目的 探讨外周血血管内皮生长因子(VEGF)、循环肿瘤细胞(CTCs)与乳腺癌新辅助化疗疗效的相关性.方法 选取沈阳市中国医科大学附属盛京医院乳腺外科2018年2月-2019年2月行新辅助治疗的30例乳腺癌患者为研究对象,根据Recist疗效评价标准分为4组,CR组(4例),PR组(20例),SD组(6例),PD组(0例),分别于入院时、化疗6周期后检测外周血VEGF、CTCs水平,采用配对t检验对各组检测指标进行分析,采用Pearson分析对新辅助化疗前后VEGF及CTCs水平之差进行分析.结果 新辅助化疗有效组(CR、PR组)化疗前血清VEGF水平分别为(324.16±26.11)pg/mL和(237.95±109.56)pg/mL,化疗6周期后降为(197.96±59.99)pg/mL和(153.98±82.57)pg/mL,与化疗前相比差异有统计学意义(t=5.287、8.684,P＜0.05);新辅助化疗无效组(PD、SD组),其中PD组为0例,SD组化疗前血清VEGF水平为(213.06±112.77)pg/mL,化疗6周期后降为(131.28±81.81)pg/mL,与化疗前相比差异有统计学意义(t=5.837,P=0.002);有效组(CR、PR组)化疗前CTCs水平分别为(4.25±0.83)个/3.2 mL 和(5.90±1.37)个/3.2 mL,化疗6周期后降为(1.25±0.43)个/3.2 mL 和(3.15±1.62)个/3.2 mL,与化疗前相比差异有统计学意义(t=5.623、9.491,P＜0.05);SD组化疗前后CTCs水平相比未见明显变化,差异无统计学意义(t=1.348,P=0.235);新辅助化疗前后VEGF水平变化与CTCs水平变化相关(|S|=0.636,P＜0.05).结论 VEGF水平与CTCs水平同新辅助化疗疗效密切相关,两者的变化在新辅助化疗过程中呈正相关,联合两者对新辅助疗效进行评估有重大价值.

随着对乳腺癌发病机制及疾病进展的深入研究,新辅助化疗已然成为炎性乳腺癌,局部晚期乳腺癌和选择保乳手术治疗患者的首选治疗措施.因此,新辅助化疗的疗效评估成为治疗的关键环节.临床上大多以影像学资料作为首选方法,血清学指标的应用少之又少.循环肿瘤细胞(CTCs)作为一项日趋成熟的血清学标志物,具有取材方便,检测手段易行,机体创伤小,可重复监测等优点,与传统影像学资料相比,CTCs能够较早发现微转移,无射线损伤等优势,应用前景广泛.本文就CTCs在乳腺癌新辅助化疗疗效评估中的价值作一综述.

乳腺癌为女性最常见的恶性肿瘤,发病率和致死率逐年上升,且有年轻化趋势[1].随着对乳腺癌发病机制及疾病进展的深入研究,发现乳腺癌是一种血管依赖型恶性肿瘤,其血行转移和淋巴转移都与血管的过度形成密不可分,而血管的过度形成是多个促血管形成因子共同作用的结果.血管内皮生长因子(vascular endothelial growth factor,VEGF)为目前已知的作用最强的血管生长刺激因子,探究其在乳腺癌组织和血清中的表达及作用机制有重大的临床意义.

Breast cancer (BC) is a leading cause of cancer mortality in women worldwide. MAC30/Transmembrane protein 97 (TMEM97) is aberrantly up-regulated in many human carcinoma cells. However, the function of MAC30 in invasion and EMT of BC cells is uncertain. qRT-PCR was used to determine the level of MAC30 in BC tissues and cell lines. si-MAC30 was transfected into BC cells, and the effects of MAC30 silencing on the invasion and EMT were explored by qRT-PCR as well as transwell and western blot assays. Also, we determined the effects of MAC30 silencing on Wnt/β-catenin and PI3K/Akt signaling pathways by western blot. We found that MAC30 is significantly up-regulated in BC tissues and cell lines. Down-regulation of MAC30 expression efficiently inhibited the invasion of BC cells. Furthermore, the EMT of BC cells was also inhibited by down-regulation of MAC30. Finally, we found that MAC30 knockdown inhibited Akt phosphorylation, β-catenin, survivin, and cyclin D1 expressions. To our knowledge, this is the first report investigating the effect of MAC30 on invasion and EMT in BC cells by suppressing Wnt/β-catenin and PI3K/Akt signaling pathways. MAC30 may be a potential therapeutic target for BC.

Breast cancer (BC) is a leading cause of cancer mortality in women worldwide. MAC30/Transmembrane protein 97 (TMEM97) is aberrantly up-regulated in many human carcinoma cells. However, the function of MAC30 in invasion and EMT of BC cells is uncertain. qRT-PCR was used to determine the level of MAC30 in BC tissues and cell lines. si-MAC30 was transfected into BC cells, and the effects of MAC30 silencing on the invasion and EMT were explored by qRT-PCR as well as transwell and western blot assays. Also, we determined the effects of MAC30 silencing on Wnt/β-catenin and PI3K/Akt signaling pathways by western blot. We found that MAC30 is significantly up-regulated in BC tissues and cell lines. Down-regulation of MAC30 expression efficiently inhibited the invasion of BC cells. Furthermore, the EMT of BC cells was also inhibited by down-regulation of MAC30. Finally, we found that MAC30 knockdown inhibited Akt phosphorylation, β-catenin, survivin, and cyclin D1 expressions. To our knowledge, this is the first report investigating the effect of MAC30 on invasion and EMT in BC cells by suppressing Wnt/βcatenin and PI3K/Akt signaling pathways. MAC30 may be a potential therapeutic target for BC.

BACKGROUND:Breast cancer, the most common invasive cancer of women, is a malignant neoplasm and the second main cause of cancer death. Resistance to paclitaxel (Taxol), one of the frequently used chemotherapy agents for breast cancer, presents a major clinical challenge. Recent studies revealed that metabolic alterations of cancer cells play important roles in chemo-resistance.MATERIALS AND METHODS:In this study, Human breast cancer cells, BT474, SKBR3 and MCF7 were used to study the causal relationship between the lactate exporter, MCT1 (SLC16A1)-modulated glucose metabolism and Taxol resistance of breast cancer cells. Taxol resistant breast cancer cells were established. The intracellular lactate and extracellular lactate levels as well glucose uptake and oxygen consumption were measured. MicroRNA-124 expressions were detected by qRT-PCR from both breast cancer patient samples and breast cancer cells. Target of miR-124 was predicted and verified by Western blot and luciferase assay. An xenograft mice model was established and evaluated for the in vivo tumor therapeutic effects of MCT1 inhibitor plus microRNA-124 treatments.RESULTS:Low toxic Taxol treatments promoted cellular glucose metabolism and intracellular lactate accumulation with upregulated lactate dehydrogenase-A (LDHA) and MCT1 expressions. By establishing Taxol resistant breast cancer cell line, we found Taxol resistant cells exhibit upregulated LDHA and MCT1 expressions. Furthermore, glucose consumption, lactate production and intracellular ATP were elevated in Taxol resistant MCF7 cells compared with their parental cells. The miR-124, a tumor suppressive miRNA, was significantly downregulated in Taxol resistant cells. Luciferase assay and q-RT-PCR showed MCT1 is a direct target of miR-124 in both breast cancer cell lines and patient specimens. Moreover, co-treatment of breast cancer cells with either MCT1 inhibitor or miR-124 plus Taxol led to synergistically cytotoxic effects. Importantly, based on in vitro and in vivo results, inhibition of MCT1 significantly sensitized Taxol resistant cells. Finally, rescue experiments showed restoration of MCT1 in miR-124 overexpressing cells promoted Taxol resistance.CONCLUSIONS:This study reveals a possible role of miRNA-214-mediated Taxol resistance, contributing to identify novel therapeutic targets against chemoresistant breast cancers.

Background: L-proline is a natural, nontoxic cryoprotectant that helps cells and tissues to tolerate freezing in a variety of plants and animals. The use of L-proline in mammalian oocyte cryopreservation is rare. In this study, we explored the cryobiological characteristics of L-proline and evaluated its protective effect in mouse oocyte cryopreservation.Methods: The freezing property of L-proline was detected by Raman spectroscopy and osmometer. Mature oocytes obtained from 8-week-old B6D2F1 mice were vitrified in a solution consisting various concentration of L-proline with a reduced proportion of dimethyl sulfoxide (DMSO) and ethylene glycol (EG), comparing with the control group (15 % DMSO and 15% EG without L-proline). The survival rate, 5-methylcytosine (5-mC) expression, fertilization rate, two-cell rate, and blastocyst rate in vitro were assessed by immunofluorescence and in vitro fertilization. Data were analyzed by Chi-square test.Results: L-proline can penetrate the oocyte membrane within 1 min. The osmotic pressure of 2.00 mol/L L-proline mixture is similar to that of the control group. The survival rate of the postthawed oocyte in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG is significantly higher than that of the control group. There is no difference of 5-mC expression between the L-proline combination groups and control. The fertilization rate, two-cell rate, and blastocyst rate in vitro from oocyte vitrified in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG solution are similar to that of control.Conclusions: It indicated that an appropriate concentration of L-proline can improve the cryopreservation efficiency of mouse oocytes with low concentrations of DMSO and EG, which may be applicable to human oocyte vitrification.

Recent studies have shown that L-proline is a natural osmoprotectant and an antioxidant to protect cells from injuries such as that caused by freezing and thawing in many species including plant, ram sperm and human endothelial cells. Nevertheless, this nontoxic cryoprotectant has not yet been applied to mammalian oocyte vitrification. In this study we evaluated the efficiency and safety of the new cryoprotectant in oocyte vitrification. The results indicated that L-proline improves the survival rate of vitrified oocytes, protects mitochondrial functions and could be applied as a new cryoprotectant in mouse oocyte vitrification.

Objective To investigate the expression profile of mRNAs in brain samples collected from pronuclear transfer (PNT) mice. Methods Female CD-1 mice were superovulated, and zygotes were collected after mating with adult male mice. Zygotes with two pronuclei were selected for pronuclear transfer manipulation, and then the reconstructed zygotes were transferred into the oviduct of pseudopregnant female mice. The infant mice obtained from pronuclear transfer were called PNT group, while the embryoes that were not performed pronuclear transfer was regarded as control group. Total RNA were extracted from brain samples of both PNT and control mice, and cDNA were labeled with fluorescent dye. Genes that were differentially expressed were identified using the Agilent mouse mRNA array. Gene ontology analysis and pathway analysis were also completed. Results Compared with control group, 392 mRNAs were expressed differentially, which showed more than 2.0 times variation and statistical significance, accounting for 1.7% of all mRNAs. Among those 366 mRNAs were up-regulated and 26 mRNAs were down-regulated. Eleven mRNAs came to 4.0 times variation in total. Gene ontology analysis indicated that differentially expressed genes were significantly enriched in alternative mRNA splicing, small GTPase mediated signal transduction, regulation of insulin receptor signaling pathway, hydrolase activity, transmembrane transporter activity and pyrophosphatase activity. Significant enriched pathway terms contained ion channel transport, fatty acid metabolism, butanoate metabolism, triacylglycerol and ketone body metabolism. Conclusion Pronuclear transfer might influence some key metabolism process in mouse brain.

OBJECTIVE To determine the diagnostic features of Robertsonian (Rob) translocation (11; 13) in mice and the mechanisms underlying the effect on spermatogenesis and reproductive decline. METHODS A Rob translocation (11; 13) mouse model was established by cross-breeding, and confirmed by chromosome analysis. Chromosome aberrations and translocation patterns were identified in mice with Rob translocation (11; 13) by fluorescence in situ hybridization (FISH). Spermatogenic disorders were investigated at different stages of spermatogenesis. Immunofluorescent analysis was performed on sections of testis and epididymis specimens during spermatogenic meiosis. The weight of the testes and reproductive decline were recorded. RESULTS The crossed Rob translocation (11; 13) mouse has 39 chromosomes, including a fusion chromosome (included chromosomes 11 and 13) using dual color FISH. There was no difference in the distribution pattern of SYCP3 and γH2AX in spermatocytes between Rob translocation and wild-type mice; however, round haploid spermatids presented characteristic morphologic changes of apoptosis and the number of haploid spermatids was decreased. Furthermore, the immature germ cells were released into the epididymis and the number of mature sperm was reduced. CONCLUSIONS Chromosome aberrations and spermatogenic disorders may result from apoptosis of round haploid spermatids and a reduced number of mature sperm in Rob translocation (11; 13) mice. Abnormal sperm and reduced number of sperm may be one of the main reasons for reproductive decline and male infertility in Rob translocation (11; 13) mice.

DNA methylation is a crucial element in the epigenetic regulation of mammalian embryonic development. However, its dynamic patterns have not been analysed at the genome scale in human pre-implantation embryos due to technical difficulties and the scarcity of required materials. Here we systematically profile the methylome of human early embryos from the zygotic stage through to post-implantation by reduced representation bisulphite sequencing and whole-genome bisulphite sequencing. We show that the major wave of genome-wide demethylation is complete at the 2-cell stage, contrary to previous observations in mice. Moreover, the demethylation of the paternal genome is much faster than that of the maternal genome, and by the end of the zygotic stage the genome-wide methylation level in male pronuclei is already lower than that in female pronuclei. The inverse correlation between promoter methylation and gene expression gradually strengthens during early embryonic development, reaching its peak at the post-implantation stage. Furthermore, we show that active genes, with the trimethylation of histone H3 at lysine 4 (H3K4me3) mark at the promoter regions in pluripotent human embryonic stem cells, are essentially devoid of DNA methylation in both mature gametes and throughout pre-implantation development. Finally, we also show that long interspersed nuclear elements or short interspersed nuclear elements that are evolutionarily young are demethylated to a milder extent compared to older elements in the same family and have higher abundance of transcripts, indicating that early embryos tend to retain higher residual methylation at the evolutionarily younger and more active transposable elements. Our work provides insights into the critical features of the methylome of human early embryos, as well as its functional relation to the regulation of gene expression and the repression of transposable elements.

The failure of anticorrosion turn up in many chimneys as the retrofitting project of wet FGD for thermal power plant finished.Anticorrosion retrofitting must be carried out to eliminate the risk taken by the corrosion of chimney.Temporary stacks will be installed to make sure the plant operating normally.So the design is introduced,so as to offer for reference in other similar projects.

Applications of fly ash discharged from coal-fired power plant in Chinese architectural industry include road building materials,dam base or slab foundation,raw material to produce cement,an admixture for concrete or mortar,manufacturing building blocks or building elements and other building products in alliance with plastic scraps.Measures of enhancing the quality of Fly ash discharged from coal-fired power plant are as follows:ameliorating conditions for coal dust to sufficiently burn to raise the efficiency of combustion,carrying out strict size classification of coal dust by its particle properties and physical and chemical properties.

Based on the characteristics of Handan electric power plant No.5 construction,combined with soil parameter and the experience of No.4 construction,the reason which brought on the upfloat using pile foundation,and raises the integrated countermeasures,the result is perfect,and satisfies the design need.The experiences may give full assistance on construction,designing,and field survey for reference.