Identification of potential diagnostic and prognostic biomarkers for breast cancer based on gene expression omnibus.

BACKGROUND: Breast cancer is regarded as a highly malignant neoplasm in the female population, posing a significant risk to women's overall well-being. The prevalence of breast cancer has been observed to rise in China, accompanied by an earlier age of onset when compared to Western countries. Breast cancer continues to be a prominent contributor to cancer-related mortality and morbidity among women, primarily due to its limited responsiveness to conventional treatment modalities. The diagnostic process is challenging due to the presence of non-specific clinical manifestations and the suboptimal precision of conventional diagnostic tests. There is a prevailing uncertainty regarding the most effective screening method and target populations, as well as the specificities and execution of screening programs. AIM: To identify diagnostic and prognostic biomarkers for breast cancer. METHODS: Overlapping differentially expressed genes were screened based on Gene Expression Omnibus (GSE36765, GSE10810, and GSE20086) and The Cancer Genome Atlas datasets. A protein-protein interaction network was applied to excavate the hub genes among these differentially expressed genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses, as well as gene set enrichment analyses, were conducted to examine the functions of these genes and their potential mechanisms in the development of breast cancer. For clarification of the diagnostic and prognostic roles of these genes, Kaplan-Meier and Cox proportional hazards analyses were conducted. RESULTS: This study demonstrated that calreticulin, heat shock protein family B member 1, insulin-like growth Factor 1, interleukin-1 receptor 1, Krüppel-like factor 4, suppressor of cytokine signaling 3, and triosephosphate isomerase 1 are potential diagnostic biomarkers of breast cancer as well as potential treatment targets with clinical implications. CONCLUSION: The screening of biomarkers is of guiding significance for the diagnosis and prognosis of the diseases.

A novel ratiometric fluorescent probe based on dual-emission carbon dots for highly sensitive detection of salicylic acid.

In this study, a novel ratiometric fluorescence probe based on dual-emission carbon dots (CDs) for the sensitive detection of salicylic acid (SA) was constructed for the first time. The dual-emission CDs were synthesized by simple hydrothermal method using tartaric acid (TA) and m-phenylenediamine (mPD) as raw materials. In the presence of SA, the fluorescence intensity of CDs was enhanced at 499 nm, but remained basically unchanged at 439 nm. This phenomenon is caused by the intermolecular hydrogen bond interactions. The concentrations of SA had an excellent linear relationship with CDs' fluorescence intensity ratio (F499/F439) in a range of 1 âˆ¼ 120 and 120 âˆ¼ 240 µM with low detection limits of 0.68 and 1.05 µM. The established ratiometric fluorescent probe is economical, simple and green, and can be used for the effective detection of SA. In addition, the proposed ratiometric fluorescent probe was successfully used to monitor SA in facial mask and toning lotion samples with a satisfactory recovery of 99.7-106.7 %. The results show that the constructed fluorescent probe based on dual-emission CDs has a great potential for the rapid and sensitive analysis of SA in actual samples.

[Hydroxysafflor yellow A inhibits proliferation, migration, and chemoresistance of colorectal cancer cells through Akt/mTOR-autophagy pathway].

In recent years, the clinical treatment of colorectal cancer(CRC) has made great progress, but chemoresistance is still one of the main reasons for reducing the survival rate of patients with colorectal cancer. Therefore, ameliorating chemotherapy resis-tance is an urgent problem to be solved. The purpose of this study was to investigate the regulatory role and related molecular mechanisms of hydroxysafflor yellow A(HSYA) in colorectal cancer cell proliferation, migration, and 5-fluorouracil(5-FU) chemoresistance. In this study, HCT116 and HT-29 cells were used as research subjects. Firstly, methyl thiazolyl tetrazolium(MTT) assay and colony formation assay were used to detect and analyze the effect of HSYA on the proliferation of CRC cells. Secondly, the effect of HSYA on the cell cycle in CRC cells was analyzed by cell cycle assay. Furthermore, the effect of HSYA on the migration of CRC cells was analyzed by wound-healing assay and Transwell assay. Based on the above, the influences of HSYA on 5-FU chemoresistance of CRC cells and related molecular mechanisms were explored and analyzed. The results showed that HSYA significantly inhibited the proliferation and migration of CRC cells, and arrested the cell cycle in G_0/G_1 phase. In addition, HSYA significantly ameliorated the chemoresistance of CRC cells to 5-FU. The results of acridine orange staining and Western blot showed that the autophagy activity of CRC cells in the HSYA and 5-FU combined treatment group was significantly higher than that in the 5-FU single drug treatment group. As compared with the 5-FU single drug treatment group, the phosphorylation levels of protein kinase B(Akt) and mammalian target of rapamycin(mTOR) in the HSYA and 5-FU combined treatment group were significantly reduced, indicating that the Akt/mTOR signaling pathway in the combined treatment group was down-regulated in CRC cells. In conclusion, HSYA may upregulate autophagy activity through the Akt/mTOR signaling pathway, thereby inhibiting the proliferation and migration of CRC cells and ameliorating the chemoresistance to 5-FU.

Selective colorimetric and fluorescence detection of nitroreductase enzymes in living cells.

Rhodamine dyes bearing aromatic nitro group has been synthesized for nitroreductase enzyme chemosensing applications. The probe is showing very selective turn-on fluorescent response towards nitroreductase enzymes and in hypoxic conditions. The sensor displays a remarkable fluorescent enhancement at 557 nm (λex = 500 nm) without the interference of other biologically relevant species under hypoxic conditions in a physiological medium. The nitro group in the sensor is reduced by the nitroreductase enzyme to the amino group, resulting in the hydrolysis of the probe and subsequent release of highly fluorescent rhodamine 6G dye is observed. This rhodamine based fluorescent probe has been utilized for the imaging of nitroreductase enzymes as well as hypoxia in live cells.

Comparison of single- and triple-injection methods for ultrasound-guided interscalene brachial plexus blockade.

Ultrasound-guided interscalene brachial plexus blockade (IBPB) has a relatively high success rate in shoulder surgery; however, whether multiple injections are superior to a single injection (SI) is currently unknown. In the present study, ultrasound-guided SI and triple-injection (TI) IBPBs were compared in a prospective randomized trial. A total of 111 patients undergoing arthroscopic shoulder surgery and presenting with an American Society of Anesthesiologists physical status grading of I-II were randomly allocated to receive IBPB with 15 ml of 1% ropivacaine as a SI or TI. Performance time, procedure-related pain scores, success rate and prevalence of complications were recorded. The distribution of sensory and motor block onset in the radial, median, ulnar and axillary nerves were assessed every 5 min until 30 min post-local anesthetic injection. The duration of sensory and motor blocks were also assessed. A significantly longer performance time was recorded in the TI group (P<0.001). No significant difference was observed in success rate (91% in TI vs. 88% in SI) 30 min post-injection, and the prevalence of complications and procedure-related pain were similar between the two groups. Sensory and motor blocks of the ulnar nerve in the TI group were significantly faster and more successful compared with the SI group at all time points (P<0.041). It was also observed that sensory and motor blocks in the TI group were prolonged compared with the SI group (P<0.041). In conclusion, the TI method exhibited a faster time of onset and resulted in a more successful blockade of the ulnar nerve. TI method may be a more effective approach for IBPB in a clinical setting.

[Construction of HCV-producing cell model based on self-cleaving ribozyme].

OBJECTIVES: To construct a stable HCV-producing cell model for anti-HCV drug research. METHODS: The HCV-ribozyme recombinant plasmid pJFH1-Rbz was constructed to generate the exact 5' and 3' ends of HCV genomic RNA by placing two self-cleaving ribozymes at both ends of the HCV JFH-1 cDNA. The plasmid was then transfected into HepG2 cells and the resultant clones were screened with G418. Subsequently, immunofluorescence and Western blot were performed to detect the expression of HCV core protein, HCV RNA level was quantitated by TaqMan real-time PCR method and HCV particles was detected by electron microscopy. RESULTS: HCV core protein was detected in the screened cell clone, and the level of HCV RNA was up to 1000,0000 copies/ml in the culture medium. Electron microscopy showed the viral particles in the culture suspension were approximately 55 nm in diameter. IFN-treating experiment demonstrated that the HCV RNA level decreased with the increasing concentration of IFN alpha. CONCLUSION: We constructed a stable HCV-producing cell model which can be used for anti-HCV drug research.

[Construction of the nucleic vaccine pVVP3L-18HN and its antitumor effect on human laryngeal carcinoma].

OBJECTIVE: Nucleic vaccine of pVVP3IL-18HN expressing apoptin gene, Newcastle disease virus HN gene and IL-18 gene were constructed to observe the combinative antitumor effect of the above three genes. METHODS: Eukaryotic expression plasmid pVVP3IL-18HN was constructed by inserting apoptin gene and fragment comprising fused IL-18HN gene and IRES promoter into the downstream of CMV promoter of vector pVAX1. The expression of inserted gene was identified by RT-PCR, indirect immunofluorescence and Western-blot. The recombinant plasmid was introduced into Hep-2 cells by liposome, then suppression rate of Hep-2 of different time and different quantity was calculated according to MTT results. RESULT: The recombinant plasmid of pVVP3IL-18HN suppressed Hep-2 successfully and its suppression rate was up to 61.9% with 20 microg/ml, incubation of 72 hours. CONCLUSION: The nucleic vaccine constructed pVVP3IL-18HN had antitumor effect on Hep-2. It may can be used to the therapy and research of laryngeal carcinoma.

[Induction of apoptosis in human hepatoma cell line SMMC7721 by Newcastle disease virus HN gene].

OBJECTIVE: To investigate the mechanisms of apoptosis induced in human hepatoma cell line SMMC7721 by plasmid pVHN constructed with Newcastle disease virus (NDV) HN gene. METHODS: Twenty-four h after transfection with liposome-plasmid pVHN complexes in vitro, the mortality rate of SMMC7721 cells was determined by MTT staining and flow cytometry (FCM) with PI staining. The alteration of mitochondrial trans-membrane potential of the cells was detected by FCM with rhodamine 123 staining. Cell genomic DNA was detected by agarose electrophoresis. The activation of caspase-3 was assayed by its substrate color reaction. RESULTS: Significant apoptosis was induced by transfection with plasmid pVHN into the cells for 24 h and the mortality rate was 50.0% (the mortality rate of control group was 5.2%). Genomic DNA was fragmented and mitochondrial trans-membrane potential was decreased, but caspase-3 activity increased. CONCLUSION: Significant apoptosis in SMMC7721 cells can be induced by NDV HN gene. Apoptosis may be resulted from the decrease of mitochondrial trans-membrane potential and activation of Caspase-3.

[Antitumor research on mouse melanoma with combined application of Newcastle disease virus and its HN gene].

BACKGROUND & OBJECTIVE: Although Newcastle disease virus (NDV) shows antitumor effect on many tumors, its mechanism is unclear. Hemagglutinin-neuraminidase (HN) gene was found to play an important role in NDV antitumor effect and HN protein located on tumor cell surface. This research was to evaluate the possibility of HN protein as a foreign antigen of tumor cell and the antitumor effect of the combined application of HN gene and NDV. METHODS: C57BL/6 mice were subcutaneously inoculated with 2 x 10(5) B16 tumor cells in the right hindlimb. Combination group: on 2nd day post-inoculation, the recombinant plasmid containing HN gene was injected intramuscularly in the left hindlimb; on 7th day post-inoculation, 2 x 10(9) pfu NDV was administrated intratumorally. The alone HN gene group, NDV group, and PBS control group were treated as above. The antitumor effect was observed through tumor suppression rate, the antitumor mechanisms were researched with specific cytotoxic T lymphocyte (CTL) assay, and the expression determination of HN protein, ICAM-I, and CD48 on the B16 tumor cells. RESULTS: The antitumor efficacy of the combined application of NDV and its HN gene increased compared with NDV,and its HN gene alone, the tumor suppression rates were 82.8%, 41.0%, and 56.6%; the specific CTL activity were 18.4%, 10.1%, and 4.4%, respectively. Furthermore, the expression of HN gene had been detected, and the expression of ICAM-I and CD48 were up-regulated on the tumor cells after NDV injection. CONCLUSION: HN protein located on the surface of tumor cells and mediated the specific repulsion to tumor cells; the antitumor efficacy increased after the combined application of NDV and its HN gene.