1.Analyze the structure and component change of colonized bacterium in colorectal cancer tissues
Rong XU ; Zujun SUN ; Anquan SHANG ; Junlu WU ; Dong LI
Chinese Journal of Laboratory Medicine 2019;42(1):44-50
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.
2.Significance of combined detection of serum miR-21, interleukin-6 and interleukin-17 in the diagnosis of endometrial cancer
Juan HU ; Shenghaosong YAN ; Anquan SHANG
Chinese Journal of Primary Medicine and Pharmacy 2024;31(5):706-710
Objective:To study the significance and mechanism of miR-21, interleukin-6, interleukin-17 in the diagnosis of endometrial cancer.Methods:A retrospective analysis was condueted on the clinical data of 40 endometrial cancer patients and 40 healthy volunteers admitted to Lianyungang Second People's Hospital from March 2020 to March 2023. Real time fluorescence quantitative PCR was used to detect serum miR-21 levels and enzyme-linked immunosorbent assay was used to detect serum interleukin-6 and interleukin-17 levels in both groups.Results:The serum levels of interleukin-17 [(37.22 ± 13.08) ng/L], interleukin-6 [(12.09 ± 3.99) ng/L], and miR-21 [(2.83 ± 0.93)] in the endometrial cancer group were significantly higher than those in the normal control group [(15.72 ± 8.96) ng/L, (3.93 ± 2.19) ng/L, (1.20 ± 0.39), respectively] ( t = 8.58, 11.34, 10.20, all P < 0.001). Pearson correlation analysis showed that there was a significant positive correlation between serum interleukin-17 and interleukin-6 levels in patients with endometrial cancer ( r = 0.97, P < 0.001), between interleukin-17 level and miR-21 expression in patients with endometrial cancer ( r = 0.96, P < 0.001), and between interleukin-16 level and miR-21 expression in patients with endometrial cancer ( r = 0.99, P < 0.001). Receiver operating characteristic curve analysis showed that when the cut-off value of interleukin-17 was 21.65 ng/L, the sensitivity of the diagnosis of endometrial cancer was 80%, the specificity was 87.5%, and the area under the curve was 0.894 [95% confidence interval ( CI) = 0.827-0.961, P < 0.001]; when the cut-off value of interleukin-6 was 7.44 ng/L, the diagnostic sensitivity was 85.0%, the specificity was 92.5%, and the area under the curve was 0.959 (95% CI = 0.923-0.995, P < 0.001); when the cut-off value of miR-21 was 1.85, the diagnostic sensitivity was 77.5%, the specificity was 92.5%, and the area under the curve was 0.947 (95% CI = 0.905-0.989, P < 0.001). Conclusion:This study confirmed that serum interleukin-17, interleukin-6, and miR-21 could potentially serve as early predictors of endometrial cancer to a certain extent. The combined evaluation of these three parameters may be more valuable for diagnosis.
3.Expression level and clinical application value of human serum exosomal miR-218-5p in colorectal cancer
Wenjing CHANG ; Anquan SHANG ; Dianyu YANG ; Jiale TIAN ; Zujun SUN ; Dong LI ; Wenqiang QUAN
Chinese Journal of Laboratory Medicine 2021;44(8):709-714
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.
4.Prognostic value of serum carbamoyl phosphate synthase 1 in patients with hepatitis E-related acute liver failure
Jian WU ; Ze XIANG ; Guanghua ZHAI ; Anquan SHANG ; Wei LIANG ; Dong LI
Chinese Journal of Laboratory Medicine 2023;46(1):62-67
Objective:To explore the impact of serum carbamoyl phosphate synthase 1 (CPS1) level on prognosis of patients with hepatitis E-related acute liver failure (HEV-ALF).Methods:This retrospective analysis included 100 HEV-ALF patients, 100 patients with acute hepatitis E (AHE) and 100 healthy controls who admitted or underwent health checkup from December 2018 to June 2019 in six hospitals, including the First Affiliated Hospital, Zhejiang University School of Medicine. HEV-ALF patients were divided into non-survial ( n=21) and survival ( n=79) subgroups according to results of 30-day follow-up results. HEV-ALF patients were also divided into the high ( n=50) and low ( n=50) serum CPS1 level groups. HEV-ALF patients were further divided into the improvement ( n=55), fluctuation ( n=32) and deterioration ( n=13) subgroups. The general clinical data from all participants were collected. Serum CPS1 levels were detected by enzyme linked immunosorbent assay. The survival time in the high and low serum CPS1 level groups were presented in the Kaplan-Meier curve. The correlation between serum CPS1 level and HEV-ALF related conventional parameters was also analyzed by linear regression. The efficacy of serum CPS1 level on predicting the 30-day mortality of HEV-ALF patients was estimated by the receiver operating characteristic curve and area under curve (AUC). Results:Serum CPS1 level was significantly higher in HEV-ALF patients than in AHE patients [958.59 (665.52, 1 105.83) pg/ml vs 549.38 (495.02, 649.08) pg/ml, P<0.001], and serum CPS1 level was significantly higher in AHE patients than in healthy controls [549.38 (495.02, 649.08) pg/ml vs 469.89 (373.32, 564.53) pg/ml, P<0.001]. The level of serum CPS1 was significantly lower in the HEV-ALF survival group than in the HEV-ALF non-survival group [922.6 (652.7, 1, 042.3) pg/ml vs 1 252.8 (933.3, 1 555.8) pg/ml, P<0.001]. In addition, the survival time was shorter in the high serum CPS1 level group than in the low serum CPS1 level group [24.59 (22.11, 27.06) d vs 28.16 (26.25, 30.07) d, P=0.045]. Serum CPS1 levels were increased in the fluctuation and deterioration groups [Fluctuation: 1 328.3 (1 184.3, 1 964.0) pg/ml vs 1 245.7 (1 102.0, 1 937.6) pg/ml, P<0.01; Deterioration: 1 483.6 (1 275.9, 1 656.8) pg/ml vs 1 332.2 (1 197.4, 1 509.8) pg/ml, P<0.01], while decreased in the improvement group [810.3 (599.7, 904.5) pg/ml vs 922.6 (679.5, 1 039.6) pg/ml, P<0.01] over time. Besides, a linear positive correlation was found between serum CPS1 level and alanine aminotransferase (ALT) and total bilirubin (TBIL) (ALT: r=0.339, P<0.001; TBIL: r=0.304, P=0.002). The AUC of serum CPS1 level to predict the 30-day mortality of HEV-ALF patients was 0.803 (95% CI 0.666-0.941), the sensitivity and specificity were 66.67% and 97.47%, respectively. Conclusion:Serum CPS1 level was significantly increased in HEV-ALF patients, and closely related to the prognosis of patients with HEV-ALF.
5.Study on the expression of p62 protein in patients with lung adenocarcinoma
Bingjie ZENG ; Ping JI ; Zujun SUN ; Junlu WU ; Anquan SHANG ; Wenqiang QUAN ; Dong LI
Chinese Journal of Preventive Medicine 2021;55(5):685-690
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.
6.Study on the expression of p62 protein in patients with lung adenocarcinoma
Bingjie ZENG ; Ping JI ; Zujun SUN ; Junlu WU ; Anquan SHANG ; Wenqiang QUAN ; Dong LI
Chinese Journal of Preventive Medicine 2021;55(5):685-690
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.