

The screened compound DYT-1 from our in-house library was taken as a lead (inhibiting tubulin polymerisation: IC50=25.6 µM, anti-angiogenesis in Zebrafish: IC50=38.4 µM, anti-proliferation against K562 and Jurkat: IC50=6.2 and 7.9 µM, respectively). Further investigation of medicinal chemistry conditions yielded compound 29e (inhibiting tubulin polymerisation: IC50=4.8 µM and anti-angiogenesis in Zebrafish: IC50=3.6 µM) based on tubulin and zebrafish assays, which displayed noteworthily nanomolar potency against a variety of leukaemia cell lines (IC50= 0.09–1.22 µM), especially K562 cells where apoptosis was induced. Molecular docking, molecular dynamics (MD) simulation, radioligand binding assay and cellular microtubule networks disruption results showed that 29e stably binds to the tubulin colchicine site. 29e significantly inhibited HUVEC tube formation, migration and invasion in vitro. Anti-angiogenesis in vivo was confirmed by zebrafish xenograft. 29e also prominently blocked K562 cell proliferation and metastasis in blood vessels and surrounding tissues of the zebrafish xenograft model. Together with promising physicochemical property and metabolic stability, 29e could be considered an effective anti-angiogenesis and -leukaemia drug candidate that binds to the tubulin colchicine site.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was identified as the cause of coronavirus disease 20019 (COVID19) pandemic which first emerged in December 2019 in Wuhan city, China. Currently, a vaccine is urgently needed to control the COVID-19 pandemic. Several vaccine candidates are under development and some are in the final stage of clinical trials. The COVID-19 vaccination aims to reduce morbidity and mortality rates, achieve herd immunity to prevent and protect the society, strengthen the health system, maintain productivity and minimize social and economic impacts. Before approval, vaccines have to undergo several clinical trials to ensure its safety profile, efficacy, duration of immune system resistance, and adverse effect. Various strategies have been used in the development of vaccines including viral vector vaccines, nucleic acid vaccines, inactivated virus, live attenuated virus, subunit protein¸and virus-like particle vaccine. Each strategy has its own advantages and disadvantages.

Background. Failure of dentures that are unable to withstand flexural forces is often a major problem in the use of metal-ceramic crowns. The use of rice husk nano-silica powder has been extensively researched in dentistry because of its ability to improve mechanical properties. This study aims to determine the effectiveness of nano-silica particles from rice husks on the flexural strength of the opaque layer on Co-Cr metal-ceramic crowns. Methods. The type of research is experimental laboratories in vitro with posttest only control group design. The research samples were rectangular Co-Cr metal and porcelain plated on top of the metal center totaling 25 samples which were divided into 5 treatment groups. Flexural strength measurement with three-point bending at UTM. Data were analyzed by independent T-test and way ANOVA. Results. flexural strength of Co-Cr metal ceramic crown with the addition of 0.25% rice husk silica; 0.5%; 0.75%.1% in the opaque layer is 70.431±4.168; 84.093±2.852; 100,672±4,182 and 115,092±3,821, respectively. Conclusion. There is an increase in the flexural strength of metal ceramic crowns with the addition of 0.25% silica; 0.5%; 0.75%,1% on opaque coating.

Background: Testosterone replacement therapy (TRT) is one of the main lines of treatment for men with hypogonadism. This study sought to evaluate the influence of TRT in men with late-onset hypogonadism (LOH), regarding fatigue, coronary artery disease (CAD), carotid intima-media thickness (CIMT) and cardiovascular risk. Methods: This study compared men with LOH already on TRT for >1 year to newly diagnosed men with LOH who recently started TRT (controls). We included men aged >18 years with clinical manifestations of testosterone deficiency and testosterone levels of 1 year and 45 (±12.2) years in the control group (p=0.18). CAD was present in 14 (46.7%) patients in the control group and in 3 (9.1%) in the study group (p1 year was not associated with lower rates of CAD in multivariable analysis. Fatigue Severity Score was significantly higher in the control group (39.2±15.0), compared to TRT >1 year (23.5±8.1; p1 year was associated with a 14.8-point decrease in Fatigue Severity Score (p1 year was associated with significantly lower fatigue scores. No differences were observed regarding CIMT, CAD and cardiovascular risk according to the WHO-ISH scale.

Mechanical stimulation of cultured keratinocytes and a living epidermis increases intracellular calcium ion concentrations ([Ca2+]i) in stimulated cells. This action propagates a Ca2+ wave to neighboring keratinocytes via ATP/P2Y2 receptors. Recent behavioral, pharmacological studies revealed that exogenous ATP induces itching via P2X3 receptors in mice. We previously showed that alloknesis occurs when an external stimulus is applied to the skin with increased epidermal histamine in the absence of spontaneous pruritus. Based on these results, we investigated the effects of histamine at a concentration that does not cause itching on ATP-induced itching. The mean number of scratching events induced by the mixture of ATP and histamine increased by 28% over the sum of that induced by histamine alone or ATP alone. A317491, a P2X3 receptor antagonist, suppressed the mixture-induced scratching more often than the ATP-induced scratching. Next, we examined the ATP-induced [Ca2+]i change before and after histamine stimulation using normal human epidermal keratinocytes. Some cells did not respond to ATP before histamine stimulation but responded to ATP afterward, the phenomenon suppressed by chlorpheniramine maleate. These findings suggest that histamine enhances ATP-induced itching and that a potential mechanism could involve increased responsiveness to ATP in keratinocytes.

Megalin receptor-mediated endocytosis participates a crucial role in gentamicin (GM) uptake, accumulation, and toxicity. In this study, we investigated the potential effects of montelukast (MLK) on megalin expression/endocytic function against GM nephrotoxicity. Male Wistar rats were administered GM (120 mg/kg; i.p.) daily in divided doses along 4 hr; 30 mg/kg/hr; for 7 days. MLK (30 mg/kg/day) was orally administered 7 days before and then concurrently with GM. The protein expressions of megalin and chloride channel-5 (ClC-5); one of the essential regulators of megalin endocytic function; were determined by Western blotting. Besides, the endocytic function of megalin was evaluated by the uptake of bovine serum albumin labeled with fluorescein isothiocyanate (FITC-BSA) into proximal tubular epithelial cells. Moreover, kidney function biomarkers (Cr, BUN, GFR, KIM-1, cystatin-C) and apoptosis markers (p-AKT1, cleaved caspase-3) were estimated. Co-treatment with MLK downregulated ClC-5 expression leading to reduced recycling of megalin to the plasma membrane, reduced expression, and so impaired endocytic function that was evidenced by reduced uptake of FITC-BSA in proximal tubular epithelial cells. The protein expression of the apoptotic executioner cleaved caspase-3 was significantly reduced, while that of the antiapoptotic p-AKT1 was elevated. These results were confirmed by the improvement of kidney functions and histological findings. Our data suggest that MLK could interfere with megalin expression/endocytic function that could be attributed to downregulation of ClC-5 protein expression. That eventually reduces renal cell apoptosis and improves kidney functions after GM administration without affecting the antibacterial activity of GM. Therefore, reduced expression of ClC-5 and interference with megalin expression/endocytic function by MLK could be an effective strategy against GM nephrotoxicity.

Red Sea represents one of the most remarkable marine ecosystems. However, it is also one of the world's least explored areas of marine biodiversity. The aims of this investigation were therefore, to isolate marine microorganisms from the seashore sediments and water in shallow region from west Yemen coast, to assess their antimicrobial potential, to identify the highly active isolate, and to purify and identify the bioactive compounds from it. In this regard, twenty-five bacterial strains have been isolated from twenty samples and tested for their antimicrobial ability against some pathogenic bacteria and yeast by using the agar disk diffusion and agar well diffusion assay. Out of the total 25 marine actinomycetes isolates only 13 exhibited interesting antimicrobial activity. The morphological, biochemical, and phylogenetic characteristics of the potential isolate 1S1 were compatible with their classification in the genus Streptomyces. The 16S rRNA gene sequences have shown that the isolate 1S1 clustered with Streptomyces longisporoflavus. The strain Streptomyces sp. 1S1 was cultivated and extracted with ethyl acetate. The GC–MS study of the extract indicated the presence of certain fatty acyl compounds e.g., tetradecanoic acid, 9-octadecenoic acid, hexadecanoic acid, and 9,12,15-octadecatrienoic acid. Using chromatographic techniques, three compounds were isolated and by spectroscopic methods e.g., IR, MS and NMR structurally elucidated. The three compounds were identified as a triacylglyceride, 9-octadecenoic acid, and hexadecanoic acid. The study reinforces the evidence of the potential of Streptomyces sp and the ability to produce several antimicrobial compounds.

Background. Endometriosis is a chronic gynecological disorder that generally affects women of reproductive age which is characterized by implantation of endometrial tissue, both glands and stroma outside the uterus. Research shows that the initial onset of endometriosis patients begins in adolescence. The high prevalence with atypical symptoms leads to long term delay in establishing the appropriate diagnosis. Further developments in the field of early endometriosis screening will greatly assist clinicians, especially general practitioners in areas with limited health facilities, in early detection of endometriosis. Methods. Using the electronic databases, comprehensive literature searches were conducted with the specific keywords. Full-text manuscripts published were reviewed for relevancy and importantly, reference lists were cross-checked for additional relevant studies. Results. Dysmenorrhea, chronic pelvic pain, dyspareunia, dysuria, dyschezia, and glycoprotein were found dominantly in all analyzed literature. The occurrence of endometriosis affects the functioning of women in multiple aspects of life. Endometriosis places a significant burden on teens and adult women, their families, and society as a whole. Conclusion. Combination of structured past history taking, and several biomarker level might be useful as a screening tool that easy to use and cost effective, as early diagnosis is expected to prevent further progression of endometriosis especially in adolescence.

Liver fibrosis is a common pathologic stage of the development of liver failure. It has showed that exosomes loaded with therapeutic circRNAs can be manufactured in bulk by exosome secreted cells in vitro, thus enabling personalized treatment. This study aimed to investigate the role of exosome-based delivery of circDIDO1 in liver fibrosis. Levels of genes and proteins were examined by qRT-PCR and Western blot. Cell proliferation, apoptosis, and cell cycle were analyzed by using cell counting kit-8 (CCK-8) assay, EdU assay, and flow cytometry, respectively. The binding between circDIDO1 and miR-141-3p was confirmed by dual-luciferase reporter, RNA pull-down and RIP assays. Exosomes were isolated by ultracentrifugation, and qualified by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blot. CircDIDO1 overexpression or miR-141-3p inhibition suppressed the proliferation, reduced pro-fibrotic markers, and induced apoptosis as well as cell cycle arrest in hepatic stellate cells (HSCs) by blocking PTEN/AKT pathway. Mechanistically, circDIDO1 acted as an endogenous sponge for miR-141-3p, further rescue experiments showed that circDIDO1 suppressed HSC activation by targeting miR-141-3p. Extracellular circDIDO1 could be incorporated into exosomes isolated from mesenchymal stem cells (MSCs), and transmitted to HSCs to restrain HSC activation. Clinically, low levels of serum circDIDO1 in exosome were correlated with liver failure, and serum exosomal circDIDO1 had a well diagnostic value for liver fibrosis in liver failure patients. Transfer of circDIDO1 mediated by MSC-isolated exosomes suppressed HSC activation through the miR-141-3p/PTEN/AKT pathway, gaining a new insight into the prevention of liver fibrosis in liver failure patients.

Backgrounds. It is very important to evaluate and assess the rationality of the use of antidiabetic drugs, especially the biguanide (metformin) group to maintain the quality and quality of diabetes mellitus drug administration so that the target of diabetes mellitus control can be optimized. This study aims to evaluate the rationality of prescribing metformin oral antidiabetic at Dr. Mohammad Hoesin General Hospital Palembang. Methods. The research design is a descriptive study using secondary data from medical records in the medical records section of Dr. Moh Hoesin Hospital Palembang. The object of research is all medical records of patients with type 2 diabetes mellitus who used metformin at RSMH Palembang in the period July 1st, 2019-July 31st, 2020, with complete medical record data and without serious comorbidities. The rationality for using metformin that was assessed in this study was the frequency of use, drug dose, route of administration, duration of administration, and drug interactions. The frequency of drug use is assessed by how many times the drug is taken in one day. Results. The most age group of patients who received a prescription for metformin were 51-60 years old and 61-70 years old with a total of 17 patients (35.4%). The majority of patients were female as much as 60.4%. Drug interactions with metformin are still quite common, although the majority are synergistic and potentiating interactions. There are still 2 cases or 4.2 percent who experience antagonistic interactions. Conclusion. The rationality for using metformin in patients with type 2 diabetes mellitus is based on the criteria for the right dose (100%), the right frequency of drug administration (100%), the right time for giving the drug (100%), the right way of giving the drug (100%), and the right drug interaction. (95.8%).

Background. Bronchiectasis is a chronic inflammatory airway disease characterized by chronic bronchial dilatation. The cause of bronchiectasis is still ideopathic but most of the bronchiectasis are caused by tuberculosis infection and will interfere with the patient's quality of life. In addition, bronchiectasis can be caused by aspiration of foreign bodies. Case presentation. A 42-year-old male patient was treated at Dr, M Djamil Padang Hospital with complaints of coughing up blood, in addition the patient complained of coughing up yellowish phlegm, shortness of breath and fever. The patient had a history of foreign body aspiration 6 months ago. The patient had a chest X-ray and a chest CT scan and a picture of bronchiectasis and mulltiple bullae was found. The patient also had pulmonary tuberculosis infection where the X-pert gene examination in the patient found that MTB was not detected. Conclusion. The patient had multiple bullae, which were complications of emphysema, and the patient had or had compensatory emphysema, where the abnormality was hyperinflation of certain parts of the lung due to atelectasis. The condition of bronchiectasis and multiple bullae makes the patient susceptible to infection, which in this case was found to have pulmonary tuberculosis infection. Management of bronchiectasis includes identification of acute exacerbations and the use of antibiotics, controlling microbial growth, treatment of the underlying condition, and reducing the excessive inflammatory response.