1.Role and mechanism of circular RNA-vimentin in the proliferation and apoptosis of colorectal cancer cells
Jianning YAO ; Xuexiu ZHANG ; Yanzhen ZHANG ; Yanle LI ; Chunfeng WANG ; Lianfeng ZHANG
Chinese Journal of Digestion 2019;39(7):458-465
Objective To investigate the role and mechanism of circular RNA-vimentin (circ-VIM) in the proliferation and apoptosis of colorectal cancer cells.Methods From December 2016 to December 2017, at Department of General Surgery of The First Affiliated Hospital of Zhengzhou University, the clinical data of 100 patients who underwent radical resection of colorectal cancer and were confirmed by pathological examination after operation were collected.The tumor tissues and corresponding paracancerous tissues (negative control) were also collected.The expression of circ-VIM in the colorectal cancer tissues was determined by quantitative real-time polymerase chain reaction (qRT-PCR).The proliferation of HCT-116 and HT29 colorectal cancer cells was detected by cell counting kit-8 assay.The ratio of apoptosis of HCT-116 and HT29 cells was measured by annexin Ⅴ/propidium iodide double staining assay.The mitochondrial membrane potential of HCT-116 and HT29 cells was examined by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide (JC-1) assay.The expression changes of protein kinase B and mammalian target of rapamycin were tested by Western blotting.The target miRNA of circ-VIM was predicted by miRDB software.T-test and chi-square test were performed for statistical analysis. Results The expression of circ-VIM in colorectal cancer tissues was 2.387 ±0.536, which was higher than that in corresponding paracancerous tissues (1.110 ±0.134), and the difference was statistically significant (t =23.096, P <0.01).And the expression levels of circ-VIM were significantly different in patients with different tumor size, TNM stage and lymph node metastasis (all P <0.05).The proliferation of HCT-116 cells and HT29 cells in lenti-circ-VIM group was 0.737 ±0.023 and 0.835 ±0.025, respectively, which were both higher than those in control group (0.449 ±0.020 and 0.531 ±0.019), and the differences were statistically significant (t =20.706 and-15.374, both P <0.01).The proliferation of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was 0.236 ±0.027 and 0.243 ±0.019, which were lower than those in control group, and the differences were statistically significant (t =24.557 and -23.197, both P <0.01).The ratio of apoptosis of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was (18.00 ±1.82)% and (20.80 ±0.61)%, which was higher than those in control group ((6.64 ±2.01)% and (7.35 ±1.36)%), and the differences were statistically significant (t =8.826 and 17.454, both P <0.01).The fluorescence intensity ratio of JC-1 aggregate and JC-1 monomer of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was 2.21 ±0.12 and 1.40 ±0.11, which was lower than those in control group (14.54 ±1.00 and 9.24 ±1.18), and the differences were statistically significant (t =-19.558 and-15.685, both P <0.01), which indicated mitochondrial membrane potential decreased.After treated with lenti-circ-VIM-shRNA, the expression of phosphorylated protein kinase B, phosphorylated mammalian target of rapamycin, B-cell lymphoma-2 and mitochondrial cytochrome C at protein level were all down-regulated, however the expression of cytoplasmic cytochrome C, B-cell lymphoma-2 associated X protein and cleaved caspase-3 at protein level were all up-regulated.When the expression of circ-VIM was up-regulated, the expression of miRNA-147b, miRNA-4447 and miRNA-3656 was down-regulated.When the expression of circ-VIM was down-regulated, the expression of miRNA-147b, miRNA-4447 and miRNA-3656 was up-regulated.Conclusion The expression of circ-VIM in colorectal cancer is abnormally increased, which is involved in the proliferation and apoptosis of colorectal cancer cells.
2.Circular RNA-UBXN7 promotes proliferation, migration and suppresses apoptosis in hepatocellular cancer
Jianning YAO ; Xuexiu ZHANG ; Yanzhen ZHANG ; Yanle LI ; Chunfeng WANG ; Lianfeng ZHANG
Chinese Journal of Hepatology 2020;28(5):421-427
Objective:To investigate the effect and molecular mechanism of circular RNA-UBXN7 (circ_UBXN7) on the proliferation, migration and apoptosis of hepatocellular carcinoma cells.Methods:Circ_UBXN7 expression in the tissues and cells of hepatocellular cancer was detected by quantitative real-time polymerase chain reaction (qRT-PCR), and the relationship between circ_UBXN7 expression and clinicopathological features, including age, gender, tumor volume, pathological classification, staging, and lymph node metastasis was analyzed. The full-length sequence of circ_UBXN7 with lentivirus carrying lenti circ_UBXN7 and lenti circ_UBXN7 shRNA was constructed to transfect hepatocellular cell lines (HepG2 and Huh-7), respectively. CCK-8 experiments were performed to detect the ability of up- or down-regulation of circ_UBXN7 on the proliferation of HEPG2 and HUH-7 cells. Annexin V / PI experiment was used to detect the changes in apoptosis of HEPG2 and HUH-7 cells after up-regulation or down-regulation of circ_UBXN7 expression. JC-1 assay was used to detect the changes in mitochondrial potential energy of HEPG2 and HUH-7 cells after up-regulation or down-regulation of circ_UBXN7 expression. Transwell was used to detect the migration ability of HEPG2 and HUH-7 cells after up-regulation or down-regulation of circ_UBXN7 expression. Western blotting was used to detect the expressional change of TWIST, E-cadherin, N-cadherin and vimentin. Statistical analysis: The expression levels of circ_UBXN7 and clinicopathological features were measured by chi-square test. Two groups were compared by t-test and three groups and above were compared by single factor analysis of variance. LSD method was used for comparison between groups.Results:The expression of circ_UBXN7 in liver cancer tissues was significantly higher than adjacent tissues, and its expression level was significantly positively correlated with tumor volume, stage, and lymph node metastasis ( P < 0.05). Lenti-circ_UBXN7 had up-regulated the expression of circ_UBXN7 in HEPG2 and HUH-7 cells and promoted cell proliferation. Lenti-circ_UBXN7-shRNA had down-regulated the expression of circ_UBXN7 and induced apoptosis. Lenti-circ_UBXN7-shRNA had reduced the mitochondrial membrane potential of cells. Lenti-circ_UBXN7 had promoted cell migration, while lenti-circ_UBXN7-shRNA had inhibited cell migration. Lenti-circ_UBXN7 had induced increased expression of Twist, N-cadherin, and Vimentin proteins, and reduced the expression of E-cadherin protein. Lenti-circ_UBXN7-shRNA had opposite effects on the expression levels of each protein. Starbase V2.0 software showed that miR-203a and circ_UBXN7 had potential binding sites, and miR-203a and circ_UBXN7 expression levels were negatively correlated in HEP ??G2 and HUH-7 cells. Conclusion:circ_UBXN7 plays an important role in promoting the occurrence and development of liver cancer, and is expected to become a potential target for the treatment of liver cancer.
3. Clinicopathological Characteristics, Risk Factors of Perforation and Prognosis of Primary Small Intestinal Lymphoma: Analysis of 90 Cases
Fan FENG ; Xuexiu ZHANG ; Lianfeng ZHANG
Chinese Journal of Gastroenterology 2022;27(3):168-172
Background: Primary small intestinal lymphoma is an insidious onset gastrointestinal tumor with a high probability of perforation and poor prognosis. Aims: To investigate the clinicopathological characteristics and the factors related to perforation and prognosis in primary small intestinal lymphoma patients. Methods: The clinical data of patients with primary small intestinal lymphoma admitted to the First Affiliated Hospital of Zhengzhou University from January 2014 to January 2022 were collected retrospectively. The clinical features of patients with different pathological types were compared, and factors related to perforation and prognosis were identified by Logistic regression analysis and Cox regression analysis, respectively. Results: Ninety patients with primary small intestinal lymphoma were enrolled, the male to female ratio was 2∶1, and the median age was 52.5 years old. Abdominal pain was the most common symptom complained by patients (74.4%). All patients were diagnosed as non-Hodgkin’s lymphoma pathologically, of which 70 were B-cell lymphoma and 20 were T-cell lymphoma; diffuse large B-cell lymphoma was the most common histological type. Patients with T-cell lymphoma had a higher incidence of gastrointestinal complications than those with B-cell lymphoma (78.6% vs. 48.6%, P=0.001). Perforation was more commonly seen in T-cell lymphoma, and intestinal obstruction was more commonly seen in B-cell lymphoma. Multivariate Logistic analysis demonstrated that multisite involvement, elevation of lactate dehydrogenase (LDH), and T-cell lymphoma were the independent risk factors for perforation, while in univariate Cox regression analysis, decreased albumin, increased LDH, T-cell lymphoma, perforation and surgical treatment without chemotherapy were associated with poor prognosis. Furthermore, multivariate Cox regression analysis suggested that only surgical treatment without chemotherapy was an independent risk factor for death (HR=8.332, 95% CI: 1.453-47.772, P= 0.017). Conclusions: T - cell originated primary small intestinal lymphoma and those with increased LDH or involving multisite of gastrointestinal tract has a higher incidence of perforation. Surgical treatment without chemotherapy is strongly correlated with adverse outcomes. A regular chemotherapy after surgical treatment is highly recommended for primary small intestinal lymphoma patients complicated with perforation.
4.HCV infection and specific T lymphocyte immunity
Jijing SHI ; Jiyuan ZHANG ; Xuexiu ZHANG ; Fusheng WANG
Chinese Journal of Hepatology 2016;24(3):170-174
Adaptive immunity,especially the cellular immunity mediated by T lymphocytes,plays an important role in the body's fight against hepatitis C virus (HCV) infection.The clearance of HCV requires a strong,sustained,and multi-epitope response from specific T lymphocytes,and the dysfunction of specific T lymphocytes is the major reason for chronic infection.Effective treatment can restore the function of specific T lymphocytes to a certain extent.This article reviews the features and related mechanisms of immune response from specific T lymphocytes when the prognosis of patients with HCV infection is different.
5.Study on the risk factors of type 1 gastric neuroendocrine tumor in 123 cases of autoimmune gastritis
Mengge SU ; Xuyang DONG ; Xuexiu ZHANG ; Ya LI ; Haining ZHOU ; Xiangyu LI ; Jianning YAO
Chinese Journal of Digestion 2022;42(10):675-680
Objective:To investigate the risk factors of type 1 gastric neuroendocrine tumor (g-NET) in patients with autoimmune gastritis(AIG).Methods:From September 1, 2016 to February 28, 2022, 123 patients with AIG visited the First Affiliated Hospital of Zhengzhou University were retrospectively enrolled, including 37 cases with type 1 g-NET and 86 cases without type 1g-NET. The clinical data, serological indicators, and endoscopic manifestation of all the patients were analyzed, including the age at the time of AIG diagnosis (hereinafter referred to as the age at diagnosis), levels of gastrin 17 and pepsinogen Ⅰ (PGⅠ), presence or absence of gastric fundus and gastric body polyps, etc. The independent risk factors of type 1 g-NET in AIG patients were analyzed by univariate and multivariate logistic regression. The receiver operating characteristic curve (ROC) was plotted to analyze the optimal cut-off value, sensitivity and specificity of the independent risk factors in predicting type 1 g-NET in AIG patients. Independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:Compared with those of the AIG patients without type 1 g-NET, the age at diagnosis of AIG patients with type 1 g-NET was younger ((57.49±11.16) years old vs. (48.49±10.96) years old), the level of gastrin 17 was higher (200.21 ng/L, 121.85 ng/L to 244.40 ng/L vs. 244.40 ng/L, 182.50 ng/L to 248.02 ng/L), and the proportion of patients with gastric fundus and gastric body polyps was higher(18.6%, 16/86 vs. 56.8%, 21/37), and the differences were statistically significant( t=-4.13, Z=-3.06, χ2=17.90; P<0.001, =0.002 and <0.001). The results of univariate logistic analysis showed that the age at diagnosis ( OR=0.931, 95% confidence interval (95% CI)0.895 to 0.967), gastrin 17( OR=1.012, 95% CI 1.005 to 1.019), PGⅠ( OR=0.974, 95% CI 0.950 to 0.998)and gastric fundus and gastric body polyps( OR=5.742, 95% CI 2.461 to 13.399)were the influencing factors of type 1 g-NET in AIG patients ( P<0.001, =0.001, =0.033 and <0.001). The results of multivariate logistic regression analysis indicated that the age at diagnosis( OR=0.921, 95% CI 0.881 to 0.964), gastrin 17( OR=1.011, 95% CI 1.001 to 1.020), gastric fundus and gastric body polyps( OR=7.696, 95% CI 2.710 to 21.857)were the independent risk factors of type 1 g-NET in AIG patients ( P<0.001, =0.024 and <0.001). The results of ROC analysis demonstrated that the optimal cut-off values for the age at diagnosis and gastrin 17 in predicting type 1 g-NET were 56.50 years old and 206.40 ng/L, respectively; with sensitivity of 83.8% and 70.3%, respectively, and specificity of 54.7% for both ( P<0.001 and=0.003). Conclusion:The age at diagnosis< 56.50 years old, gastrin 17>206.40 ng/L and the presence of gastric fundus and gastric body polyps are independent risk factors of type 1 g-NET in AIG patients.
6.Clinical features and prognostic analysis of colorectal extranodal NK/T cell lymphoma
Xiangyu LI ; Jianning YAO ; Xuyang DONG ; Liangxing CHENG ; Xuexiu ZHANG ; Mengge SU ; Haining ZHOU ; Jinlin XIE ; Zhaoxiang SONG ; Fei HAN
Chinese Journal of Oncology 2024;46(8):782-793
Objective:To investigate the clinical manifestations, endoscopic characteristics, and prognostic factors of patients with colorectal extranodal NK/T cell lymphoma.Methods:The clinical data of 52 patients with colorectal extranodal NK/T cell lymphoma admitted to the First Affiliated Hospital of Zhengzhou University from January 2013 to January 2023 were retrospectively analyzed. Their clinical manifestations and endoscopic characteristics were summarized, and the prognostic factors were analyzed by Cox regression model.Results:Among the 52 patients with colorectal extranodal NK/T cell lymphoma, there were 35 males and 17 females, with a male-to-female ratio of 2.06∶1. Among the general symptoms, abdominal pain was the most common (39 cases), and B symptoms occurred in 47 patients, among which fever was the most common lymphoma B symptom (42 cases), and gastrointestinal perforation was the most common complication (18 cases). Forty-three patients underwent colonoscopy, and the main manifestations under endoscopy were the ulceration type (24 cases). The ulcers were irregular at the edges and often covered with moss at the bottom. The median survival time was 4.3 months. Multivariate Cox regression analysis showed that hemocytic syndrome ( HR=8.50,95% CI: 1.679-8.328, P=0.001), serum albumin ( HR=3.59,95% CI: 1.017-6.551, P=0.048), and with or without chemotherapy ( HR=0.31, 95% CI: 0.246-1.061, P=0.025) were independent factors influencing the overall survival of patients with colorectal extranodal NK/T cell lymphoma. Conclusions:Colorectal extranodal NK/T cell lymphoma is a rare disease with a very poor prognosis. When patients present with abdominal pain and lymphoma B symptoms, and when ulcers with irregular edges and moss covering the bottom are found under endoscopy, the disease should be considered, and endoscopic biopsy should be taken in time for pathological diagnosis. The prognosis of patients with hemophagocytic syndrome and hypoproteinemia is poor. This disease should be treated with chemotherapy and surgery, and on this basis, hemophagocytic syndrome and hypoproteinemia should be treated to improve the prognosis of patients.
7.Clinical features and prognostic analysis of colorectal extranodal NK/T cell lymphoma
Xiangyu LI ; Jianning YAO ; Xuyang DONG ; Liangxing CHENG ; Xuexiu ZHANG ; Mengge SU ; Haining ZHOU ; Jinlin XIE ; Zhaoxiang SONG ; Fei HAN
Chinese Journal of Oncology 2024;46(8):782-793
Objective:To investigate the clinical manifestations, endoscopic characteristics, and prognostic factors of patients with colorectal extranodal NK/T cell lymphoma.Methods:The clinical data of 52 patients with colorectal extranodal NK/T cell lymphoma admitted to the First Affiliated Hospital of Zhengzhou University from January 2013 to January 2023 were retrospectively analyzed. Their clinical manifestations and endoscopic characteristics were summarized, and the prognostic factors were analyzed by Cox regression model.Results:Among the 52 patients with colorectal extranodal NK/T cell lymphoma, there were 35 males and 17 females, with a male-to-female ratio of 2.06∶1. Among the general symptoms, abdominal pain was the most common (39 cases), and B symptoms occurred in 47 patients, among which fever was the most common lymphoma B symptom (42 cases), and gastrointestinal perforation was the most common complication (18 cases). Forty-three patients underwent colonoscopy, and the main manifestations under endoscopy were the ulceration type (24 cases). The ulcers were irregular at the edges and often covered with moss at the bottom. The median survival time was 4.3 months. Multivariate Cox regression analysis showed that hemocytic syndrome ( HR=8.50,95% CI: 1.679-8.328, P=0.001), serum albumin ( HR=3.59,95% CI: 1.017-6.551, P=0.048), and with or without chemotherapy ( HR=0.31, 95% CI: 0.246-1.061, P=0.025) were independent factors influencing the overall survival of patients with colorectal extranodal NK/T cell lymphoma. Conclusions:Colorectal extranodal NK/T cell lymphoma is a rare disease with a very poor prognosis. When patients present with abdominal pain and lymphoma B symptoms, and when ulcers with irregular edges and moss covering the bottom are found under endoscopy, the disease should be considered, and endoscopic biopsy should be taken in time for pathological diagnosis. The prognosis of patients with hemophagocytic syndrome and hypoproteinemia is poor. This disease should be treated with chemotherapy and surgery, and on this basis, hemophagocytic syndrome and hypoproteinemia should be treated to improve the prognosis of patients.
8.Adjunctive sepsis therapy with aminophylline (STAP): a randomized controlled trial
Ruifang ZHANG ; Huan LIU ; Dongmei DAI ; Xianfei DING ; Dong WANG ; Yan WANG ; Xuexiu SHI ; Shuguang ZHANG ; Xiaoguang DUAN ; Haixu WANG ; Yonggang LUO ; Shaohua LIU ; Bing HAN ; Xiaojuan ZHANG ; Yu FANG ; Jing YANG ; Wangbin XU ; Tongwen SUN
Chinese Medical Journal 2022;135(23):2843-2850
Background::Sepsis is a serious disease caused by infection. Aminophylline has anti-asthma and anti-inflammatory effects. We aimed to explore the safety and effect of aminophylline in sepsis.Methods::We conducted a clinical randomized controlled trial involving 100 patients diagnosed with sepsis within 48 h after intensive care unit (ICU) admission in two sites. All patients were randomized in a 1:1 ratio to receive standard therapy with or without aminophylline. The primary clinical outcome was all-cause mortality at 28 days.Results::From September 27, 2018 to February 12, 2020, we screened 277 septic patients and eventually enrolled 100 patients, with 50 assigned to the aminophylline group and 50 to the usual-care group. At 28 days, 7 of 50 patients (14.0%) in the aminophylline group had died, compared with 16 of 50 (32.0%) in the usual-care group ( P = 0.032). Cox regression showed that the aminophylline group had a lower hazard of death (hazard ratio = 0.312, 95% confidence interval: 0.129-0.753). Compared with the usual-care group, patients in the aminophylline group had a longer survival time ( P = 0.039 by the log-rank test). The effects of aminophylline on vasopressor dose, oxygenation index, and sequential organ failure assessment score were time-dependent with treatment. There were no significant differences in total hospitalization days, ICU hospitalization days, and rates of serious adverse events (all P > 0.05). No adverse events were observed in the trial. Conclusions::Aminophylline as an adjunct therapy could significantly reduce the risk of death and prolong the survival time of patients with sepsis.Trial registration::ChiCTR.org.cn, ChiCTR1800019173.