Objective:To evaluate the diagnostic efficacy of plasma exosomal lncNORAD as a tumor marker for bladder cancer (BCa) by examining its expression levels in BCa patients and analyzing the correlation with clinical pathological characteristics.Methods:This retrospective study. 100 BCa patients diagnosed between October 1, 2021, and October 1, 2023, at Tongji Hospital, Tongji University were randomly selected as the patient group (76 males, 24 females, mean age 71±10 years). In the same period, 40 patients with benign urinary system diseases (benign disease group; 33 males, 7 females, mean age 64±9 years) and 70 healthy individuals undergoing routine check-ups (control group; 53 males, 17 females, mean age 45±12 years) were included. Plasma exosomes were extracted using an exosome extraction kit, and morphologically and molecularly were characterized by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting. Total RNA from plasma exosomes was extracted, and the expression level of exosomal lncNORAD was detected by real-time quantitative polymerase chain reaction. In the BCa patient group, patients were divided into high expression and low expression groups based on the median relative expression level of lncNORAD. The χ2 test was used to compare the relationship between lncNORAD expression and clinical pathological characteristics, and the receiver operating characteristic (ROC) curve was used to evaluate its diagnostic efficacy for BCa.Results:The expression level of plasma exosomal lncNORAD in the patient group was significantly higher than that in the control group ( P<0.001) and the benign disease group ( P<0.001). High expression of lncNORAD was closely related to TNM stage ( P=0.001, χ2=10.187), lymph node metastasis ( P=0.003, χ2=8.575), tumor grade ( P=0.006, χ2=7.440), and nuclear matrix protein 22 (NMP22, χ2=5.844) ( P=0.016). ROC analysis showed that the area under the curve (AUC) for diagnosing BCa with plasma exosomal lncNORAD was 0.828 (95% CI 0.772-0.883). The diagnostic efficacy was further improved when exosomal lncNORAD was combined with the conventional marker NMP22 (AUC=0.866, 95% CI 0.815-0.916). Conclusion:The upregulation of lncNORAD in plasma exosomes of BCa patients is associated with multiple clinical pathological characteristics, indicating its potential as a diagnostic biomarker for BCa.

Background::Compared with human leukocyte antigen (HLA)-matched sibling donor (MSD) transplantation, it remains unclear whether haploidentical donor (HID) transplantation has a superior graft-versus-leukemia (GVL) effect for Philadelphia-negative (Ph-) high-risk B-cell acute lymphoblastic leukemia (B-ALL). This study aimed to compare the GVL effect between HID and MSD transplantation for Ph- high-risk B-ALL.Methods::This study population came from two prospective multicenter trials (NCT01883180, NCT02673008). Immunosuppressant withdrawal and prophylactic or pre-emptive donor lymphocyte infusion (DLI) were administered in patients without active graft-versus-host disease (GVHD) to prevent relapse. All patients with measurable residual disease (MRD) positivity posttransplantation (post-MRD+) or non-remission (NR) pre-transplantation received prophylactic/pre-emptive interventions. The primary endpoint was the incidence of post-MRD+.Results::A total of 335 patients with Ph- high-risk B-ALL were enrolled, including 145 and 190, respectively, in the HID and MSD groups. The 3-year cumulative incidence of post-MRD+ was 27.2% (95% confidence interval [CI]: 20.2%-34.7%) and 42.6% (35.5%-49.6%) in the HID and MSD groups (P = 0.003), respectively. A total of 156 patients received DLI, including 60 (41.4%) and 96 (50.5%), respectively, in the HID and MSD groups ( P= 0.096). The 3-year cumulative incidence of relapse was 18.6% (95% CI: 12.7%-25.4%) and 25.9% (19.9%-32.3%; P = 0.116) in the two groups, respectively. The 3-year overall survival (OS) was 67.4% (95% CI: 59.1%-74.4%) and 61.6% (54.2%-68.1%; P = 0.382), leukemia-free survival (LFS) was 63.4% (95% CI: 55.0%-70.7%) and 58.2% (50.8%-64.9%; P= 0.429), and GVHD-free/relapse-free survival (GRFS) was 51.7% (95% CI: 43.3%-59.5%) and 37.8% (30.9%-44.6%; P= 0.041), respectively, in the HID and MSD groups. Conclusion::HID transplantation has a lower incidence of post-MRD+ than MSD transplantation, suggesting that HID transplantation might have a superior GVL effect than MSD transplantation for Ph- high-risk B-ALL patients.Trial registration::ClinicalTrials.gov: NCT01883180, NCT02673008.

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.

To explore the expression of p62 protein in lung adenocarcinoma (LUAD). In this study, a cross-sectional study was adopted. From December 2011 to May 2013, 60 patients with lung adenocarcinoma who were diagnosed and treated in Tongji Hospital of Tongji University, Shanghai were selected for paraffin embedding and tissue chip preparation, and immunohistochemistry (IHC) technology was used to detect the expression of p62 in lung adenocarcinoma patients′ cancer tissues and adjacent tissues, and analyze the relationship between p62 expression and the clinicopathological characteristics and survival prognosis of patients with lung adenocarcinoma; at the same time, 6 cases of lung adenocarcinoma were selected by random sampling cancer tissues and adjacent tissues were detected by Western Blot (WB) to detect p62 protein and analyzed by gray value. Preoperative examination specimens of inpatients with lung adenocarcinoma diagnosed from April 2018 to early October 2019, and plasma specimens of healthy subjects were collected, and enzyme linked immunosorbent assay (ELISA) was used to detect lung adenocarcinoma patients and healthy patients. The expression of p62 in the plasma of the subjects was statistically analyzed using SPSS 22.0 software. The results of IHC showed that the positive expression rate of p62 in cancer tissues was significantly higher than that in adjacent tissues, and the difference was statistically significant ( t=5.593, P<0.001). Similarly, WB results showed that the expression of p62 protein in cancer tissues was significantly higher than that in adjacent tissues. It is statistically relevant ( t=2.238, P=0.049). The expression of p62 was statistically correlated with tumor size, clinicopathological stage and lymph node metastasis in patients with lung adenocarcinoma (all P<0.05). The overall survival of patients with lung adenocarcinoma with high p62 expression was worse than that of patients with low p62 expression (95 %CI was 0.238-0.870, P=0.028), suggesting that the high expression of p62 is related to the poor prognosis of patients with lung adenocarcinoma. The level of p62 protein in the plasma of patients with lung adenocarcinoma was significantly higher than that in the healthy control group. The difference was statistically significant ( t=8.533, P<0.001). The area under the receiver operating characteristic curve was 0.835 (95 %CI was 0.779-0.891, P<0.001), which is significantly higher than CEA, CA125, CA153 and other single traditional indicators, and the combined detection of four indicators has the highest diagnostic efficiency. p62 was strongly expressed in cancer tissues and serum, which is related to the poor prognosis and overall survival rate of LUAD patients.

To explore the expression of p62 protein in lung adenocarcinoma (LUAD). In this study, a cross-sectional study was adopted. From December 2011 to May 2013, 60 patients with lung adenocarcinoma who were diagnosed and treated in Tongji Hospital of Tongji University, Shanghai were selected for paraffin embedding and tissue chip preparation, and immunohistochemistry (IHC) technology was used to detect the expression of p62 in lung adenocarcinoma patients′ cancer tissues and adjacent tissues, and analyze the relationship between p62 expression and the clinicopathological characteristics and survival prognosis of patients with lung adenocarcinoma; at the same time, 6 cases of lung adenocarcinoma were selected by random sampling cancer tissues and adjacent tissues were detected by Western Blot (WB) to detect p62 protein and analyzed by gray value. Preoperative examination specimens of inpatients with lung adenocarcinoma diagnosed from April 2018 to early October 2019, and plasma specimens of healthy subjects were collected, and enzyme linked immunosorbent assay (ELISA) was used to detect lung adenocarcinoma patients and healthy patients. The expression of p62 in the plasma of the subjects was statistically analyzed using SPSS 22.0 software. The results of IHC showed that the positive expression rate of p62 in cancer tissues was significantly higher than that in adjacent tissues, and the difference was statistically significant ( t=5.593, P<0.001). Similarly, WB results showed that the expression of p62 protein in cancer tissues was significantly higher than that in adjacent tissues. It is statistically relevant ( t=2.238, P=0.049). The expression of p62 was statistically correlated with tumor size, clinicopathological stage and lymph node metastasis in patients with lung adenocarcinoma (all P<0.05). The overall survival of patients with lung adenocarcinoma with high p62 expression was worse than that of patients with low p62 expression (95 %CI was 0.238-0.870, P=0.028), suggesting that the high expression of p62 is related to the poor prognosis of patients with lung adenocarcinoma. The level of p62 protein in the plasma of patients with lung adenocarcinoma was significantly higher than that in the healthy control group. The difference was statistically significant ( t=8.533, P<0.001). The area under the receiver operating characteristic curve was 0.835 (95 %CI was 0.779-0.891, P<0.001), which is significantly higher than CEA, CA125, CA153 and other single traditional indicators, and the combined detection of four indicators has the highest diagnostic efficiency. p62 was strongly expressed in cancer tissues and serum, which is related to the poor prognosis and overall survival rate of LUAD patients.

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.

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.

Objective To observe the incidence of spinal cord injury (SCI) following aortic surgery in Fuwai Hospital of Chinese Academy of Medical Sciences, and evaluate the effect of standardized multidisciplinary spinal cord protection strategies, to summarize the experience in the prevention and treatment of SCI at perioperative period of aortic surgery. Methods The clinical data of patients underwent aortic surgery admitted to vascular center of Fuwai Hospital from January 2011 to December 2018 were retrospectively analyzed. The patients receiving traditional spinal cord protection strategies from January 2011 to December 2016 were defined as the control group, while the patients receiving standardized multidisciplinary spinal cord protection strategies from January 2017 to December 2018 were defined as the standardized treatment group. The standardized multidisciplinary treatment included preoperative cerebrospinal fluid drainage (CSFD), respiratory tract management, and maintenance of effective circulation of the lowest venous pressure; at the same time, anticoagulation, glucocorticoid, improve microcirculation, scavenge oxygen free radicals and other adjuvant treatments were started, and nerve function was monitored to prevent complications. The changes in SCI incidence after aortic surgery between the two groups were observed in order to evaluate the effect of standardized multidisciplinary spinal cord protection strategies. Meanwhile, the types of SCI after operation and the safety of CSFD were analyzed. Results During the 8-year period, 7 724 patients underwent aortic surgery at vascular center of Fuwai Hospital, 64 of which suffered from SCI after aortic surgery with total incidence of 0.83%. The onset of SCI was immediate in 39 patients (60.94%) and was delayed in 25 patients (39.06%), more than half of patients were immediate SCI. Of 64 patients with SCI, 52 patients (81.25%) underwent paraplegia and 12 (18.75%) underwent paraparesis. SCI persisted beyond discharge in 38 patients (59.38%) and 25 patients (39.06%) fully or partly recovered form SCI. One patient (1.56%) died. Compared with the control group, the incidence of SCI was decreased significantly after application of standardized multidisciplinary spinal cord protection strategies. The total incidence of SCI after aortic surgery was decreased from 1.06% (52/4 893) to 0.42% (12/2 831), the incidence after aortic arch replacement under deep hypothermic circulatory arrest was decreased from 3.66% (40/1 092) to 1.11% (5/450), and the incidence after thoraco-abdominal aortic replacement was decreased from 9.40% (11/117) to 2.47% (2/81) with statistically significant difference (all P < 0.05). Perioperative CSFD analysis showed that the incidence of CSFD-related complications was low, the overall incidence was 5.45% (3/55), including 1 patient of cerebrospinal fluid leakage, 2 patients of blood cerebrospinal fluid. No serious complications such as hemorrhage and infection of central nervous system occurred. Conclusions The main type of SCI after aortic surgery was immediate, about 39% SCI patients fully or partly recovered. Standardized multidisciplinary spinal cord protection strategies which included preoperative CSFD, reduced incidence of SCI after aortic surgery. The incidence of CSFD-related complications was low, which was safe and effective.

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.

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)