Expression and clinical significance of PRC1 in soft tissue sarcoma based on data mining and effect of PRC1 on the growth of liposarcoma / 西安交通大学学报(医学版)

Objective: To investigate the expression and clinical significance of protein regulator of cytokinesis 1 (PRC1) in soft tissue sarcoma (STS) and the effects of down-regulating the expression of PRC1 on the proliferation and cell cycle of human liposarcoma cell by data mining. Methods: Oncomine database was used to analyze the expression of PRC1 in STS tissue and normal tissue, which was then verified by TCGA database. The prognosis data downloaded from OncoLnc database were used to analyze the correlation between PRC1 expression and prognosis of STS patients. Meanwhile, to collect PRC1 expression in STS cells, we used publicly available data from the Cancer Cell Line Encyclopedia (CCLE) projects. String database was used to determine the co-expression molecules with PRC1 and map the gene co-expression network. The expressions of PRC1 in liposarcoma cell line SW872 and human subcutaneous preadipocytes (HPA-s) were detected by Western blotting and Real-time PCR. PRC1 was silenced in SW872 cell (SW872-siPRC1) by PRC1 target siRNA with SW872-NC cell as its control. MTT assay was used to detect the proliferation of SW872-siPRC1 and SW872-NC cells; flow cytometry was used to detect the cell cycle. Results: In the Oncomine database, 11 studies involved PRC1 expression in STS tissues and normal tissues. Compared with that of the control, the expression of PRC1 in STS tissues was significantly higher (P<0.05). The results were consistent with those in the TCGA database. The analysis using Oncomine database showed that the high expression level of PRC1 was associated with shorter overall survival (P<0.05). The analysis using CCLE database showed high expression of PRC1 in STS cells (P<0.05). The co-expression network of PRC1 was established by String database, including 11 nodes and 55 connections. PRC1 was over-expressed in liposarcoma cell line SW872. Cell proliferation curve showed that compared with that of SW872-NC cells in the control group, the proliferation of SW872-siPRC1 cells decreased significantly after 48 h and 72 h culture (P<0.05). Compared with SW872-NC cell in the control group, the G1 cell proportion of SW872-siPRC1 cells was (40.27±7.42)%, significantly lower than that of SW872-NC cells (62.01±4.89)%. The G2/M cell proportion of SW872-siPRC1 cells was (25.65±1.54)%, which was significantly higher than that of SW872-NC cells (8.17±0.96)% (both P<0.05). Conclusion: Tumor gene database mining shows that PRC1 is highly expressed in STS tissues and STS cells, which is associated with the patient's prognosis. Silencing PRC1 gene can inhibit the proliferation of liposarcoma SW872 cells and keep the cells staying in G2/M phase. PRC1 plays a role in promoting liposarcoma, which may provide a potential target for the clinical treatment and prognosis of soft tissue sarcoma, especially liposarcoma.

Transfusion Requirement in Anemia observed in Patients with Carcinoma Breast undergoing Chemotherapy

Introduction: Carcinoma Breast is the second most commonmalignancyaffecting women. Adjuvant chemotherapy is themainstay treatment modality along with surgery. Anemiaand thrombocytopenia are the hematological complicationsin patients with breast cancer who undergo chemotherapy.This study was carried out with an objective to identify theproportion of anemia and the requirement of Packed Red Cellstransfusions in these patients.Material and methods: This is ahospital based prospectivestudy, done for a period of one and a half years from January2016 to June 2017.As per the inclusion criteria125 consecutivecases who underwent chemotherapy for Carcinoma Breastat Department of Radiotherapy, Govt. Medical College,Thiruvananthapuram were included for the study. Therequirement of Packed Red Cell (PRC) was assessed in thesepatients. Data was analysed with SPSS software (version 21).Results: Among the 125 patients, 60% of patients wereanemic in prechemotherapyphase with a preponderance ofmild anemia,haemoglobin (Hb) 11-11.9gm/dl (as per theWHO classification of anemia in non pregnant females).Inpost chemotherapy phase the prevalence of anemia was94.4% withincreasing severity; majority (56%) of patients hadmoderate anemia (Hb 8-10.9gm/dl). During chemotherapy,22.4% of the study population required PRC transfusion.Conclusion: Due to the high prevalence of chemotherapyinduced anemia and itseffects on quality of life (QOL), evenmild degrees of anemia should be detected and evaluatedbefore commencing chemotherapy. PRC transfusion shouldbe reserved for patients with severe anemia

In-house standardization and validation of a multiplex RT-PCR assay for the detection of 13 respiratory viruses / Estandarización interna y validación de un ensayo RT-PRC múltiple para la detección de 13 virus respiratorios

Background. Multiplex real time PCR is increasingly used to diagnose respiratory viruses and has shown to be superior to traditional methods, such as culture and antigen detection. Objective. Standardization and validation of a multiplex real-time PCR assay for the detection of 13 respiratory viruses. Methods. The assay was validated using RNA control targets and comparing results to single-target PCR's. Results. Using RNA controls the multiplex format was found to be as sensitive and specific as the single-target PCRs, and no competition was observed between targets. The efficiencies for most of the reactions were approximately 90%, but a lower efficiency was found for Parainfluenza 2 with a rate of amplification in each cycle of 86.63%. On the other hand, a higher efficiency was observed in respiratory syncytial virus A and respiratory syncytial virus B ((93.07% each). Conclusion: This multiplex RT-PCR format shows an adequate efficiency, demonstrating an excellent sensitivity, specificity and repeatability for all the studied respiratory viruses.

Antecedentes. PRC múltiple en tiempo real es usada cada vez más para el diagnóstico de virus respiratorios y ha mostrado ser superior a metodos tradicionales, como cultivo y detección de antígeno. Objetivo. Estandarizar y validar una PRC múltiple en tiempo real para la detección de 13 virus respiratorios. Metodos. El ensayo fue realizado usando blanco de RNA control y comparando los resultados a blancos únicos de PCR. Resultados. Usando el RNA control, el formato de multiplex era tan sensible y específico como la PCR. Las eficiencias para la mayoría de las reacciones de aproximadamente el 90%, pero una eficiencia baja fue encontrada para influenza 2 con una tasa de amplificación en cada ciclo de 86.63%. Por otra parte, una mayor eficiencia fue observada en virus sincitial respitario A y B (93, 67% cada uno). Conclusión. Este formato RT-PCR múltiple muestra una adecuada eficiencia, demostrando un excelente especificidad y reproducibilidad para todos los virus respiratorios estudiados.

Clinical Application of the Korean Personality Rating Scale for Children in Attention-Deficit Hyperactivity Disorder

OBJECTIVES: The objective of this study was to compare the Korean Personality Rating Scale for Children (K-PRC) profile between children with attention-deficit hyperactivity disorder (ADHD) and typically developing children. We also aimed to investigate the association of K-PRC and ADHD symptoms. METHODS: Ninety-nine youth (age 8.3+/-2.4 years, 72 boys) with ADHD and 84 controls (age 9.2+/-2.5 years, 43 boys) were recruited from the Department of Pediatric Psychiatry of the Asan Medical Center Children's Hospital. Diagnoses of ADHD and comorbid psychiatric disorders were confirmed with the Kiddie-Schedule for Affective Disorders and Schizophrenia-Present and Lifetime Version (K-SADS-PL). The parents of the subjects completed the ADHD rating scale, and K-PRC. Independent t-tests, analysis of covariance, partial correlation analyses, and Mc Nemar test were used for analysis. RESULTS: Children and adolescents with ADHD showed higher K-PRC scores in verbal development, physical development, depression, delinquency, hyperactivity, family dysfunction and psychoticism. Delinquency and hyperactivity were significantly correlated with parent-rated ADHD rating scales and ADHD scores on K-SADS-PL. The hyperactive/impulsive and combined subtypes showed higher scores on hyperactivity and delinquency than the inattentive subtype, and the inattentive subtype showed higher scores on depression and social dysfunction of the K-PRC. CONCLUSION: Our results suggest that K-PRC could be used to comprehensively evaluate symptoms, combined psychopathologies, developmental delay and family dysfunction of children with ADHD.

Regulation and Role of EZH2 in Cancer / Journal of the Korean Cancer Association, 대한암학회지

Polycomb repressive complex 2 (PRC2) is the epigenetic regulator that induces histone H3 lysine 27 methylation (H3K27me3) and silences specific gene transcription. Enhancer of zeste homolog 2 (EZH2) is an enzymatic subunit of PRC2, and evidence shows that EZH2 plays an essential role in cancer initiation, development, progression, metastasis, and drug resistance. EZH2 expression is indeed regulated by various oncogenic transcription factors, tumor suppressor miRNAs, and cancer-associated non-coding RNA. EZH2 activity is also controlled by post-translational modifications, which are deregulated in cancer. The canonical role of EZH2 is gene silencing through H3K27me3, but accumulating evidence shows that EZH2 methlyates substrates other than histone and has methylase-independent functions. These non-canonical functions of EZH2 are shown to play a role in cancer progression. In this review, we summarize current information on the regulation and roles of EZH2 in cancer. We also discuss various therapeutic approaches to targeting EZH2.

Avalição da qualidade microbiológica de ovos proveniente de criação tipo “caipira” e de granja de produção comercial / Evaluation of the microbiological quality of eggs from breeding type \"caipira\" farm and commercial production

O presente trabalho teve como objetivo avaliar o grau de contaminação microbiana de ovos provenientes de criação caipira e de granja de produção comercial. Foram utilizados 120 ovos, de cinco dias diferentes de postura. A avaliação microbiológica com as contagens de bactérias mesófilas aeróbias, bolores e leveduras, foram conduzidas com a análise da casca e do conteúdo interno dos ovos.Observou-se que o grau de contaminação no ovo proveniente da criação do tipo caipira foi maior do que o proveniente de produção comercial, mas a presença de Salmonella spp. foi negativa para todas as amostras. Os valores do pH de ovos “caipiras” e de granja foram em média respectivamente de 9,1 e 9,2 no albúmen e de 6,7 e 6,9 na gema.

Recrutamento do complexo repressivo polycomb 2 pelo RNA não codificador longo antissenso ANRASSF1 modula a expressão do gene RASSF1A e a proliferação celular / Recruitment of polycomb repressive complex 2 by intronic long noncoding RNA ANRASSF1 modulates RASSF1A expression and cell proliferation

O gene supressor tumoral RASSF1A tem sido associado com redução da proliferação celular em diversos tumores. Sua expressão é regulada por eventos epigenéticos que envolvem o complexo repressivo polycomb (PRC2), no entanto os mecanismos moleculares da modulação do recrutamento deste modificador epigenético para este locus ainda são desconhecidos. Neste trabalho identificamos e caracterizamos ANRASSF1, um RNA não codificador longo (lncRNA) intrônico unspliced, que é transcrito na fita oposta do gene RASSF1A, em várias linhagem celulares e tecidos, e se liga a PRC2. ANRASSF1 é transcrito pela RNAPII, possui cap-5´ e cauda poli-A, além de localizar-se no núcleo e possuir uma meia-vida em média quatro vezes menor comparada com outros lncRNAs ligados à PRC2. A super-expressão ectópica de ANRASSF1 reduziu os níveis de RASSF1A e aumentou a taxa de proliferação em células HeLa, enquanto seu silenciamento provocou efeito oposto. Essas mudanças nos níveis de ANRASSF1 não afetaram a abundância da isoforma RASSF1C em nenhuma das condições. A super-expressão de ANRASSF1 provocou um grande aumento tanto da ocupação de PRC2 como da marca de histona repressiva H3K27me3 especificamente na região promotora RASSF1A. Nenhum efeito da super-expressão de ANRASSF1 foi detectado na ocupação de PRC2 e na histona H3K27me3 nas regiões promotoras de RASSF1C e de outros quatro genes vizinhos, incluindo dois genes supressores tumorais bem caracterizados. Além disso, foi demonstrado que ANRASSF1 forma um híbrido de RNA/DNA e recruta SUZ12, um componente do PRC2, para o promotor de RASSF1A. Notavelmente, foi detectado pelo ensaio de RNase-ChIP que a degradação de ANRASSF1 diminui a ocupação de PRC2 neste promotor. Esses resultados demonstram um novo mecanismo de repressão epigenética do supressor tumoral RASSF1A, envolvendo um lncRNA unspliced antissenso, onde ANRASSF1 reprime seletivamente a expressão da isoforma de RASSF1 que sobrepõe o transcrito antissenso de modo local e específico...

Tumor-suppressor RASSF1A gene down-regulation has been implicated in increasing cell proliferation in several tumors. Its expression is regulated by epigenetic events involving polycomb repressive complex 2 (PRC2), however the molecular mechanisms modulating recruitment of this epigenetic modifier to the locus remain largely unknown. Here, we identify and characterize ANRASSF1, an endogenous unspliced long noncoding RNA (lncRNA) that is transcribed from the opposite strand of RASSF1 gene in several cell lines and tissues, and binds to PRC2. ANRASSF1 is transcribed by RNA Polymerase II, 5'-capped, polyadenylated, displays nuclear localization, and has on average a four-fold shorter half-life compared to other lncRNAs that bind PRC2. ANRASSF1 ectopic overexpression decreases RASSF1A abundance and increases the proliferation rate of HeLa cells, whereas its silencing causes opposite effects. These changes in NRASSF1 levels do not affect RASSF1C isoform abundance. ANRASSF1 overexpression causes a marked increase both in PRC2 occupancy and in histone H3K27me3 repressive mark specifically at the RASSF1A promoter region. No effect of ANRASSF1 overexpression is detected on PRC2 occupancy and on histone H3K27me3 at the promoter regions of RASSF1C and of four other neighbor genes, including two well-characterized tumor suppressor genes. Additionally, we demonstrate that ANRASSF1 forms an RNA/DNA hybrid, and recruits SUZ12, a PRC2 component, to the RASSF1A promoter. Notably, depletion of ANRASSF1 disrupts SUZ12 occupancy on RASSF1A promoter as measured by RNAse-ChIP assay. Together, these results show a new mechanism of epigenetic repression of RASSF1A tumor suppressor gene involving an antisense unspliced lncRNA, in which ANRASSF1 selectively represses expression of the RASSF1 isoform overlapping the antisense transcript in a location-specific manner. In a broader perspective, our findings suggest that other non-characterized unspliced intronic lncRNAs transcribed in the...

Transcriptional targeting of gene expression in breast cancer by the promoters of protein regulator of cytokinesis 1 and ribonuclease reductase 2

For cancer gene therapy, cancer-specific over-expression of a therapeutic gene is required to reduce side effects derived from expression of the gene in normal cells. To develop such an expression vector, we searched for genes over-expressed and/or specifically expressed in cancer cells using bioinformatics and have selected genes coding for protein regulator of cytokinesis 1 (PRC1) and ribonuclease reductase 2 (RRM2) as candidates. Their cancer-specific expressions were confirmed in both breast cancer cell lines and patient tissues. We compared each promoter's cancer-specific activity in the breast normal and cancer cell lines using the luciferase gene as a reporter and confirmed cancer-specific expression of both PRC1 and RRM2 promoters. To test activities of these promoters in viral vectors, the promoters were also cloned into an adeno-associated viral (AAV) vector containing green fluorescence protein (GFP) as the reporter. The GFP expression levels by these promoters were various depending on cell lines tested and, in MDA-MB-231 cells, GFP activities derived from the PRC1 and RRM2 promoters were as strong as that from the cytomegalovirus (CMV) promoter. Our result showed that a vector containing the PRC1 or RRM2 promoter could be used for breast cancer specific overexpression in gene therapy.

Sequencing of Flic genes of Salmonella typhi, S. paratyphi A, S. paratyphi B, S. paratyphi C, and S. typhimurium and application for PCR to differentiate them

All Flic genes of Salmonella typhi, S. paratyphi A, S. paratyphi B, S. paratyphi C, and S. typhimurium code for phase 1 of H antigen (H:1) (d, a, b, c and i antigen respectively). The genes were sequenced on Sanger's principle by automatic sequenser (ABI, 3100 Avant, Genetic Analyser). The primer pairs for the Salmonella species were designed on the basis of the collected sequences. The results showed that PCR with these primers can clearly differentiate the five Salmonella strains, especially between S. paratyphi B and S. typhimurium.

Revising Tropesis of Partial Chinese Traditional Medicine in Edition2005Pharmacopoeia of PRC(Draft Showed in Public) / 中国药房

OBJECTIVE:To get to know the revising tropesis of partial Chinese traditional medicine in Edition2005Pharmacopoeia of PPC so as to provide suggestions for the revising work.METHODS:Revising tropesis about partial new kinds and new items in Edition2005Pharmacopoeia of PRC(Draft showed in public)were analyzed in detail.RESULTS:Edition2005Pharmacopoeia of PRC adopted some new and mature skills besides the traditional method of identifying just from experience,i.e.from the appearances and quality of medicine.CONCLUSIONS:The standard of quality criteria in new edition pharma?copoeia is tend to be more rationalizing and standardizing,thus more close to the international quality criteria of natural medicinal herbs.