Study on F9 gene expression downregulation and its clinical value in hepatocellular carcinoma / 中华肝脏病杂志

Objective: To analyze the expression levels of the F9 gene and F9 protein in hepatocellular carcinoma by combining multiple gene chip data, real-time fluorescence quantitative PCR (RT qPCR), and immunohistochemistry. Additionally, explore their correlation with the occurrence and development of hepatocellular carcinoma, as well as with various clinical indicators and prognosis. Methods: The mRNA microarray dataset from the GEO database was analyzed to identify the F9 gene with significant expression differences associated with hepatocellular carcinoma. Liver cancer and adjacent tissues were collected from 18 cases of hepatocellular carcinoma. RT-qPCR method was used to detect the F9 gene expression level. Immunohistochemistry was used to detect the F9 protein level. Combined with the TCGA database information, the correlation between F9 gene expression level and prognostic and clinicopathological parameters was analyzed. The biological function of F9 co-expressed genes associated with hepatocellular carcinoma was analyzed by the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Statistical analysis was performed using Graphpad Prism software. Results: Meta-analysis results showed that the expression of the F9 gene was lower in HCC tissues than in non-cancerous tissues. Immunohistochemistry results were basically consistent with those of RT-qPCR. The data obtained from TCGA showed that the F9 gene had lower expression values in stages III-IV, T3-T4, and patients with vascular invasion. A total of 127 genes were selected for bioinformatics analysis as co-expressed genes of F9, which were highly enriched in redox processes and metabolic pathways. Conclusion: This study validates that the F9 gene and F9 protein are lower in HCC. The down-regulation of the F9 gene predicts adverse outcomes, which may provide a new therapeutic target for HCC.

Monoclonal anti-CD47 interference inpre-transfusion testing: clinical trials at home and abroad / 中国输血杂志

【Objective】 To discuss the case reports concerning anti-CD47 monoclonal antibody interfere in pre-transfusion testing, so as to find mitigation strategies for this drug interference. 【Methods】 Blood transfusion cases in clinical trials concerning CD47 mAb drugs at home and abroad were retrieved from PubMed, Medline, Web of Science, Wanfang data knowledge service platform and CNKI database. The characteristics and solutions of this drug interfering with pre-transfusion testing were analyzed. 【Results】 A total of 26 cases concerning anti-CD47 mAb interference in pretransfusion testing were retrieved, and 16 valid cases were included in this study (All received HU5F9-G4 as anti-CD47 mAb). After treatment with Hu5F9-G4, the discrepancy between forward and reverse blood typing reached 77% in pre-transfusion testing. Panagglutination was presented in antibody screenings, and all(100%) platelet antibody screenings was interfered. These results indicated that Hu5F9-G4 seriously affected the compatibility test of blood transfusion. Methods of eliminating anti-CD47 interference, as well as their advantages and disadvantages were further analyzed. 【Conclusion】 The advantages and disadvantages of eliminating anti-CD47 interference with pre-transfusion testing was analyzed according to its characteristics, which could provide reference for the laboratory testing.

Retinoic acid liposome-hydrogel: preparation, penetration through mouse skin and induction of F9 mouse teratocarcinoma stem cells differentiation

Retinoic acid (RA), a metabolite of retinol, is one of the most biologically active forms of retinoid and plays vital roles in embryonic development and in the regulation of cell proliferation and differentiation. Knowing that liposomes simulate cell membranes and that hydrogel is an ideal delivery vehicle for topical medicine, liposome-hydrogel is a novel preparation that has synergistic advantages over each component separately. Our objective was to investigate the characteristics of RA liposome-hydrogel. For quality control of the RA-loaded liposomes, we measured their morphology, particle size, Zeta-potential, and entrapment efficiency. Then we determined the viscosity of RA liposome-hydrogel. Next, the diffusion through mouse skin was explored, followed by investigation of the mRNA expression levels of Ker18, REX1, and α-FP using Q-PCR. The results showed that RA liposome-hydrogel penetrates the mouse skin effectively. The permeation rates were: Qn (%) of RA liposome-hydrogel < Qn(%) of RA-loaded liposome < Qn (%) of RA. The mRNA expression levels were dose-dependent and the effective dose decreased between vehicles due to their different release rates. F9 mouse teratocarcinoma stem cells were an ideal model to explore the mechanism of RA liposome-hydrogel in stem cell differentiation.

O ácido retinóico (RA) é um metabolito de retinol. Ele também é uma das formas mais biologicamente ativas de retinóide. Desempenha papel vital no desenvolvimento embrionário e na regulação da proliferação e diferenciação celular. Sabendo-se que lipossomas simulam a membrana das células e que hidrogel é um sistema ideal para o medicamento tópico, o lipossoma-hidrogel é uma nova preparação, que apresenta vantagens sinérgicas em relação a cada um dos componentes separados. Nosso objetivo foi investigar as características de RA lipossoma-hidrogel. A fim de controlar a qualidade do lipossoma carregado com RA, medimos morfologia, tamanho das partículas, potencial zeta e eficiência de retenção. Em seguida, determinou-se a viscosidade de RA lipossoma-hidrogel. Em seguida, avaliou-se a sua difusão através da pele de camundongos, seguida da investigação dos níveis da expressão de mRNA de Ker18, REX e de α-FP, utilizando-se Q-PCR. Os resultados mostraram que RA lipossoma-hidrogel pode penetrar na pele do camundongo de forma eficaz. As taxas de permeação foram: Qn (%) de RA lipossoma-hidrogel<Qn(%) de lipossoma RA- carregado <Qn (%) de RA. Os níveis de expressão de mRNA foram dependentes de dose e a dose efetiva diminuiu entre os veículos devido às diferentes taxas de liberação, As células estaminais de teratocarcinoma F9 de camundongo mostraram-se como modelo ideal para explorar o mecanismo de diferenciaçãode células tronco pelo RA lipossoma-hidrogel.

Application of new ERCC1 antibody for molecular diagnosis of platinum chemotherapy in non-small cell lung cancer / 中国癌症杂志

Background and purpose:High expression of excision repair cross-complementing 1 (ERCC1) is related to resistance in patients treated with platinum-containing regimens. The ERCC1 antibody 8F1 was usually used in past studies, but it was found to have no-speciifcity recently. This study aimed to investigate the predictive role of a new ERCC1 antibody 4F9 to platinum chemotherapy in non-small cell lung cancer (NSCLC) patients. Methods:Expression of ERCC1 was detected using antibody 4F9 by immunohistochemistry (IHC) in 72 NSCLC tissues. The relationship between the expression of ERCCl and the clinical pathological parameters, the efficacy of platinum chemotherapy and overall survival of patients were explored by statistical analysis. Results: The high expression of ERCCl protein was 55.5%in 72 cases. There was no signiifcant correlation between the ERCC1 expression with gender, age, pathological type, clinical stage and lymphatic metastasis (P>0.05). Patients with low expression of ERCC1 had signiifcantly higher response rates to platinum chemotherapy, longer median survival time and 2-years survival rate comparing with those with high expression of ERCC1 (62.5%vs 37.5%;22.9 vs 18.4 month;46.9%vs 37.5%), respectively (P<0.05). Conclusion:The expression analysis of ERCC1 using new ERCC1 antibody 4F9 by IHC method is helpful to assign chemotherapeutic regimen, and guide individual platinum chemotherapy for post-operation patients.

Sequence variation data of F8 and F9 genes in functionally validated control individuals: implications on the molecular diagnosis of hemophilia

BACKGROUND: The F8 and F9 genes encode for coagulation factor VIII (FVIII) and FIX, respectively, and mutations in these genes are the genetic basis of hemophilia A/B. To determine whether a sequence variation in F8/F9 is a disease-causing mutation, frequency data from a control population is needed. This study aimed to obtain data on sequence variation in F8/F9 in a set of functionally validated control chromosomes of Korean descent. METHODS: We re-sequenced F8 and F9 from DNA samples of 100 Korean male control individuals with normal PT, aPTT, and FVIII activity. PCR and direct sequencing analyses were performed using primer pairs to cover all coding regions and the flanking intronic sequences. RESULTS: Thirteen individuals (13%) were hemizygous for sequence variations in the coding region of F8. Six (6%) had c.3780C>G (p.Asp1260Glu), five (5%) had c.3864A>C (p.Ser1288=). One each individual (1%) had c.4794G>T (p.Glu1598Asp) and c.5069 A>G (p.Glu1690Gly). Asp1260Glu and Ser1288= were known SNPs (rs1800291 and rs1800292, respectively). Glu1598Asp was assigned as a missense mutation in public databases (HGMD and HAMSTeRS), and Glu1690Gly was a novel variation. Based on the normal FVIII activities in control individuals carrying these variations (109% and 148%, respectively), they were considered to be rare SNPs. No variation was observed in F9 of control individuals. CONCLUSION: A significant proportion of control individuals carried sequence variations in F8, but not in F9. These results can be used as a reference dataset for molecular diagnosis of hemophilia A and B, particularly in Korea.

Antiproliferation and Redifferentiation in Thyroid Cancer Cell Lines by Polyphenol Phytochemicals

Thyroid carcinogenesis is accompanied by loss of thyroid-specific functions and refractory to radioiodine and thyroid stimulating hormone (TSH) suppression therapy. Redifferentiating agents have been shown to inhibit tumor growth and improve the response to conventional therapy. Polyphenol phytochemicals (PPs) in fruits and vegetables have been reported to inhibit cancer initiation, promotion, progression and induce redifferentiation in selected types. In this study we examined PPs induce redifferentiation in thyroid cancer cell lines. We investigated the effects of genistein, resveratrol, quercetin, kaempferol, and resorcinol on the F9 embryonal carcinoma cell differentiation model. The thyroid cancer cell lines, TPC-1, FTC-133, NPA, FRO, and ARO, displayed growth inhibition in response to genistein, resveratrol, quercetin. We further demonstrated that genistein decreased the dedifferention marker CD97 in NPA cells and resveratrol decreased CD97 in FTC-133, NPA, FRO cells and quercetin decreased CD97 in all cell lines. We observed increased expression of differentiation marker NIS in FTC-133 cells in response to genistein, and resveratrol but no change in NPA, FRO, ARO cells. Quercetin increased or induced NIS in FTC-133, NPA, FRO cells. These findings suggest that PPs may provide a useful therapeutic intervention in thyroid cancer redifferentiation therapy.