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Background Chronic limb-threatening ischemia (CLTI) is the most advanced stage of peripheral artery disease (PAD) and has poor clinical outcomes. Recently, stimulating arteriogenesis has been proposed to improve clinical outcomes. Several studies have shown that miRNAs have beneficial effects on limb ischemia related to arteriogenesis. This study aimed to review the roles of therapeutic miRNAs in the arteriogenesis of limb ischemia. Methods A systematic search was conducted through July 2021 using the PubMed, Scopus, and ScienceDirect databases. Two authors independently assessed studies that investigated the role of miRNAs in the arteriogenesis of limb ischemia, both in vivo and in clinical studies. Results All selected studies were in vivo studies, with a total of 36 articles and 28 types of miRNAs. miRNAs potentially regulate arteriogenesis by targeting different targets. The following miRNAs were upregulated to enhance arteriogenesis: miRNA-126-3p, -93, -675, -143-3p, -130a, -210, -146b, -21, -let-7g, -132/212, -150, and 155. Meanwhile, microRNAs needed to be downregulated, namely: miRNA-939-5p, -503, -199a-5p, -146a, -92a, -14q32 microRNA gene cluster, -15a/16, -100, -133a, -139-5p, -223, -352, -615-5p, -15b/5p, -124-3p, and 29a. MiRNA-126 was the most studied miRNA, and SPRED1 was the most common target of microRNA. However, the included studies showed high heterogeneity in terms of inducing hindlimb ischemia, the timing of administration, and the method used for evaluating arteriogenesis. Moreover, most studies presented unclear or high-risk bias. Conclusion MicroRNA application in a preclinical model of hindlimb ischemia has beneficial effects on arteriogenesis. This result indicates that miRNAs might be potentially beneficial in patients with CLTI. Registration The review protocol was registered with PROSPERO under registration number CRD42024484988. more...

Abstract Chronic thromboembolic pulmonary hypertension (CTEPH) is marked by persistent blood clots in pulmonary arteries, leading to significant morbidity and mortality. Emerging evidence highlights the role of microRNAs (miRNAs) in pulmonary hypertension, though findings on miRNA expression in CTEPH remain limited and inconsistent. This systematic review evaluates miRNA expression changes in CTEPH and their direction. Following Preferred Reporting Items for Systematic Reviews and Meta‐Analyses guidelines, we registered our protocol in International Prospective Register of Systematic Reviews (CRD42024524469). We included studies on miRNA expression in CTEPH with comparative or analytical designs, excluding nonhuman studies, interventions, non‐English texts, conference abstracts, and editorials. Databases searched included PubMed, EMBASE, Scopus, CENTRAL, and ProQuest. The Quality Assessment of Diagnostic Accuracy Studies‐2 tool assessed bias risk, and results were synthesized narratively. Of 313 unique studies, 39 full texts were reviewed, and 9 met inclusion criteria, totaling 235 participants. Blood samples were analysed using quantitative real time polymerase chain reaction. Seven miRNAs (miR‐665, miR‐3202, miR‐382, miR‐127, miR‐664, miR‐376c, miR‐30) were uniformly upregulated, while nine (miR‐20a‐5p13, miR‐17‐5p, miR‐93‐5p, miR‐22, let‐7b, miR‐106b‐5p, miR‐3148, miR‐320‐a, miR‐320b) were downregulated in CTEPH patients. Two upregulated miRNAs (miR‐127 and miR‐30a) were consistently associated with previous evidence in the mechanism inducing the development of CTEPH, and five downregulated miRNAs (miR‐20‐a, miR‐17‐5p, miR‐93‐5p, let‐7b, miR‐106b‐5p) were associated with a protective effect against CTEPH. We also identified gaps in the literature where the evidence for five upregulated miRNAs (miR‐665, miR‐3202, miR‐382, miR‐664 and miR‐376c) and four downregulated miRNAs (miR‐22, miR‐3148, miR‐320‐a, and miR‐320b) in CTEPH is conflicting. Our findings offer insights into the role of miRNAs in CTEPH and underscore the need for further research to validate these miRNAs as biomarkers or therapeutic targets. more...

Background Chronic limb-threatening ischemia (CLTI) is the most advanced stage of peripheral artery disease (PAD) and has poor clinical outcomes. Recently, stimulating arteriogenesis has been proposed to improve clinical outcomes. Several studies have shown that miRNAs have beneficial effects on limb ischemia related to arteriogenesis. This study aimed to review the roles of therapeutic miRNAs in the arteriogenesis of limb ischemia. Methods A systematic search was conducted through July 2021 using the PubMed, Scopus, and ScienceDirect databases. Two authors independently assessed studies that investigated the role of miRNAs in the arteriogenesis of limb ischemia, both in vivo and in clinical studies. Results All selected studies were in vivo studies, with a total of 36 articles and 28 types of miRNAs. miRNAs potentially regulate arteriogenesis by targeting different targets. The following miRNAs were upregulated to enhance arteriogenesis: miRNA-126-3p, -93, -675, -143-3p, -130a, -210, -146b, -21, -let-7g, -132/212, -150, and 155. Meanwhile, microRNAs needed to be downregulated, namely: miRNA-939-5p, -503, -199a-5p, -146a, -92a, -14q32 microRNA gene cluster, -15a/16, -100, -133a, -139-5p, -223, -352, -615-5p, -15b/5p, -124-3p, and 29a. MiRNA-126 was the most studied miRNA, and SPRED1 was the most common target of microRNA. However, the included studies showed high heterogeneity in terms of inducing hindlimb ischemia, the timing of administration, and the method used for evaluating arteriogenesis. Moreover, most studies presented unclear or high-risk bias. Conclusion MicroRNA application in a preclinical model of hindlimb ischemia has beneficial effects on arteriogenesis. This result indicates that miRNAs might be potentially beneficial in patients with CLTI. Registration The review protocol was registered with PROSPERO under registration number CRD42024484988. more...

BACKGROUND: Ultraviolet B (UVB) induces skin photoaging by reducing collagen deposition via impairment of the TGF-β/Smad signaling pathway. Achatina fulica mucus (AFM) is a native medicine acting as vehicle of anti-aging ingredients. The present investigation examined the effect of AFM on UVB-induced fibroblast photoaging by assessing TGF-β, Smad3, and Smad7 mRNA expressions. METHODS: AFM was extracted from A. fulica using electrical shock and freeze-dried into a powder. Normal human dermal fibroblast (NHDF) cultures were irradiated with/without 100 mJ/cm2 UVB and treated with/without 10% platelet-rich plasma or different concentrations of AFM: 3.9 μg/mL in AF3 group; 15.625 μg/mL in AF15 group, and 62.5 μg/mL in AF62 group. The mRNA expressions of TGF-β, Smad3, and Smad7 in NHDF were evaluated by quantitative polymerase chain reaction. RESULTS: TGF-β mRNA expressions in the AF3 (0.85±0.01), AF15 (0.94±0.02) and AF62 (1.64±0.03) groups were significantly higher (p more...

Context: Acute myeloid leukemia (AML) is the most common form of acute leukemia. Currently, many people use medicinal herbs for the treatment of cancers. Nut grass (Cyperus rotundus L.) is a medicinal plant widely used in conventional medicine due to its role as an anti-cancer. Aims: To evaluate the effect of Cyperus rotundus tuber (CRT) ethanolic extract on cell proliferation, differentiation, cell cycle, and apoptotic of HL-60 cells. Methods: HL-60 cells line as a model for AML was cultured under the influence of CRT extract concentrations (35.4, 354, and 3540 µg/mL) for 48 h. Furthermore, the cell was subjected to carboxyfluorescein succinimidyl ester staining for proliferation, using a CD117 marker for differentiation. Annexin V-FITC and PI for cell cycle and apoptosis. The effect of this herb was studied by flow cytometry. The data were statistically analyzed with one-way ANOVA (p≤0.05) and the Tukey test using SPSS version 16 for Windows. Results: The results showed that the CRT inhibited proliferation activity, differentiation, cell cycle arrest, and apoptosis. Conclusions: The data clearly showed the potential anti-cancer activity of CRT on HL-60 cells and suggested that it could help develop promising therapeutic agents for AML treatments. more...

Background: Nasopharyngeal carcinoma (NPC) is endemic cancer in Southeast Asia with a relatively poor prognosis. Chemoradiotherapy is a primary treatment that advantages certain patients, particularly in the early stages. New predictive and prognostic biomarkers are required to guide and select the best treatment. Aims: To evaluate the circulation expression profile of microRNAs (miRNAs) associated with responses to chemoradiotherapy in nasopharyngeal carcinoma. Methods: Peripheral blood from 17 patients was collected before and after chemotherapy and radiotherapy. Differential expression circulating miRNAs were analyzed using microRNA Cancer Panels and were compared among patients with complete responses. Differential expression analysis using GenEx 7 Multid, statistic represented by GraphPad Prism 9. Alterations mechanism signaling pathways and biological function using IPA (Ingenuity Pathways Analysis). Results: Using microRNAs Cancer Plate consisting of 116 miRNAs, we identified ten circulating miRNAs that were differentially expressed in NPC patients after chemoradiotherapy. Unsupervised clustering and confirmation using qRT-PCR showed that miR-483-5p, miR-584-5p, miR-122-5p, miR-7-5p, miR-150-5p were overexpressed and miRNA are miR-421, miR-133a-3p, miR-18a-5p, miR-106b-3p, miR-339-5p were significantly downregulated after chemoradiotherapy (p more...

Background: Prostatic anomalies are common in tumor or infection condition. The enlargement of prostate gland affects the epithelial cell polarity that involves epithelial-mesenchymal transition (EMT). Transition into mesenchymal is mediated by transcription factor ZEB1 and E-cadherin protein. Upregulation of ZEB1 and loss of E-Cadherin expression were associated to proliferation and metastasis of malignancy cells. This study aims to describe the correlation of ZEB1 and E-cadherin expression in prostatic anomaly. Materials and method: Samples were Formalin Fixed Paraffin Embedded (FFPE) block consist of 8 block Benign Prostatic Hyperplasia (BPH), 6 blocks High Grade Prostatic Intraepithelial Neoplasia (HGPIN) and 6 blocks Prostate Carcinoma (PCA). The blocks then sliced into 5 sections to be prepared for RNA extraction procedures. ZEB1 and E-Cadherin expression was analyzed by semi-quantitative procedures using PCR and electrophoresis. Correlation between ZEB1 and E-Cadherin espression was analyzed using Spearman’s rank correlation. Results: Relative expression of ZEB1 and E-cadherin mRNA in each group of prostatic anomaly were not significantly different (p>0.05). ZEB1 and E-Cadherin mRNA expression showed a significant and moderate level of negative correlation (p more...

Abstract Objective This study aims to evaluate the correlation between electrolytes and serial miRNAs from our previous study. We want to prove that there is the molecular basis that underlying electrolytes disturbances as the predictive indicator to the outcome in NSCLC patients. Results There were positive correlation between potassium level with miR-34 (p = 0.008, r = 0.366), miR-148 (p = 0.004, r = 0.394) and miR-155 (p = 0.031, r = 0.300). more...

Latar belakang: Angka kekambuhan dan resistensi pasien kanker rektum mencapai 40 persen. Kondisi tersebut bisa disebabkan karena peningkatan ekspresi GLUT-1 dan VEGF-A, serta mempengaruhi prognosis pasien. Tujuan penelitian ini adalah untuk mengetahui korelasi ekspresi mRNA GLUT-1 dan VEGF-A, serta hubungannya dengan prognosis pasien kanker rektum yang menjalani kemoradiasi. Metode: Penelitian Kohor ini melibatkan 16 orang pasien kanker rektum lokal stadium II atau III yang menjalani kemoradiasi di RSUP Kariadi Semarang. Sampel darah intravena diambil 5 mL pada saat sebelum dan sesudah kemoradiasi. Total RNA diisolasi dari 200 μl serum, kemudian dilakukan sintesis cDNA. Ekspresi mRNA GLUT-1 dan VEGF-A dikuantifikasi dengan metode Livak menggunakan reference gene β-actin. Hasil: Rata-rata ekspresi mRNA GLUT-1 menurun signifikan 2,14 kali (P=0,044) dan mRNA VEGF-A menurun 1,9 kali (P = 0,03). Ekspresi mRNA GLUT-1 dan VEGF-A berkorelasi positif kuat dan signifikan pada saat sebelum (r = 0,6; R2 = 0,455; P = 0,013) dan sesudah kemoradiasi (r = 0,8; R2 = 0,598; P < 0,001). Peningkatan ekspresi mRNA GLUT-1 berhubungan dengan prognosis buruk pasien, dengan resiko 18 kali lebih tinggi (P=0,036; OR=18, 95% CI=1,2 – 261). Peningkatan ekspresi mRNA VEGF-A tidak berhubungan signifikan dengan prognosis pasien (P=0,12; OR=9; 95%CI=0,6-123). Kesimpulan: Ekspresi mRNA GLUT-1 dan VEGF-A berkorelasi positif dan saling mempengaruhi satu dengan lainnya. Peningkatan ekspresi mRNA GLUT-1 berhubungan dengan prognosis buruk pasien. Hasil penelitian ini mengindikasikan bahwa ekspresi mRNA GLUT-1 dari sampel darah berpotensi sebagai biomarker prognosis pada pasien kanker rektum yang menjalani kemoradiasi. Kata kunci: kanker rektum, kemoradiasi, GLUT-1, VEGF-A, prognosis Abstract Background: Rectal cancer patients have 40 percent risk of recurrence and resistance, which is triggered by increasing in GLUT-1 and VEGF-A mRNA expression. This condition associate with the patients prognosis. This study aimed to determine the correlation between GLUT-1 and VEGF-A mRNA expression, and analyze its association with the rectal cancer patients prognosis who received chemoradiation. Methods: This was a Cohort study involving 16 rectal cancer patients with stage II or III undergoing chemoradiation at Kariadi Hospital Semarang. Five milliliters of intravenous blood samples were taken before and after chemoradiation. Total RNA was isolated from 200 μl of blood serum, followed by cDNA synthesis. GLUT-1 and VEGF-A mRNA expression was quantified by the Livak method using β-actin as a reference gene. Results: GLUT-1 and VEGF-A mRNA expression decreased significantly 2.14 times (P=0,044) and 1,9 times (P=0,03), respectively. Expression of GLUT-1 and VEGF-A mRNA have a significant and strong positive correlation at before (r=0,6; R2=0,455; P=0,013) and after chemoradiation (r=0,8; R2=0,598; P more...

Latar Belakang: Gangguan regulasi mikroRNA(miR) dan inflamasi kronik dapat mengubah tumor menjadi karsinoma dan kanker dengan metastasis melalui perubahan seluler dan genomik. Lesi prekanker memiliki peluang 33,3 persen menjadi kanker. Penelitian ini bertujuan untuk mengkaji peran miR-155-5p terhadap mRNA SOCS1 dan populasi makrofag terhadap progresivitas penyakit yang berhubungan dengan Benign Prostate Hyperplasia (BPH), High Grade Prostatic Intraepithelial Neoplasia (HGPIN), dan Prostate Adenocarcinoma (PRAD). Metode: Penelitian ini merupakan penelitian potong lintang dengan 3 kelompok, yaitu BPH,HGPIN, dan PRAD. Sampel jaringan didapatkan dari Tindakan TURP. Ekspresi miR-155 dianalisis menggunakan qPCR dan dikalkulasi menggunakan metode Livak. Ekspresi mRNA SOCS-1 dianalisis menggunakan reverse transcriptase PCR. Penanda pan makrofag, anti CD-68 monoclonal antibody(MoAb) digunakan untuk mendeteksi populasi makrofag pada jaringan dengan imunohistokimia. Hasil: Ekspresi miR-155 lebih tinggi pada HGPIN dibandingkan BPH dan PRAD (p=0,14). Ekspresi mRNA SOCS1 pada HGPIN paling rendah diantara ketiga sampel (p=0,96). Terdapat korelasi negative antara miR-155 dan mRNA SOCS1 (p=0,02). Terdapat peningkatan persentase populasi makrofag yang signifikan pada HGPIN (6,03 persen) dibandingkan BPH (0.89 persen) dengan p=0,00. Kesimpulan: Pada penelitian ini, terdapat perubahan persentase makrofag dan miR-155 pada HGPIN. Variasi ekspresi miR-155 dan persentase populasi makrofag dapat disebabkan karena perubahan epigenetik. Oleh sebab itu, perlu penelitian lebih lanjut untuk memvalidasi hasil tersebut dan memahami kemungkinan menjadi biomarker pada penyakit prekanker pada prostat. Kata Kunci: Prostatic Intaepithelial Neoplasia, miR-155, Makrofag Abstract Background: Impaired microRNA(miR) regulation and chronic inflammation could transform tumors into carcinoma and cancer by metastasis through cellular and genomic changes. Precancerous lesions have a 33.3 percent chance of becoming cancerous. This study investigated the role of miR-155 related to SOCS1 mRNA and macrophage population in disease progression associated with Benign Prostate Hyperplasia (BPH), High-Grade Prostatic Intraepithelial Neoplasia (HGPIN), and Prostate Adenocarcinoma (PRAD). Methods: This was a cross-sectional study using three groups of samples, namely BPH, HGPIN, and PRAD. Tissue samples were obtained from TURP Action. The expression of miR-155 was analyzed using real-time qPCR and calculated using the Livak method. The expression of SOCS1 mRNA was analyzed using reverse transcriptase PCR. The macrophage pan-marker, anti-CD68 monoclonal antibody (MoAb), was used to detect macrophage population in tissues by immunohistochemistry. Results: The expression of miR-155 was higher in HGPIN than BPH and PRAD (p=0.14). The expression of SOCS1 mRNA in HGPIN was the lowest among the three samples (p=0.96). There was a negative correlation between miR-155 and SOCS1 mRNA (p=0.02). There was a significant increase in the percentage of the macrophage population in HGPIN (6.03 percent) compared to BPH (0.89 percent) with p=0.00. Conclusion: In this study, there were changes in the percentage of macrophage and miR-155 in HGPIN. The variation in miR-155 expression and the percentage of the macrophage may be caused by epigenetic changes. Therefore, further research is needed to validate these results and understand the possibility of being a biomarker in precancerous disease of the prostate. Keywords: Prostatic Intraepithelial Neoplasia, miR-155, Macrophage more...

BACKGROUND: Triple-negative breast cancer (TNBC) represents a heterogenous disease which differ in characteristic, treatment response and prognosis. We aim to perform in-depth analysis on the clinicopathologic feature and the prognostic value of basal-like and non-basal-like TNBC patients in an Indonesian tertiary hospital. METHODS: We retrospectively included patients diagnosed with TNBC between 2014-2017. Clinical variables were collected from medical record. Expression of epidermal growth factor receptor (EGFR), cytokeratin 5/6 (CK5/6), p53 mutant and androgen receptor (AR) were examined by using immunohistochemistry (IHC). RESULTS: We included 67 subjects, 67.1% were basal-like and the remaining 32.9% were non-basal-like, with mean age of 51 years old, 59.7% subjects had BMI more...

Purpose: Ovarian cancer is the most lethal of gynecological malignancies. Recently, the development of microRNA (miRNA) -based therapeutics that could impact broad cellular programs, leading to inhibition of cancer cell viability, is gaining attention in the therapeutic landscape. The therapy is based on the presence of aberrant expressions of miRNA in cancer cells. Decreasing of tumor suppressor miRNA expression causes upregulation of oncoprotein, which worsens the prognosis of the ovarian cancer. Methods: miR-155-5p mimics were carried by chitosan nanoparticles using new nanotechnology methods. Cellular uptake of miRNA was assessed by fluorescence microscope while MTT and qPCR assay were used to determine miRNA profile and the effect of CS-NP/miRNA on SKOV3 cells. Results: Results of profiling validated using quantitative realtime-polymerase chain reaction (PCR) found one of the most altered tumor suppressor miRNAs, miR-155-5p was downregulated 892.15-fold. According to bioinformatic analysis we identified the miRNA could recognize and regulate HIF1α expression. Transfection of mimics for miR-155-5p showed significantly increased miR-155-5p endogen SKOV3 expression level compared to the control group. We found differences after transfection mimics for miR-155-5p 31.5 and 63 nanoMolar. Increasing of miR-155-5p endogen lead to diminished SKOV3 viability (by 30%; more...

Globally, prostate cancer (PCA) is the second leading cause of male cancer-associated mortality. Micro-RNAs (miRNAs) are small non-coding RNAs considered promising biomarkers for diagnosis, prognosis, and treatment options. A miR-141 expression is frequently dysregulated and influences the development and progression of PCA. This study aimed to identify miR-141 expression level as a marker to differentiate PCA from another prostate anomaly, especially in Yogyakarta. Formalin-fixed paraffin-embedded (FFPE) tissues for each three groups: benign prostatic hyperplasia/BPH, high-grade prostatic intraepithelial neoplasia/HGPIN, and PCA (n=7/group) were stored in a commercial clinical laboratory in Yogyakarta. The total RNA was extracted from FFPE sections using miRNeasy FFPE kit, followed by the quantification of miR-141-3p expression level by RT-PCR. The result showed that miR-141 relative expression level on PCA was higher than other groups and significantly different (P more...

Objective: External counterpulsation (ECP) provides long-term benefits of improved anginal frequency and exercise tolerance in patients with refractory angina (RA). This is postulated as a result of improved angiogenesis and endothelial function through an increase in shear stress. Angiogenesis is mainly represented by vascular endothelial growth factor-A (VEGF-A) and its receptor, vascular endothelial growth factor receptor-2 (VEGFR-2). The microRNA-92a (miR-92a) is a flow-sensitive miRNA that regulates atherosclerosis and angiogenesis in response to shear stress. Thus, ECP beneficial effect might be achieved through interaction between VEGF-A, VEGFR-2, and miR-92a. This study aims to evaluate the ECP effect on VEGF-A, VEGFR-2, and miR-92a in patients with RA in a sham-controlled manner.Methods: This was a randomized sham-controlled trial, enrolling 50 patients with RA who have coronary artery disease (CAD). Participants were randomized (1:1 ratio) to 35 sessions of either ECP (n = 25) or sham (n = 25), each session lasting for 1 h. Plasma levels of VEGF-A and VEGFR-2 were assayed by the ELISA technique. The quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to measure miR-92a circulating levels in plasma.Result: External counterpulsation significantly preserved VEGF-A and VEGFR-2 level compared to sham [ΔVEGF-A: 1 (−139 to 160) vs.−136 (−237 to 67) pg/ml, p = 0.026; ΔVEGFR-2: −171(-844 to +1,166) vs. −517(−1,549 to +1,407) pg/ml, p = 0.021, respectively]. Circulating miR-92a increased significantly in ECP [5.1 (4.2–6.4) to 5.9 (4.8–6.4), p < 0.001] and sham [5.2 (4.1–9.4) to 5.6 (4.8–6.3), p = 0.008] post-intervention. The fold changes tended to be higher in ECP group, although was not statistically different from sham [fold changes ECP = 4.6 (0.3–36.5) vs. sham 2.8 (0–15), p = 0.33)].Conclusion: External counterpulsation improved angiogenesis by preserving VEGF-A and VEGFR-2 levels. Both ECP and sham increased miR-92a significantly, yet the changes were not different between the two groups. (Study registered on www.clinicaltrials.gov, no: NCT03991871, August 8, 2019, and received a grant from the National Health Research and Development of Ministry of Health of Indonesia, No: HK.02.02/I/27/2020). more...

BACKGROUND: The diagnosis of tuberculosis (TB) in children is challenging by the absence of a practical gold standard. Interferon (IFN)-ginducible protein 10 (IP-10) is a chemokine that may serve as the leading candidate marker in child TB diagnosis. The aim of this study is to assess the diagnostic value of IP-10 in the diagnosis of TB in children. METHODS: We recruited eligible symptomatic and asymptomatic children aged more...

Abstract Background Squamous cell carcinoma of the oral cavity (OSCC) is the sixth most common malignancy. Surgery is mainstay treatment for oral cancers. Surgery in locally advanced OSCC presents many challenges primarily because the head and neck have critical structures that can be damaged by tumor or treatment. It is thought that neoadjuvant chemotherapy (NC) in locally advanced OSCC is able to shrink tumor size. Chemoresistancy is a problem due to hypoxic microenvironment characterized by increased expression of HIF-1α. It is also regulated by miR-210 as well as increased expression of CD44 and CD133. Melatonin has a powerful antioxidant and oncostatic effects that are expected to improve tumor hypoxia and clinical response. Fifty patients with OSCC were included and randomized. miR-210 and CD44 expression were measured before and after intervention using qRT-PCR absolute quantification, and clinical response was evaluated according to RECIST 1.1 criteria. This study aims to determine the effect of melatonin in improving the clinical response of patients with locally advanced oral squamous cell carcinoma (OSCC) after neoadjuvant chemotherapy to miR-210 and CD44 expression. Results Melatonin administration reduced miR-210 levels but not significant (p = 0.767). CD44 expression also decreased in the melatonin group compared with placebo yet was not significant (p = 0.103). There was a decrease in the expression of miR-210 and CD44 followed by a decrease in the percentage of residual tumor but not significant (p = 0.114). Conclusion In OSCC, the addition of 20-mg melatonin to neoadjuvant chemotherapy (NC) reduced the expression of miR-210 and CD44 and decreased the percentage of tumor residue; however, no statistically significant result was observed. Trial registration This study is registered to ClinicalTrials.gov under trial registration number: NCT04137627 with date of registration on October 22, 2019—retrospectively registered, accessible from: https://clinicaltrials.gov/ct2/show/NCT04137627 more...

Red dragon fruit is one of the popular fruits that have been widely used both for consumption and food coloring. The red dragon fruit peel and flesh contain various antioxidant compounds that can be used as pharmaceutics and nutraceuticals. The objective of this study was to determine the effect of various extract preparations of the peel and the flesh of red dragon fruit on the viability of normal human fibroblasts. Seven conditions of peel and flesh extracts were prepared as follows, i.e. dried peel ethanolic extract, fresh blended peel ethanolic extract, dried flesh, fresh blended flesh ethanolic extract, blended fresh flesh, filtrate of pressed flesh, and pomace of pressed flesh. Each sample preparation was tested for its effect on the viability of normal human fibroblasts using MTT assay. Results showed that dried peel ethanolic extract reduce cell viability. Red dragon fruit flesh extracts caused no significant effect on the fibroblast viability. In conclusion, the fruit flesh extracts are relatively safer to normal cells than the peel extracts. IC50 value of the ethanolic extract of dried peel was 55.38±3.85 µg/mL, while the IC50 value of various types of flesh extract were more than 500 µg/mL. more...

Carica papaya leaves are one of the vegetables consumed by Indonesian people, especially in Java Island. Carica papaya is easy to grow in Indonesia and has many variants, so, Carica Papaya leaves is a local potent to be developed for functional food and nutraceutical. The aim of this study was to investigate antioxidant activity and total flavonoids of Carica papaya leaves with different varieties, maturity and solvent. Carica papaya leaves (CPL) was firstly extracted by methanol to select two CPLs with high antioxidant capacity and total flavonoid. The two selected CPLs were further tested with different ages mainly young and mature leaves. One selected CPL was further tested with different extraction solvents. Antioxidant activity was determined by 2.2 diphenyl-1-picrylhydrazyl, DPPH and Ferric reducing antioxidant power, FRAP. This study used five varieties of Carica papaya leaves, namely Bangkok, California, Purple, Golden and Grendel. The result showed that Golden and Grendel varieties had a higher percentage of radical scavenging property than the others, which was 78.37% and 77.40% by the DPPH method. Grendel and Purple had a higher percentage of radical scavenging property, which was 45.82 and 34.32 mmol/mg. Grendel and Purple had a higher total flavonoid property, which was 50.33 and 46.02 µg/g. Mature leaves had a higher percentage of radical scavenging property than young leaves by DPPH and FRAP methods. Mature leaves had a higher total flavonoid property than young leaves in both Grendel and Purple. Grendel had a higher antioxidant activity and a higher total flavonoid property than Purple. Grendel with water extraction had a higher antioxidant activity by DPPH and FRAP methods. The total flavonoid of Grendel papaya leaves’ extract with water extraction was lower than ethanol 70% and methanol. more...

AIM: To investigate neurofibromatosis type 2 (NF2) gene mutation at mRNA levels in sporadic orbitocranial meningioma and its association with progesterone receptor (PR) mRNA expression. METHODS: This was a case-control study. Thirty-four sporadic meningioma patients with no familial NF2-related meningioma history were recruited. They were interviewed for their obstetric, gynecologic, and contraception history. PR investigation was performed with real-time polymerase chain reaction (PCR). NF2 mutation was investigated using Qbiomarker Somatic Mutation PCR Assay at NF2 mRNA level after its cDNA extraction (four mRNA mutation cytoband coordinates for nucleotide change: c.634C>T/p.Q212, c.655G>A/p.V219M, c.784C>T/p.R262 and c.1228C>T/p. Q410). RESULTS: After mutation analysis at mRNA level, NF2 gene mutation was found in 35.29% patients. Non-mutation group was strongly associated with exogenous hormonal exposure (non-mutation vs mutation: 95.5% vs 83.3%, P more...

Objectives: Monitoring the spread of the G614 in specific locations is critical as this variant is highly transmissible and can trigger the emergence of other mutations. Therefore, a rapid and accurate method that can reliably detect the D614G mutation will be beneficial. This study aims to analyze the potential use of the two-step Reverse Transcriptase quantitative polymerase chain reaction - high resolution melting analysis (RT-qPCR-HRM) to detect a specific mutation in the SARS-CoV-2 genome. Methods: Six SARS-CoV-2 RNA samples were synthesized into cDNA and analyzed with the qPCR-HRM method in order to detect the D614G mutation in Spike protein of SARS-CoV-2. The primers are designed to target the specific Spike region containing the D614G mutation. The qPCR-HRM analysis was conducted simultaneously, and the identification of the SARS-CoV-2 variant was confirmed by conventional PCR and Sanger sequencing methods. Results: The results showed that the melting temperature (Tm) of the D614 variant was 79.39 ± 0.03 °C, which was slightly lower than the Tm of the G614 variant (79.62 ± 0.015 °C). The results of the HRM analysis, visualized by the normalized melting curve and the difference curve were able to discriminate the D614 and G614 variant samples. All samples were identified as G614 variants by qPCR-HRM assay, which was subsequently confirmed by Sanger sequencing. Conclusions: This study demonstrated a sensitive method that can identify the D614G mutation by a simple two-step RT-qPCR-HRM assay procedure analysis, which can be useful for active surveillance of the transmission of a specific mutation. more...

Objective: We determine the utility of Prostate Specific Antigen (PSA) for predicting the presence of skeletal metastasis on Bone Scan (BS) in prostate cancer patients. Material & Methods: Retrospective analysis of medical records of 70 consecutive prostate cancer patients subjected to bone scan during the last 2 years was done. 5 cases were excluded due to the following reasons: Serum PSA not available, hormonal or other therapy given prior to serum PSA measurement, and/or Bone Scan, and symptomatic for bone metastasis. In remaining 65 cases, PSA value and bone scan were evaluated. Results: BS was found to be positive in 20/65 (31%) and negative in 45(69%) patients. 24 (37%) had serum PSA > 100 ng/ml, 25 (38.5%) had PSA of 20‐100 ng/ml and only 16 (24.5%) had PSA < 20 ng/ml. Conclusion: Serum PSA < 20 ng/ml have high predictive value in ruling out skeletal metastasis. Our data are in corroboration with results from previous studies that BS should be performed only if PSA > 20 ng/ml. Using this cut‐off, unnecessary investigation can be avoided. Avoiding BS asymptomatic in this group of patients would translate into a significant cost‐saving and reduction in their psychological and physical burden. more...

Background: Noncontact Electro Capacitive Cancer Therapy (ECCT) is a novel treatment modality in cancer. Chemokine (C-C motif) ligand 2 (CCL2) has a major role in the outgrowth of metastatic breast cancer. Interleukin 18 (IL18) plays a role in macrophage alteration, which leads to excessive angiogenesis. This study aims to elaborate on the association of CCL2, IL18, IL23α, and TNF-α (tumor necrosis factor-alpha) expression with the anti-proliferative effect of ECCT in rat breast tumor tissue. Methods: Low intensity (18 Vpp) and intermediate frequency (150 kHz) alternating current-electric field (AC-EF) between two capacitive electrodes were exposed as external EF to a rat cage. Twenty-four rats were divided into four groups of six replicates. Breast tumor tissues were collected from 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rats. Two groups were non DMBA-induced rats without ECCT exposure (NINT) and with (NIT). The other two groups were DMBA-induced rats without ECCT exposure (INT) and with (IT). Mammary glands and breast tumor tissues were collected from each group and preserved. Hematoxylin-eosin and immunohistochemistry staining were performed on paraffin sections of tissues using anti-PCNA, anti-ErbB2, anti-Caspase3, and anti-CD68. CCL2, IL18, IL23α, and TNF-α mRNA relative expressions were analyzed using qRT-PCR. Results: ECCT exposure may cause the reduction of PCNA protein expression as well as ErbB2 on breast tumor tissues, but it causes the increase of Caspase3 and macrophage CD68 protein. In rat breast tumor tissues of IT groups, the mRNA expression of CCL2 and IL18 are significantly down-regulated, in contrast with the up-regulated expression of these cytokines in tumor tissues of the INT group. IL23α and TNF- α expression remained similar in both groups. Conclusion: CCL2 and IL18 expressions have an association with the inhibition of breast tumor cell proliferation affected by ECCT exposure more...

Background: Noncontact Electro Capacitive Cancer Therapy (ECCT) is a novel treatment modality in cancer. Chemokine (C-C motif) ligand 2 (CCL2) has a major role in the outgrowth of metastatic breast cancer. Interleukin 18 (IL18) plays a role in macrophage alteration, which leads to excessive angiogenesis. This study aims to elaborate on the association of CCL2, IL18, IL23α, and TNF-α (tumor necrosis factor-alpha) expression with the anti-proliferative effect of ECCT in rat breast tumor tissue. Methods: Low intensity (18 Vpp) and intermediate frequency (150 kHz) alternating current-electric field (AC-EF) between two capacitive electrodes were exposed as external EF to a rat cage. Twenty-four rats were divided into four groups of six replicates. Breast tumor tissues were collected from 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rats. Two groups were non DMBA-induced rats without ECCT exposure (NINT) and with (NIT). The other two groups were DMBA-induced rats without ECCT exposure (INT) and with (IT). Mammary glands and breast tumor tissues were collected from each group and preserved. Hematoxylin-eosin and immunohistochemistry staining were performed on paraffin sections of tissues using anti-PCNA, anti-ErbB2, anti-Caspase3, and anti-CD68. CCL2, IL18, IL23α, and TNF-α mRNA relative expressions were analyzed using qRT-PCR. Results: ECCT exposure may cause the reduction of PCNA protein expression as well as ErbB2 on breast tumor tissues, but it causes the increase of Caspase3 and macrophage CD68 protein. In rat breast tumor tissues of IT groups, the mRNA expression of CCL2 and IL18 are significantly down-regulated, in contrast with the up-regulated expression of these cytokines in tumor tissues of the INT group. IL23α and TNF- α expression remained similar in both groups. Conclusion: CCL2 and IL18 expressions have an association with the inhibition of breast tumor cell proliferation affected by ECCT exposure more...

The outcome of inoperable rectal cancer treatment by chemotherapy, radiotherapy, and targeted therapy are still unfavorable. Carbogen is a combination of 98% oxygen and 2% carbon dioxide proven effective as chemoradiosensitizer. The aim of this study is to know the effect of concurrent carbogen and chemoradiotherapy in locally advanced rectal cancer by measuring the shrinkage of the tumor volume. The design of this study was randomized true experimental 2 groups pre and post-test-controlled design. Samples were patients with locally advanced rectal cancer. MRI of the pelvis before and 4-8 weeks after the chemoradiation were examined. A total of 28 subjects were randomized to 14 patients who received concurrent chemoradiation with carbogen (treatment group) and 14 patients chemoradiation (control) The tumor shrinkage in the treatment group (13.08 to 6.08 cm3) was significantly higher compared to the control group (18.00 to 12.83 cm3). Supplementation of carbogen to standard treatment chemoradiation for locally advanced rectal cancer significantly shrinkage the tumor volume. more...

Abstract Objective The identification of new genetic-associated risk factor of ischemic stroke could improves strategies for stroke prevention. This study aims to identify insulin receptor substrate 1 (IRS-1) gene polymorphism Gly972Arg as the risk factor for ischemic stroke among Indonesian subjects. The case–control study was conducted by matching the gender and race on 85 cases of patients with ischemic stroke and 86 healthy non-stroke control subjects. Ischemic stroke was established by the complete neurology examination and brain computed tomography scan or magnetic resonance imaging. Polymerase chain reaction–Restriction Fragment Length Polymorphism was performed to analyze IRS-1 gene Gly972Arg genotype. Results There were 85 ischemic stroke cases and 86 control subjects. The distribution of nucleotide IRS-1 gene polymorphism Gly972Arg in the ischemic stroke vs health controls for GG were 32.2% vs 41.5%, for GR were 16% vs 7.6%, and for RR were 0.5% vs 1.9%. IRS-1 gene polymorphism Gly972Arg was found as significant risk factor for ischemic stroke [odds ratio of 2.6 (1.27–5.27); CI 95%, p = 0.008]. Conclusively, the IRS-1 gene polymorphism Gly972Arg should be considered as an important factor in the prevention and treatment of ischemic stroke. more...

Ovarian cancer is a lethal disease. One of the problems faced by patients with ovarian cancer is the lack of symptoms in its early stages, which results in it only being detected when it is at an advanced stage. Therefore, there is an urgent need for biomarkers that can predict ovarian cancer precisely. The purpose of this study was to determine the expression of microRNA-21 as a predictive biomarker candidate in both early- and advanced-stage ovarian cancer. This was a cross-sectional study using the blood plasma of 21 healthy control subjects and 37 blood plasma samples from patients with ovarian cancer. Blood plasmas were collected, from which the RNA was isolated. Based on the RNA, the cDNA was synthesized and run through qPCR, the results of which were analyzed using the Livak method. The results showed an upregulation of microRNA-21 in the advanced stage by 2.14 fold compared with the early stage, and 6.13 fold compared with the healthy controls (p < 0.05). The upregulation of microRNA-21 in early-stage ovarian cancer was 2.86 fold compared with the healthy control subjects (p < 0.05). In addition, there was an increase in the expression of microRNA-21 in ovarian cancer by 4.14 fold compared with the healthy controls (p < 0.05). Based on these results, it can be concluded that the expression of microRNA 21 upregulated with the severity of the disease. more...

Background: Noncontact Electro Capacitive Cancer Therapy (ECCT) is a novel treatment modality in cancer. Chemokine (C-C motif) ligand 2 (CCL2) has a major role in the outgrowth of metastatic breast cancer. Interleukin 18 (IL18) plays a role in macrophage alteration, which leads to excessive angiogenesis. This study aims to elaborate on the association of CCL2, IL18, IL23α, and TNF-α (tumor necrosis factor-alpha) expression with the anti-proliferative effect of ECCT in rat breast tumor tissue. Methods: Low intensity (18 Vpp) and intermediate frequency (150 kHz) alternating current-electric field (AC-EF) between two capacitive electrodes were exposed as external EF to a rat cage. Twenty-four rats were divided into four groups of six replicates. Breast tumor tissues were collected from 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rats. Two groups were none DMBA-induced rats without ECCT exposure (NINT) and with (NIT). The other two groups were DMBA-induced rats without ECCT exposure (INT) and with (IT). Mammary glands and breast tumor tissues were collected from each group and preserved. Hematoxylin-eosin and immunohistochemistry staining were performed on paraffin sections of tissues using anti-PCNA, anti-ErbB2, anti-Caspase3, and anti-CD68. CCL2, IL18, IL23α, and TNF-α mRNA relative expressions were analyzed using qRT-PCR. Results: ECCT exposure may cause the reduction of PCNA protein expression as well as ErbB2 on breast tumor tissues, but it causes the increase of Caspase3 and macrophage CD68 protein. In rat breast tumor tissues of IT groups, the mRNA expression of CCL2 and IL18 are significantly down-regulated, in contrast with the up-regulated expression of these cytokines in tumor tissues of the INT group. IL23α and TNF- α expression remained similar in both groups. Conclusion: CCL2 and IL18 expressions have an association with the inhibition of breast tumor cell proliferation affected by ECCT exposure more...

ABSTRACT: Background: The increasing rate of cancer chemoresistance and adverse side effects of therapy have led to the wide use of various chemotherapeutic combinations in cancer management, including lymphoid malignancy. Objective: We investigated the effects of a combination of 1,3,6-trihydroxy-4,5,7-trichloroxanthone (TTX) and doxorubicin on the Raji lymphoma cell line. Methods: Raji cells were treated with different concentrations of TTX, doxorubicin, or combinations thereof. Cancer cell growth inhibition was evaluated using 3-(4,5-dimethyltiazol-2-yl)-2,5- diphenyltetrazolium bromide/MTT assay to determine the half-maximal inhibitory concentration. Combination index values were calculated using CompuSyn (ComboSyn, Inc, Paramus, NJ). Molecular docking was conducted using a Protein-Ligand ANT System. Results: The mean (SD) half-maximal inhibitory concentration values of TTX and doxorubicin were 15.948 (3.101) µM and 25.432 (1.417) µM, respectively. The combination index values of the different combinations ranged from 0.057 to 0.285, indicating strong to very strong synergistic effects. The docking study results reveal that TTX docks at the active site of Raf-1 and c-Jun N-kinase receptors with predicted free energies of binding of −79.37 and −75.42 kcal/mol, respectively. Conclusions: The xanthone-doxorubicin combination showed promising in vitro activity against lymphoma cells. The results also indicate that the TTX and doxorubicin combination's effect was due to the interaction between TTX with Raf-1 and c-Jun N-kinase receptors, 2 determinants of doxorubicin resistance progression. more...

Background: Osteoblast and osteoclast are the important factor in periodontal tissue remodeling for the orthodontic treatment success. Resorption process takes place in compression area by osteoclast and apposition in the tension area by osteoblast. In general hormone condition and age affect remodeling process. Estrogen has a high contribution in remodelling process and decreased in elderly individual such as menopausal women. Soybean contains isoflavone genistein which has similar structure and activity to estrogen. Many researchers indicate that isoflavone genistein not only has an inhibitor effect in osteoporosis but also has estrogenic and antiestrogenic effect as well. Purpose: The study aimed to investigate the effect of soybean isoflavone genistein administration on osteoblast and osteoclast cells number in orthodontic tooth movement of young and old guinea pigs. Method: The research was quasi-experimental study with post test only with control design. The experimental animals were 24 male guinea pigs that divided into: young guinea pigs (±4 months old) and old guinea pigs (±2.5 years old). Each group was divided into 4 subgroups for receiving the treatment namely; control, orthodontic treatment, genistein treatment and orthodontic+genistein treatment. All of the subjects were sacrificed at day 7 and the specimens were histologically analyzed using tartrate resistance acid phosphatase (TRAP) and hematoxylin eosin (HE) staining and observed using microscope that connected to obtilab and an image raster program. Result: U Mann-Whitney statistical analysis showed there were significant differences in osteoblast cell numbers; between orthodontic treatment and orthodontic+genistein treatment in the old guinea pigs (p=0.004); between orthodontic treatment in the young guinea pig and orthodontic+genistein treatment in the old guinea pig (p=0.016); between orthodontics treatment and orthodontic+genistein treatment in the young guinea pigs (p=0.025). U Mann-Whitney statistical analysis showed there were significant differences in osteoclast cell numbers: between the orthodontic treatment in the old guinea pig and orthodontics+genistein treatment in the young guinea pigs (p=0.007); between orthodontic treatment group in the young guinea pigs and orthodontics+genistein treatment in the old guinea pigs; between orthodontic treatment and orthodontic+ genistein treatment in the young guinea pigs (p=0.007). All groups administered by genistein the numbers of osteoblast in the surrounding of the tension sites increased, while in the surrounding of the compression sites had less osteoclasts; even, there were no osteoclasts found in some samples. Conclusion: Soybean isoflavone genistein administration on orthodontic tooth movement increased osteoblast numbers in the tension sides and decreased osteoclast numbers in the compression sides. more...

Background: The pathogenesis of meningioma in females and its association with exogenous progesterone is remained unclear. This study was aimed to examine expression of Progesterone receptor (PR) and Neurofibromatosis-2 (NF2) and assess their relationships to history of exogenous progesterone use and risk of meningioma.Methods: Our study was a case-control study that involves 115 females, 40 cases who diagnosed with orbito-cranial meningioma and 75 controls of healthy, that has been presented in previous study. The demographic characteristics, reproductive factors, and history of progesterone use were obtained in–depth face-to-face interviews. PR and NF2 mRNA were assessed by real-time quantitative polymerase chain reaction (RT-qPCR) on serum specimens.Results: The mean age of participants in cases vs. controls were 46.6 ± 6.2 vs. 46.5 ± 7.45 (P = 0.969). The expression of PR and NF2 in cases was significantly lower than in controls. The longer duration of progesterone exposure was significantly associated with lower expression of PR and NF2. Significant association between lower expression of PR (OR 11.7; 95% CI 4.17–32.9; P < 0.001 comparing the lowest quartile vs. 3 highest quartile of PR) and NF2 (OR 4.23; 95% CI 1.85–9.67; P = 0.001 comparing the 2 lowest quartiles vs. 2 highest quartiles) with increased risk of meningioma were also reported.Conclusion: In this study we showed that the longer the exposure to exogenous progesterone, the lower the expression of PR and NF2 mRNA in the serum. Low expression of PR and NF2 were associated with higher risk of meningioma, suggesting that low PR expression and inactivation of NF2 might play a key role in progesterone-associated meningioma tumorigenesis and may be potential clinical marker for females at higher risk of meningioma. more...