Expression level and clinical application value of human serum exosomal miR-218-5p in colorectal cancer / 中华检验医学杂志

Objective:To explore the expression level of exosomal miR-218-5p in the serum of patients with colorectal cancer (CRC) and its correlation with the clinical pathological characteristics, and evaluate its diagnostic efficacy in CRC.Methods:A group of 78 patients with colorectal cancer diagnosed in Tongji Hospital affiliated to Tongji University from October 2016 to October 2018 were selected. Blood was collected before operation and serum was preserved. Forty cases of healthy people in the same period were selected as the control group. ExoQuick kit was used to extract serum exosomes. Transmission electron microscope, NTA and western blot were used to identify the morphology and molecular phenotype of exosomes. MiRNeasy kit was used to extract total RNA in serum exosomes. Real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the expression level of serum exosomal miR-218-5p in each group. CRC patients were divided into high expression and low expression groups using the median relative expression level of miR-218-5p as the cut off value, and the four-square chi-square test (χ 2 test) was used to judge the relationship between miR-218-5p and clinicopathological features. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy of miR-218-5p in colorectal cancer. Results:The exosomes in serum were successfully extracted by kit method. The expression level of serum exosomal miR-218-5p in patients with colorectal cancer was significantly lower than that in normal healthy people [0.566(0.364, 0.850) vs 1.054(0.781, 1.709), P<0.001]. The low expression level was significantly better then the correlated with tumor size, TNM stage, lymph node metastasis and depth of invasion (all P<0.05). Receiver operating characteristic (ROC) analysis showed that the area under the curve (AUC) of serum exosomal miR-218-5p for the diagnosis of CRC was 0.827 (95% CI 0.754-0.900), which was significantly better than the conventional tumor marker carcinoembryonic antigen CEA (AUC =0.718, 95% CI 0.626-0.811) and carbohydrate antigen CA199 (AUC = 0.661, 95% CI 0.564-0.758). Conclusions:The down-regulation of miR-218-5p expression in serum exosomes of colorectal cancer patients is associated with a variety of adverse clinicopathological factors, which has the potential to become a diagnostic biomarker for colorectal cancer.

The application of free nucleic acid methylation in accurate diagnosis and treatment of tumor / 中华检验医学杂志

It is one of the most important forms of nucleic acid methylation in gene epigenetic modification and the most common driving molecular event in tumors. Detection of the free form nucleic acid can not only assist in early diagnosis and prognosis of tumors, but also avoid the problems of invasion and tumor heterogeneity caused by traditional tumor biopsy methods. In the present paper, the application status of free nucleic acid methylation detection in tumor diagnosis was reviewed, the challenges faced by free nucleic acid methylation detection were analyzed, and prospect of its clinical application was prospected.

Analyze the structure and component change of colonized bacterium in colorectal cancer tissues / 中华检验医学杂志

Objective To explore the structure and component changes of colonization bacterium in colorectal cancer (CRC) and analyze the relationship between the colonization bacteria and the development of CRC.Methods Clinical data of this retrospective study were obtained from Tongji Hospital of Tongji University (1/2016-10/2017).Forty matching tissues,including normal intestinal mucosa tissues (n=12) and CRC tissues (n=28),were collected in the present study.The V3-V4 region of bacteria 16S rRNA gene was detected using Illumina Miseq sequencing technologies.The colonized bacterium structure and composition were analyzed and compared between the two groups.Results It was found that there is significant difference in structure of the colonized bacterium between the two groups.In the level of Phylum,the abundance of Firmicutes and Bacteroidetes were significant lower (Z=-1.964,P＜0.05),and Proteobacteria was significant higher (Z=-1.997,P＜0.001) in the tissues of CRC.In the level of genus,Bacteroides,Blautia,Prevotella,and Faecalibacterium were significant lower in the tissues of CRC (Z=-3.008,P＜0.05).Buchnera,Prevotella,and Prevotellaceae only existed in normal intestinal mucosa tissues and the abundance was top three.Catonella,Kurthia,and Brachymonas only existed in CRC tissues and the abundance was top three.Conclusions The overall structure and component were significant difference between the two groups.Some bacterium disappeared or reduced,and others new emerged or increased.The change of colonized bacterium structure and component in intestinal mucosa may play an important role in the development of CRC.Therefore,it may be an,accurate and low cost of early diagnosis marker by identifing the structure and component change of intestinal mucosa colonized bacterium for CRC,which may find a new way for prevention and treatment of CRC.

The clinical application value of human serum exosomal CEA in the diagnosis of colorectal cancer / 中华检验医学杂志

Objective@#To isolate and identify exosomes from human serum, explore the feasibility of exosomal CEA for the diagnosis of colorectal cancer.@*Methods@#Retrospective study.64 cases with colorectal cancer patients(41 cases with normal CEA results and 23cases with high CEA results), 20 cases with benign colorectal diseases patients and 40 cases with healthy people of department of physical examination from October 2015 to December 2016 in Tongji Hospital of Tongji University. Exosomes were isolated from these serum using ExoQuick, and then identified by using transmission electron microscopy, and Western Blot for morphology and molecular phenotype.The serum level of CEA and exosomal CEA was measureed by enzyme linked immunosorbent assay (ELISA). The diagnostic efficacy of serum Exosomal CEA concentration in the colorectal cancer by using U test and the subject work characteristic curve (ROC). @*Results@#Exosomes in human serum was successfully extracted with ExoQuick kit.Exosomal CEA concentration in 64 cases with colorectal cancer patients (1 056.36-28 637.78)pg/ml was significantly higher than in cases with benign colorectal diseases patients (394.61-2 437.83)pg/ml and healthy controls(610.89-2 076.70)pg/ml(U=124.000, U=119.000, P<0.01). Exosomal CEA concentration in 41 colorectal cancer patients with normal serum CEA concentration(1 056.36-5 832.07)pg/ml was significantly higher than in benign colorectal diseases patients and healthy controls(U=113.000, U=119.000; P<0.01). ROC analysis of exosomal CEA yielded an AUC(area under the ROC curve)of 0.954(95% CI=0.919-0.988), which was higher than the serum CEA.The area under the ROC curve of serum Exosomal CEA concentration for colorectal cancer diagnosis is 0.954(95% CI=0.919-0.988), which is superior to the serum CEA concentration[0.636 (95% CI=0.531-0.742)]. @*Conclusions@#Isolation and detection of serum Exosomal CEA concentrations have a good diagnostic efficacy in colorectal cancer. It′s possible to be a marker for early diagnosis of colorectal cancer.(Chin J Lab Med, 2018, 41: 503-508)

Antimicrobial peptide LL-37 in macrophages promotes colorectal cancer growth / 中华肿瘤杂志

Objective@#To investigate the effect and molecular mechanism of antimicrobial peptide LL-37 secreted by stromal cells on the growth of colorectal cancer cells.@*Methods@#Colorectal cancer cells SW480 or HCT116 were co-cultured with human macrophages using Transwell® maxicell inserts to mimic the tumor microenvironment. The effect of macrophages on the proliferation of colorectal cancer cells was detected by Bromodeoxyuridine and enzyme-linked immunosorbent assay (BrdU-ELISA). The expression of LL-37 mRNA and protein in macrophages and colorectal cancer cells was evaluated by reverse transcription-real-time quantitative PCR (RT-qPCR) and Western blot. LL-37 neutralizing antibody was added to abrogate the LL-37 activation. Additionally, macrophages were transfected with LL-37 shRNA plasmids to inhibit LL-37 expression. And then, the proliferation of colorectal cancer cells was observed. Furthermore, the growth-related signaling pathways were detected by Western blot.@*Results@#The BrdU-ELISA results showed that the absorbance of SW480 cells increased from 1.072±0.097 to 5.121±0.407 after co-culture (P<0.001), and that of HCT116 cells increased from 1.229±0.073 to 3.495±0.228 (P<0.001). RT-qPCR results showed that LL-37 mRNA expression in macrophages significantly increased from 2.682±0.191 to 6.117±0.768 after co-incubation (P<0.05), whereas that in SW480 had no significant difference. Consistently the protein expression of LL-37 in macrophages was significantly increased by Western blot, while it did not change in SW480. The proliferation rate of SW480 cells was repressed by adding LL-37 neutralizing antibody or LL-37 shRNA plasmid. Furthermore, Western blot analysis showed that the expression of non-phosphorylated (activated) β-catenin and its target genes cyclin D1 as well as c-myc were distinctly increased in co-cultured SW480 cells, which could be reversed by anti-LL-37 antibodies.@*Conclusion@#Macrophages promote the in vitro proliferation of colorectal cancer cells by enhancing the expression and secretion of antimicrobial peptides LL-37, and it seems that LL-37 activates colorectal cancer cells via Wnt/β-catenin pathway.

Auxiliary diagnosis value of serum cytokines and neurotrophic factor levels in schizophrenia inpatiens / 国际检验医学杂志

Objective To investigate the changes of serum IL-1β,IL-6 ,TNF-α,NGF and BDNF levels and their correlation with clinical symptoms of schizophrenia inpatiens and their value in the auxiliary diagnosis of schizophrenia .Methods The case-control study was used .The levels of serum IL-1β,IL-6 ,TNF-α,NGF and BDNF were measured by using the enzyme linked immunosorbent assay(ELISA) in 85 inpatients with schizophrenia and 85 healthy controls .Their changes in the case group were compared between before treatment and after 3-month treatment .The Pearson correlation analysis was used to analyze the correlation between the lev-els of IL-1β,IL-6 ,TNF-α,NGF and BDNF with the positive and negative syndrome scale (PANSS) score and the auxiliary diagnosis value of serum cytokine and neurotrophic factor levels were evaluated with the receiver operating characteristic (ROC) curve .Re-sults The levels of serum IL-1β(t=4 .560) ,IL-6(t= 4 .957) and TNF-α(t= 4 .799) before treatment in the schizophrenia case group were significantly higher than those in control group ,while the NGF(t= -4 .806) and BDNF(t= -4 .881) levels were sig-nificantly lower than those in the control group ,the differences were statistically significant (P<0 .01) .After 3-month treatment , the levels of serum IL-1β(t=4 .543) ,IL-6(t=4 .327) and TNF-α(t=4 .654) in the schizophrenia case group were significantly de-creased compared with before treatment ,while the NGF(t= -4 .641) and BDNF(t= -4 .876) levels were significantly increased , the differences were statistically significant (P< 0 .01) .IL-1β was positively correlated with the positive symptoms scores (r=0 .325 ,P<0 .01) ,IL-6 was positively correlated with the negative symptoms scores (r=0 .319 ,P<0 .01) ,TNF-α was positively correlated with the positive symptoms scores (r= 0 .281 ,P< 0 .01) ,NGF was negatively correlated with the positive symptoms scores(r= -0 .229 ,P<0 .05) ,BDNF was negatively correlated with the positive symptoms scores (r= -0 .272 ,P< 0 .05) .The cut-off values of serum IL-1β,IL-6 ,TNF-α,NGF and BDNF in the auxiliary diagnosis of schizophrenia were 40 .083 ,20 .037 ,17 .115 ,19 .998 ,584 .157pg/mL respectively ,the corresponding areas under the ROC were 0 .723 ,0 .772 ,0 .686 ,0 .712 and 0 .708 respectively ,the sensitivities were 0 .565 ,0 .871 ,0 .894 ,0 .859 and 0 .729 respectively ,and the specificities were 0 .871 ,0 .565 ,0 . 365 ,0 .494 and 0 .624 respectively .Conclusion The levels of serum IL-1 ,IL-6 ,TNF-α,NGF and BDNF have the correlation with the clinical symptoms of schizophrenic inpatients and have a certain value in the auxiliary diagnosis of schizophrenia .

Influence of oxidative stress on p53 post-translational modifications / 上海交通大学学报（医学版）

Protein post-translational modifications are important ways to regulate protein function and cell behavior, and oxidative stress directly influences protein post-translational modifications. P53 protein has various post-translational modifications, which can be quickly regulated under stress conditions to activate a series of downstream target genes and facilitate the p53 function diversity. The effects of oxidative stress on p53 post-translational modifications of phosphorylation, ubiquitination, sumoylation, acetylation and methylation are introduced in this paper.