ABSTRACT
Objective To investigate the clinical diagnostic value of circulating tumor cells (CTCs)and circulating cell-free DNA (cfDNA) in peripheral blood samples in breast cancer. Methods From July 2017 to April 2018, 47 patients with BMC (7 in stage Ⅱ, 19 in stage Ⅲ and 21 in stage Ⅳ), 24 patients with benign breast diseases and 28 healthy people were selected. After collecting peripheral blood samples, serum and blood cells were separated. The size-based high-throughput microfluidic chip was used to capture CTCs. The real-time fluorescent quantitative PCR based on Alu sequence was used to detect the length of cfDNA(247 bp, 115 bp)in the serum, and the ratio of amplified products of long and short fragments was used as the index of DNA integrity. The Mann-Whitney U test or Kruskal-Wallis H test was used to compare the differences between the groups and analyze the relationship between CTCs and cfDNA and clinical parameters of breast cancer. The ROC curve was drawn and the area under the curve (AUC) was used to evaluate the feasibility of blood cell CTCs and plasma cfDNA detection as diagnostic criteria. Results The CTCs and cfDNA of 47 BMC patients were analyzed. The CTCs and cfDNA integrity index (Alu 247/115) of BMC patients were significantly higher than those of physical examination patients[(13.98± 12.36)cells / ml vs (1.14 ± 1.35) cells / ml; 0.7687 ± 0.3868 vs 0.5094 ± 0.2456], and the difference was statistically significant(the U value was 126.5,359.0;P<0.001), the area under ROC curve of CTCs was 0.885 (95％CI: 0.805-0.965), cut-off value was 7.68/ml, sensitivity was 80.4％, specificity was 96.4％. The area under ROC curve of Alu 247/115 was 0.727(95％CI: 0.608-0.847), cut-off value was 0.431, sensitivity was 71.7％, specificity was 71.4％. The AUC of CTCs and Alu 247/115 was 0.919 (95％CI 0.854-0.984), which was higher than the single test of each indicator. Conclusions CTCs and cfDNA may be the potential biological indicators for breast cancer diagnosis. The combined detection of CTCs and cfDNA maybe improve the diagnosis rate of breast cancer patients.
ABSTRACT
Objective To investigate the clinical value of circulating miR-125b-5p in coronary atherosclerotic heart disease.Methods With case-control study,80 cases of coronary atherosclerotic heart disease were recruited in Affiliated Hospital of Nantong University from February 2014 to august 2015.According to coronary angiography result they were divided into two groups: there are coronary artery stenosis group(n=49)and control group(n=31).All patients were also divided into non-ST-segment elevation myocardial infarction and ST-segment elevation myocardial infarction group(n=35),unstable angina group group(n=25),stable angina group(n=20).The level of miR-125b-5p before coronary angiograph was detected.By independent sample t test and variance analysis,the levels of miR-125b-5p were compared between the groups of coronary artery stenosis and the group with no stenosis of the coronary artery,the coronary artery lesions in each group,and between the various types of coronary atherosclerotic heart disease respectively.Results MiR-125b-5p expression level of Coronary artery stenosis group(0.35±0.10)was lower than that in group coronary artery with no stenosis(0.95±0.12),the difference was statistically significant(t=24.179,P<0.000 1).With the increase in the number of diseased coronary arteries,miR-125b-5p expression level decreased gradually.There is also statistical significance(t=8.399,P<0.000 1; t=13.067,P<0.000 1)in miR-125b-5p expression among NSTEMI+STEMI,UA and SAP groups.miR-125b-5p expression level was negatively correlated with Gensini score(R2=0.822,P<0.05).The area under the ROC curve(AUC)of miR-125b-5p was 0.86(95％CI 0.67-0.90),and 0.66 was the optimal cut-off value with sensitivity of 81.22％and specificity of 78.62％.Conclusions With the increase of the number of stenosis,plasma miR-125b-5p expression level decreased gradually.The expression level of miR-125b-5p was negatively correlated with the Gensini score of coronary artery,which indicated that the expression level of miR-125b-5p may be a potential biomarker that can reflect the lesion degree of coronary artery.
ABSTRACT
Objective To investigate the role of APRIL in SW480 cell line.Methods The CRC model was established in the nude mice,all the mice were divided into 3 groups,the mice were separately treated with APRIL siRNA,pGC-vector and PBS solution.The APRIL mRNA was detected by RT-PCR and the APRIL protein was surveyed by the way of immunohis-tochemistry (IHC).The proteins of TIMP-3,Syndecan-1and MMP-9 also were assessed by IHC.Results ①Tumor mass in the group of nude mice injected with PBS (2.15±0.30 g)was significantly higher than the injection APRIL siRNA group (0.95±0.15 g,P0.05)compared with the injection of empty vector group (2.20±0.25 g).②APRIL mRNA/18S rRNA ratio (2.48±0.25)in the group of mice injected with PBS was signifi-cantly higher than the injection APRIL siRNA group (0.39±0.15,P0.05)compared with the injection of empty vector group (2.51±0.30).③SW480 cells injected with APRIL siRNA signifi-cantly inhibited invasion and metastasis.TIMP-3 Allred scores in three groups were 7.70±0.35,1.10±0.16 and 1.15± 0.12,Syndecan-1 protein was 7.80±0.30,1.05±0.20 and 1.10±0.22 MMP-9 protein was 1.20 ±0.10,8.00±0.25 and 8.20±0.20,respectively.Conlusion APRIL was closely connected with the growth and metabasis of CRC.