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@#Tumor immunotherapy is a therapeutic modality that uses immunological principles and methods to activate and enhance the body''s immune system to generate immune response for the removal of tumour cells. Many new immunotherapeutic agents have demonstrated effective anti-tumour capabilities, yet their clinical use is challenging due to the complex mechanisms of tumour immune escape. Meanwhile, these drugs would accumulate in different tissues and organs in the human body and be unable to achieve precise and specific targeting therapeutic effects, resulting in serious immune-related adverse effects, which greatly hinders the clinical potential of immunotherapy.Nanodrug delivery systems can deliver immunotherapeutic drugs to target tissues or specific immune cells precisely, thereby enhancing immune effects and reducing side effects.This paper reviews the research progress of nanodrug delivery systems in tumour immunotherapy in recent years based on the regulatory mechanism of the anti-tumour immune response, with a prospect of the challenges and development in this field.
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Objective:To investigate the value of SHOX2 and RASSF1A gene promoter region methylation detection for screening and diagnosis of early-stage lung adenocarcinoma.Methods:The mRNA sequencing data of 471 lung adenocarcinoma patients and corresponding methylation data of 413 cases were downloaded from The Cancer Genome Atlas (TCGA) database, the methylation levels of SHOX2 and RASSF1A gene promoter regions were calculated, and the difference in methy lation level between normal lung tissues and tumor tissues was analyzed. The clinical data of 54 patients with early-stage lung adenocarcinoma and 31 patients with benign lung tumors who underwent surgery at Drum Tower Hospital Affiliated to Nanjing University Medical School from January 2018 to January 2019 were retrospectively analyzed. The methylation status of SHOX2 and RASSF1A in tumor tissues and normal lung tissues (>5 cm from the edge of the tumor foci) (called clinical samples) was detect, and a positive methylation in the promoter region of either gene was considered as a combination of two genes methylation positivity. Using pathological diagnosis as the gold standard, the efficacy of gene methylation positivity in diagnosing early-stage lung adenocarcinoma was analyzed by receiver operating characteristic (ROC) curve. Patients with >80% of tumor cells in paraffin samples were screened, and mRNA high-throughput sequencing was performed in their tumor tissues and normal lung tissues. The relationship between positive methylation of the two genes and clinicopathological features was analyzed, and the correlation between the promoter region methylation level of the two genes and mRNA expression levels in clinical samples and TCGA database samples was analyzed by Spearman method. Gene set variance analysis (GSVA) method was used to analyze the differences in Kyoto Encyclopedia of Genes and Genomes enrichment pathways between two-gene methylation-positive clinical lung adenocarcinoma samples and corresponding methylation-negative lung adenocarcinoma.Results:In TCGA database, the SHOX2 promoter region methylation island contained 6 sequenced methylation sites, of which sites cg04532033 and cg01557547 methylation levels were higher in lung adenocarcinoma tissues than in normal lung tissues (both P < 0.05); the RASSF1A gene promoter region methylation island contained 11 sequenced methylation sites, and the methylation levels of 6 of these sites in lung adenocarcinoma tissues were higher than those in normal lung tissues (all P < 0.05). Compared with normal lung tissues, the methylation level of SHOX2 promoter region was higher in stage Ⅰ and Ⅱ lung adenocarcinoma tissues (both P < 0.05); the methylation level of RASSF1A promoter region was higher in all stages of lung adenocarcinoma ( P < 0.001). Among 54 patients with early-stage lung adenocarcinoma, 28 were positive for SHOX2 promoter region methylation in tumor tissues, 21 were positive for RASSF1A promoter region methylation, and 40 were positive for combined methylation of both genes; 31 benign lung nodules were negative for SHOX2 and RASSF1A methylation. ROC curve analysis showed that the sensitivity of positive SHOX2 promoter region methylation for diagnosing early-stage lung adenocarcinoma was higher than that of RASSF1A promoter region methylation positivity (51.8% vs. 38.9%), and the area under the curve (AUC) for diagnosis by two-gene methylation positivity was larger than that for diagnosis by SHOX2 or RASSF1A gene methylation positivity alone (0.870 vs. 0.759 and 0.694). The circulating thresholds (Ct) of SHOX2 and RASSF1A methylation tested by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) in stage Ⅰ and Ⅱ lung adenocarcinoma were lower than those in normal lung tissues (all P < 0.05); patients with two-gene methylation positivity were characterized by older age, longer tumor longest diameter and more advanced pathological stage compared with patients with two-gene methylation negativity (all P < 0.05). In clinical stage Ⅰ-Ⅱ lung adenocarcinoma samples, the Ct of SHOX2 and RASSF1A promoter region methylation tested by qRT-PCR was negatively correlated with their mRNA relative expression levels ( r=-0.43, P = 0.003; r = -0.48, P = 0.001); in TCGA database stage Ⅰ-Ⅱ lung adenocarcinoma samples, the level of SHOX2 promoter region methylation was negatively correlated with its mRNA relative expression level ( r = -0.23, P < 0.001), and the level of RASSF1A promoter region methylation was also negatively correlated with its mRNA relative expression level, but without statistical difference ( r = -0.05, P = 0.310). In two-gene promoter methylation-positive lung adenocarcinoma samples, the pathways related to folate metabolism and DNA stability were upregulated, and the pathways related to vasoconstriction and cell growth and differentiation were downregulated. Conclusions:The combined detection of SHOX2 and RASSF1A promoter region methylation can be used as an indicator for screening and diagnosis of early-stage lung adenocarcinoma. Abnormal promoter region methylation of the two genes may affect multiple tumor-related pathways and promote the occurrence and progression of early-stage lung adenocarcinoma.
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Objective:To compare the clinical characteristics and prognosis of bacterial liver abscess in patients with or without diabetes mellitus (DM), to provide a reference for clinical diagnosis and treatment.Methods:Patients with bacterial liver abscesses hospitalized in Tongren Hospital, Shanghai Jiao Tong University School of Medicine from January 2016 to August 2021 were enrolled, and their clinical data were collected. The patients were divided into diabetic and non-diabetic groups for comparison according to whether they had comorbid DM. Statistical analysis was performed by chi-square test or Fisher′s exact test, and multivariate logistic regression analysis.Results:A total of 131 patients with bacterial liver abscesses were included, including 47 cases in the diabetic group and 84 cases in the non-diabetic group. The percentages of platelet count <100×10 9/L, C-reactive protein>10 mg/L, and total bilirubin>17.5 μmol/L were lower in the diabetic group than that in the non-diabetic group, and the differences were all statistically significant ( χ2=3.90, 6.44 and 5.56, respectively, all P<0.05). The percentage of multiple abscesses in the diabetic group was 10.6%(5/47), which was lower than 29.8%(25/84) in the non-diabetic group, and the difference was statistically significant ( χ2=6.24, P=0.012). The positive rate of pus culture for Klebsiella pneumoniae was 64.9%(24/37) in the diabetic group, which was higher than 41.5%(27/65) in the non-diabetic group, with a statistically significant difference ( χ2=5.13, P=0.023). The incidences of pleural effusion and abscesses at other sites in the diabetic group were 29.8%(14/47) and 10.6%(5/47), respectively, which were both higher than 14.3%(12/84) and 1.2%(1/84) in the non-diabetic group, respectively, and the differences were statistically significant ( χ2=4.55, Fisher′s exact test, both P<0.05). The proportion of hospital stays>21 d was 34.0%(16/47) in the diabetic group, which was higher than 16.7%(14/84) in the non-diabetic group, with a statistically significant difference ( χ2=5.15, P=0.023). DM (odds ratio ( OR)=2.654, 95% confidence interval ( CI) 1.020 to 6.907, P=0.046) and abscess maximum diameter>10 cm ( OR=11.045, 95% CI 4.493 to 27.154, P<0.001) were significant risk factors for hospital stay>21 d. Conclusions:Bacterial liver abscesses combined with DM are more common with single abscess, a higher rate of Klebsiella pneumoniae infection, and more likely to develop pleural effusions and abscesses at other sites. Liver abscesses>10 cm in maximum diameter and comorbid DM would prolong hospital stay.
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Objective:To evaluate left ventricular systolic function and myocardial perfusion in patients with acute myocardial infarction (AMI) after percutaneous coronary intervention (PCI) by left ventricular pressure-strain loop (PSL).Methods:From August 2020 to December 2020, 47 patients with AMI admitted to the Intensive Care Unit of Cardiovascular Department of the First People′s Hospital of Foshan and treated with PCI were selected. Myocardial contrast echocardiography (MCE) and conventional echocardiography were performed within 72 hours after operation (T1 phase) and conventional echocardiography was repeated 3 months later (T2 phase). Myocardial perfusion scores (MPS) of left ventricular segments were obtained by MCE and the overall myocardial perfusion score index (PSI) was calculated. According to PSI, the patients were divided into good perfusion group and poor perfusion group. Conventional ultrasonic parameters and two-dimensional global longitudinal strain (2D-GLS) were collected. Left ventricular PSL analyzed in off-line EchoPAC software was used to evaluate the left ventricular myocardial work index, including global work index (GWI), global constructive work (GCW), global waste work (GWW), global work efficiency (GWE). The differences of parameters between patients with different perfusion levels and the change of parameters with time at the same perfusion level were compared. ROC curves were used to analyze the diagnostic values of strain parameters and myocardial work parameters in patients with hypoperfusion.Results:There were no significant differences in conventional ultrasound parameters between groups in T1 and T2 phases (all P>0.05), while there were significant differences in 2D-GLS and myocardial work parameters (except GWI in T2 phase) (all P<0.05). The absolute values of 2D-GLS and myocardial work parameters (except GWW ) were higher than those in T1 phase (all P<0.05). There was no significant difference in GWW ( P>0.05), but it decreased in good perfusion group while increased in poor perfusion group over time. ROC curve analysis showed that 2D-GLS, GWI, GCW and GWE had high area under the curve. Conclusions:Left ventricular PSL provides a new sensitive method for the evaluation of cardiac function in patients with AMI after PCI, and is expected to become a new index for the preliminary evaluation of microcirculation.
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Objective:To analyze the difference in immune microenvironment between primary tumor tissues and metastatic tumor tissues of metastatic colorectal cancer, and to screen specific immune-related differentially expressed genes (DEG) related to prognosis of metastatic colorectal cancer via bioinformatics methods.Methods:The GSE131418 microarray dataset of colorectal cancer and metastases was downloaded from gene expression omnibus (GEO) database, including 517 samples from the MCC cohort and 618 samples from the Consortium cohort in Moffitt Cancer Center. Immune-related gene sets were downloaded from immunology database and analysis portal IMMPORT, including 2 483 immune-related genes. A total of 695 cases of RNA sequencing data and 627 cases of clinical information of colorectal cancer tumors and adjacent tissues were downloaded from Cancer Genome Atlas (TCGA) data. The stroma cell score, immune cell score and stromal immune total score of metastatic tumor tissues and primary tumor tissues were calculated by using ESTIMATE algorithm, and 22 kinds of immune cell infiltration in primary tumor and metastatic tumor tissues of colorectal cancer were compared and analyzed by using CIBERSORT deconvolution algorithm. Immune-related DEG were screened to make Kyoto Encyclopedia of Genes and Gnomes (KEGG) signaling pathway enrichment analysis. The patients were divided into high and low expression groups according to the median expression levels of immune-related DEG. The Kaplan-Meier method and Cox regression risk model were used to analyze immune-related DEG, and the genes significantly related to prognosis in the results of the two methods were screened (all P < 0.01), and multivariate analysis was performed by using Cox regression method. The expression differences of each gene in tumor tissues, adjacent tissues, primary tumor tissues and metastatic tissues in GSE131418 data sets of TCGA database and GEO database were compared, and survival analysis was also performed. Results:The stroma cell score, immune cell score and stromal immune total score of colorectal cancer metastatic tissues were lower than those of primary tumor tissues (all P < 0.001). Compared with primary tumor tissues, the proportion of activated natural killer (NK) cells, monocytes, CD8 + T cells, T cells, activated dendritic cells in metastatic colorectal cancer tissues was increased, while the proportion of inactive mast cells, inactive dendritic cells, inactive NK cells, activated memory CD4 + T cells, M1 macrophages, and neutrophils was decreased. There were 289 immune-related DEG in metastatic tissues and primary tumor tissues of metastatic colorectal cancer, including 101 up-regulated genes and 188 down-regulated genes. KEGG signaling pathway enrichment analysis showed that in the immune microenvironment of metastatic tissues in metastatic colorectal cancer, vascular endothelial growth factor (VEGF) signaling pathway, programmed death ligand 1 (PD-L1) expression and programmed death 1 (PD-1) checkpoint pathway, T helper cell (Th) 1, Th2 and Th17 cell differentiation, NF-kappa B signaling pathway, interleukin 17 (IL-17) signaling pathway, chemokine signaling pathway, T cell receptor signaling pathway, MAPK signaling pathway, and NK cell-mediated cytotoxicity pathways enrichment were detected. Immune-related DEG related to prognosis including ANGPTL5, FPR1, HSPA8, NR2E3, PSMD2, PSMD8 and SBDS were screened out. Cox regression multivariate analysis showed that immune-related DEG ANGPTL5 ( HR = 2.69, 95% CI 1.22-5.92, P < 0.05), HSPA8 ( HR = 0.57, 95% CI 0.33-0.97, P < 0.05), and SBDS ( HR = 2.23, 95% CI 1.18-4.21, P < 0.05) were independent prognostic factors for metastatic colorectal cancer. The expression of ANGPTL5 in tumor tissues was lower than that in normal tissues, and the expression of ANGPTL5 in metastatic tissues was higher than that in primary tumor tissues. Patients with high expression of ANGPTL5 in tumor tissues had worse prognosis. The expression of HSPA8 in tumor tissues was higher than that in normal tissues, and the expression of HSPA8 in metastatic tissues was lower than that in primary tumor tissues. Patients with high expression of HSPA8 in tumor tissues had a better prognosis. The expression of SBDS in tumor tissues was lower than that in normal tissues, and the expression of SBDS in metastatic tissues was lower than that in primary tumor tissues. Patients with high expression of SBDS in tumor tissues had worse prognosis. Conclusions:Immune microenvironment of metastatic colorectal cancer is quite different from that of primary tumor. The degree of immune cell infiltration is reduced and the whole is immunosuppressed. The specific immune-related DEG related to prognosis of metastatic colorectal cancer may be new therapeutic targets of metastatic colorectal cancer.
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Dopaminergic neurons in the ventral tegmental area (VTA) play an important role in cognition, emergence from anesthesia, reward, and aversion, and their projection to the cortex is a crucial part of the "bottom-up" ascending activating system. The prelimbic cortex (PrL) is one of the important projection regions of the VTA. However, the roles of dopaminergic neurons in the VTA and the VTADA-PrL pathway under sevoflurane anesthesia in rats remain unclear. In this study, we found that intraperitoneal injection and local microinjection of a dopamine D1 receptor agonist (Chloro-APB) into the PrL had an emergence-promoting effect on sevoflurane anesthesia in rats, while injection of a dopamine D1 receptor antagonist (SCH23390) deepened anesthesia. The results of chemogenetics combined with microinjection and optogenetics showed that activating the VTADA-PrL pathway prolonged the induction time and shortened the emergence time of anesthesia. These results demonstrate that the dopaminergic system in the VTA has an emergence-promoting effect and that the bottom-up VTADA-PrL pathway facilitates emergence from sevoflurane anesthesia.
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Animals , Rats , Anesthesia , Dopaminergic Neurons/metabolism , Receptors, Dopamine D1/metabolism , Sevoflurane/pharmacology , Ventral Tegmental Area/metabolismABSTRACT
Objectvie:To investigate the infiltration patterns of immune cells in colorectal cancer, and to explore the correlation of immune cells infiltration with clinical characteristics and overall survival (OS) of patients.Methods:The RNA sequencing data of 615 patients with colorectal cancer were downloaded from The Cancer Genome Atlas (TCGA) database. The data was updated on July 19, 2019, and included 571 colorectal cancer tissues and 44 paracancerous tissues. There were 552 cases with clinical data, such as survival time, survival status, age, gender, clinical stage, grade, tumor location and so on. Using CIBERSORT deconvolution algorithm, the relative amounts of 22 immune cell types were calculated based on standardized gene expression data. According to the results of CIBERSORT algorithm, the samples with high accuracy of deconvolution result were selected ( P < 0.05), and they were used for analysis and graphing. The correlations between the infiltration patterns of immune cells and the clinical characteristics and OS of patients were analyzed. Results:After the CIBERSORT method was used to filter and remove samples with P ≥ 0.05, a total of 282 tumor tissue samples and 16 paracancerous tissue samples were screened. In 293 cases with clinical information, there were 277 tumor tissue samples and 16 paracancerous tissue samples. In 293 samples, M0 macrophages, M1 macrophages, M2 macrophages, CD8 + T cells and unactivated CD4 memory T cells accounted for a higher proportion of total immune cells; in tumor tissue samples, the expressions of M0 macrophages, M1 macrophages, activated CD4 memory T cells, and unactivated natural killer (NK) cells were higher; in paracancerous tissues, the expressions of naive B cells, M2 macrophages, activated NK cells, unactivated dendritic cells, unactivated mast cells and plasma cells were higher; with the increase of clinical stage, the expressions of follicular helper T cells, activated CD4 memory T cells, activated NK cells, M1 macrophages decreased, and the expressions of plasma cells and regulatory T cells increased, and the differences were statistically significant (all P < 0.05). M1 macrophages, unactivated mast cells, activated CD4 memory T cells, CD8 + T cells, and follicular helper T cells were highly expressed in right colon cancer, while M0 macrophages and activated mast cells were highly expressed in left colon and rectal cancer, and the differences were statistically significant (all P < 0.05). The patients were divided into high infiltration group and low infiltration group based on the median expression level of infiltrated immune cells, and the survival analysis was performed. The result of survival analysis showed that patients with high initial B cell infiltration had good OS; however, patients with high infiltration of M2 macrophages, activated mast cells, and neutrophils had poor OS. Conclusions:There are different types of immune cell infiltration patterns in the colorectal cancer samples of different stages and locations, which are closely related to tumor progression and OS of patients. They are expected to be applied to the development of therapeutic targets and prognosis prediction.
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Objective:To establish training system for postoperative delirium (POD) assessment and evaluate the efficacy of training for anesthesia nurses.Methods:Sixteen nurse anesthetists of both sexes in our hospital were selected and received the systemic training for POD assessment.The training system included questionnaire survey, theoretical teaching, simulated visit, clinical observation, independent evaluation, centralized question-answering, evaluation of efficacy and random inspection.The level of POD knowledge tests were performed before the training and at the end of the fourth week of independent evaluation, respectively.At week 1 and 4 of independent evaluation, the diagnostic rate of POD and sensitivity and specificity of the assessment were calculated, and Kappa consistency analysis was used to assess the consistency between anesthesia nurses and training group in diagnosis of POD.In the first week of the third month after the end of training, the evaluation results were randomly inspected, the POD diagnosis rate was calculated between the anesthesia nurses and the training group, and the consistency analysis was conducted.Results:Compared with the scores of POD knowledge questionnaire and sensitivity of the assessment of the anesthesia nurses in the first week of training, the scores were significantly increased ( P<0.05), and no significant change was found in the POD diagnosis rate in the fourth week of training ( P>0.05). Compared with the training group, the diagnosis rate of POD of anesthesia nurses was significantly decreased in the first week of training ( P<0.05), and no significant change was found at the fourth week of training ( P>0.05). In the first and fourth weeks of training, the Kappa value of anesthesia nurses and the training group was 0.676 and 0.954 ( P<0.001), respectively.In the first week of the third month after the end of training, the Kappa value between anesthesia nurses and the training group in diagnosis of POD was 0.862 ( P<0.05). Conclusion:The training system of POD assessment has been successfully established, and the standardized anesthesia nurses training of POD has been achieved with good results.
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Objective:To investigate the influence of infarct vessel diameter, blood flow before operation, blood flow status after operation, coronary artery disease, number of coronary artery lesions and other risk factors on the levels of inflammatory factors IL-1, IL-6, IL-10 in the coronary blood of patients with acute coronary syndrome (ACS), and to determine the relationship between inflammatory factors and ACS and its impact on clinical status.Methods:The peripheral blood and coronary blood of 54 patients with ACS underwent emergency interventional treatment were collected before the operation. The level of IL-1, IL-6, and IL-10 in the blood sample was detected by an automatic biochemical analyzer.Results:Postoperative adverse cardiac events were positively correlated with the patient's age, D-to-B time, and the number of coronary artery lesions (all P<0.05). The levels of IL-1, IL-6 and IL-10 in coronary blood of ACS patients were higher than those in peripheral blood (all P<0.05). The number of coronary artery disease branches (≥3), the onset time of myocardial infarction (>4 h), diabetes mellitus, ischemic post-treatment, preoperative blood flow level (<TIMI2 level) will increase the expression of IL-1 in serum (all P<0.05). For those who have no history of smoking, history of diabetes, infarct vessel diameter> 2.5 mm, coronary artery disease number ≥ 3, severe right coronary artery disease, preoperative blood flow <TIMI 2 level, postoperative CTFCs>22 frames, and cardiovascular events, the IL-10 levels are higher (all P<0.05). Conclusions:The levels of inflammatory factors IL-1, IL-6, and IL-10 in coronary blood of ACS patients are higher than those in peripheral blood, suggesting that acute coronary artery originates from local coronary inflammatory reaction, and the increase of inflammatory factors in peripheral blood belongs to "Erosion" effect. The level of inflammatory factors is significantly related to the diameter of the diseased blood vessel, preoperative blood flow, the number of coronary artery disease, the location of myocardial infarction, diabetes and smoking history.
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Cancer stem cells (CSCs) are a subpopulation of cancer cells with functions similar to those of normal stem cells. Although few in number, they are capable of self-renewal, unlimited proliferation, and multi-directional differentiation potential. In addition, CSCs have the ability to escape immune surveillance. Thus, they play an important role in the occurrence and development of tumors, and they are closely related to tumor invasion, metastasis, drug resistance, and recurrence after treatment. Therefore, specific targeting of CSCs may improve the efficiency of cancer therapy. A series of corresponding promising therapeutic strategies based on CSC targeting, such as the targeting of CSC niche, CSC signaling pathways, and CSC mitochondria, are currently under development. Given the rapid progression in this field and nanotechnology, drug delivery systems (DDSs) for CSC targeting are increasingly being developed. In this review, we summarize the advances in CSC-targeted DDSs. Furthermore, we highlight the latest developmental trends through the main line of CSC occurrence and development process; some considerations about the rationale, advantages, and limitations of different DDSs for CSC-targeted therapies were discussed.
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Objective:To investigate the application effect of kangaroo mother care on anxiety and parenting stress in premature mothers during hospitalization.Methods:From January to December 2018, 230 premature mothers were admitted to the neonatal intensive care unit of Ningxia Medical University General Hospital.They were randomly divided into the intervention group( n=114) and the control group( n=116). The premature infants in control group only implemented routine developmental support care.The premature infants in the intervention group was on the basis of routine developmental support care, 2 hours per day of kangaroo mother care was performed from the 2nd day incharge to discharge.Parental Stress Scale: Neonatal Intensive Care Unit and Parenting Stress Index-Short Form was performed using the 2nd day and the 14th day of hospitalization, and the two groups of premature mothers were evaluated for anxiety and parenting stress. Results:There were no significant differences in the scores of PSS: NICU((3.65±0.91) vs (3.63±0.91)) and PSI-SF((90.32±8.95) vs (90.12±7.17))(both P>0.05) between the two groups on the 2nd day of hospitalization (before intervention). On the 14th day of hospitalization (after intervention), the total scores ((2.57±0.80) vs (3.47±0.93))and each dimension scores of PSS: NICU in the intervention group were lower than those in the control group (first part (2.41±0.78) vs (3.81±0.73), second part (2.61±0.71) vs (3.14±0.83), third part (2.75±0.86) vs (3.57±1.06), fourth part(2.49±0.80) vs (3.35±0.94))( P<0.01). In addition, the total score of PSI-SF((79.09±8.02) vs (89.46±8.74)), parenting Distress((32.32±9.48) vs (37.15±9.67)), Parent-Child Dysfunctional Interaction((24.26±5.07) vs (27.34±5.26)), and Difficult Child scores((22.51±4.88) vs (24.97±5.05)) in the intervention group were lower than those in the control group( P<0.01). Conclusion:Kangaroo mother care can reduce the anxiety of premature mothers during hospitalization and help relieve the parenting stress of premature mothers.
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Background: Long non-coding RNA (lncRNA) could participate in the process of tumorigenesis by regulating the expression of miRNA, but the molecular mechanism of lncRNA in the development and metastasis of colorectal cancer has not been elucidated. Aims: To investigate the molecular mechanism of lncRNA SOX21-AS1 affecting the biological process of colorectal cancer cells by regulating the expression of microRNA-202-5p (miR-202-5p). Methods: The colorectal cancer HCT116 and SW480 cells were transfected with SOX21-AS1 small interfering RNA (si-SOX21-AS1), miR-202-5p mimics and its inhibitor. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expressions of SOX21-AS1 and miR-202-5p. Cell proliferation was detected by MTT assay. Flow cytometry was used to detect cell apoptosis. Transwell experiments were used to detect cell migration and invasion. Dual luciferase reporter assay was used to validate the targeted regulatory relationship between SOX21-AS1 and miR-202-5p. Western blotting was used to detect the protein expressions of cyclin D1, MMP-2 and cleaved caspase-3. Results: The mRNA expression of SOX21-AS1 in colorectal cancer cells was significantly higher than that in normal colonic mucosal epithelial cells (P<0.05), while the mRNA expression of miR-202-5p was significantly decreased (P<0.05). Silencing SOX21-AS1 or up-regulating miR-202-5p significantly inhibited proliferation, migration and invasion of HCT116, SW480 cells, the cell apoptosis rate was significantly increased (P<0.05), and significantly inhibited the protein expressions of cyclin D1 and MMP-2 (P<0.05), however, the protein expression of cleaved caspase-3 was significantly increased (P<0.05). SOX21-AS1 could bind to miR-202-5p and negatively regulate the expression and activity of miR-202-5p. Inhibition of miR-202-5p expression reversed the effect of silencing SOX21-AS1 on proliferation, apoptosis, migration and invasion of colorectal cancer cells. Conclusions: Silencing SOX21-AS1 inhibits the proliferation, migration and invasion of colorectal cancer cells and induces cell apoptosis by up-regulating the expression of miR-202-5p.
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Objective To evaluate the diagnostic value of CEA, CA19-9, CA72-4, AFP, and CA125 in gastric stromal tumors. Methods 41 patients with gastric stromal tumors and 11 patients with gastric leiomyoma were recruited in this study from Qilu Hospital of Shandong University during May 2014 to December 2017. The tissue was collected by surgery, and HE staining was done. Immunohistochemistry was employed to detect the expression of CD34, CD117, and DOG-1. Serum of all cases and 41 healthy volunteers in the same hospital were collected. The levels of CEA, CA19-9, CA72-4, AFP and CA125 were examined by electrochemiluminescence assay, and the differences in each group were compared by M-W test or K-W test. Then the relationship between those biomarkers and the clinical parameters of gastric stromal tumors was analyzed. Moreover, AUC (Area Under the curve), cut-off value, sensitivity, specificity, positive predictive value, negative predictive value with drawing ROC curve (receiver operating characteristic curve) were also calculated. Results Spindle cells or epithelioid cells were observed in the tissue of gastric stromal tumors. The expression of CD34, CD117, DOG-1 were positive. The level of the serum CEA 1.53 (1.15, 2.22) ng/ml in patients with gastric stromal tumor patients was higher than that in healthy controls 1.06 (0.62, 1.48) ng/ml and that in patients with gastric leiomyoma 0.79 (0.39, 1.39) ng/ml (the U value was 446.5, and 113.0 respectively, P<0.05). The level of CA19-9 in gastric stromal tumors 9.30 (4.95, 12.70) U/ml was higher than that in healthy controls 6.62 (4.56, 8.82) U/ml (the U value was 615.5, P<0.05). There were no significant differences in AFP, CA125, and CA72-4 of three groups (the H value was 4.348, 1.073, and 3.897, P>0.05). Furthermore, the level of CEA was closely related to TNM stage (the U value was 129.0, P<0.05). The diagnostic value of CEA and CA19-9 was statistically not significant (P>0.05). However, the combination of two markers might increase diagnostic efficiency. The sensitivity, specificity, positive predictive value, negative predictive value and AUC was 92.7％, 48.8％, 64.4％, 87.0％ and 0.752 respectively. Conclusion The combination of CEA and CA19-9 has better sensitivity and negative predictive value in auxiliary diagnosis of gastric stromal tumors.
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Tumor-associated fibroblasts(TAFs),the most important stromal cells of the tumor microenvironment (TME),have been found to support tumorigenesis and tumor metastasis in a variety of ways,including paracrine, direct contact with cells,immune regulation and extracellular matrix remolding.Therefore,TAFs in the TME have been an optimal target for cancer therapy.In this review,the TAFs targeted therapies are summarized to provide the new strategy for tumor treatments based on the analysis of the location and specific biological phenotypes of TAFs in tumors.
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This report summarized current knowledge and findings relevant to environmental and genetic risk factors in brain tumors,with a particular focus on glioma.To date,the established risk factors for brain tumors are family history and ionizing radiation exposure;whereas there is an inverse association between tumors and other factors such as history of allergies,atopic conditions,chickenpox,and varicella zoster virus infection.To identify inherited genetic variants impacting susceptibility of brain tumors,large scale familial linkage-scan pedigree analysis,populationbased candidate genes,and genome-wide association study were performed.More recently;next generation exome and whole genome sequencing studies were also conducted.
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CRM197 (cross-reacting material 197), a non-toxic mutant of diphtheria toxin, has wide application potential in biopharmaceuticals. However, it is difficult to express CRM197 in bacteria other than Corynebacterium diphtheriae. Here we proposed a new alternative method to produce soluble CRM197 without label in Escherichia coli. In particular, a synthetic gene coding for CRM197, optimized for E. coli codon usage, was cloned in the pET32a (+) vector. Accordingly, the over-expression of the protein was simply induced with IPTG in E. coli BL21 (DE3). The target protein was soluble and accounted for about 40% of the total protein in the supernatant. Following an ultrasonic cytolysis step, the recombinant protein was purified by anion exchange, affinity and desalting chromatography and the purity of the final preparation reached 95%. Cytotoxicity tests showed that the IC₅₀ value of CRM197 was 2.1×10⁷ times the IC₅₀ value of diphtheria toxin, and 9.6 times the IC50 value of diphtheria toxoid, telling that the target protein is safe and non-toxic. Subsequently, we found that both the high dose (20 μg) and the low dose (2 μg) of CRM197 were equally efficient in inducing an immune response against diphtheria toxiod in mice, and the antibodies titer of mice after three immunizations with low dose could reach 1:409 600. In conclusion, our findings provide a highly efficient strategy for the rapid production and purification of unlabeled and soluble recombinant CRM197 in E. coli, with good immunogenicity and safety.
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Objective To study in the correlation of the laboratory markers of coagulation,fibrinolysis and thrombosis in patients with sepsis and SOFA score,the procalcitonin (PCT) concentration and seven-day survival rate.Methods From February 2017 to March 2018,119 patients with sepsis admitted in ICU and another 119 patients with non-sepsis undergoing selective surgery were enrolled as control in this study.APTT (activated partial thromboplastin time),PT-INR (prothrombin time-international normalized ratio),FIB (fibrinogen),AT-Ⅲ (antithrombin Ⅲ),D-Dimer,FDP (fibrinogen degradation products),sTM (soluble thrombomodulin),TAT (thrombin antithrombin complex),PIC (plasmin-a2 plasminogen inhibitor complex) and t-PAI-C (tissue plasminogen activator and its inhibitor complex),were simultaneously monitored at admission.The correlation between the given laboratory markers mentioned and SOFA score,the PCT concentration and seven-day survival rate were analyzed with the Spearman correlation analysis.Results (① In the patients with sepsis,a positive correlation between SOFA score and sTM,t-PAI-C,TAT respectively was found,and a negative correlation between SOFA score and PLT (platelet count) was observed,and no correlation between SOFA score and PIC was noticed.(②) A positive correlation between PCT and sTM,t-PAI-C respectively was significant,a negative correlation between PCT and PLT was marked,and no correlation between PCT and AT-Ⅲ,TAT,PIC respectively was found.(③) A negative correlation between seven-day survival rate and sTM,t-PAI-C and TAT respectively was obvious,a positive correlation between seven-day survival rate and AT-Ⅲ,PLT respectively was occurred,and no correlation between seven-day survival rate and PIC was determined.Conclusions Soluble thrombomodulin (sTM),thrombin-antithrombin (TAT),antithrombin Ⅲ (AT-Ⅲ) and tissue plasminogen activator inhibitor complex (t-PAI-c) were good clinical monitoring indicators of coagulation disorder in patients with sepsis,which were the representative of the endothelial cell damage with highly activated coagulation,relatively insufficient anti-coagulation function and poor fibrin degradation ability.These were good adjuvants to PLT,INR and APTT for core diagnostic criteria of coagulation disorder in sepsis.
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Objective To investigate the diagnostic value of Enzymes linked immunosorbent assay(ELISA)detection of anti phospholipase A2 receptor antibody(anti-PLA2R)in idiopathic membranous nephropathy patients.Methods All articles published from January 2012 to April 2017 about ELISA test on anti phospholipase A2 recepeor antibody as a maker for IMN were collected, and the quality of the studies was evaluated by using the guidelines of the updated Quality Assessment of Diagnostic Accuracy Stud-ies(QUADAS-2)tool.According to the inclusion criteria and exclusion criteria,the research objects were selected and the data were extracted,and the analysis was carried out by Meta-Disc1.4 software.The results are summarized on the basis of sensitivity,speci-ficity,and summary receiver operating characteristic curve(ROC curve).Results Ten articles were selected according to inclusion criteria and 1517 cases were included in the study.The combined sensitivity was 0.68[95% CI(0.64,0.70)];the merge specificity was 0.92[95% CI(0.90,0.95)],and the subjects'working curve showed AUC 0.807 4.Conclusion The detection of anti-PLA2R by ELISA is effective in the diagnosis of idiopathic membranous nephropathy,which can be used as an effective supplement for the diagnosis of idiopathic membranous nephropathy.
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Objective To establish the pipeline and evaluate the feasibility of high-throughput sequencing method used in the detection of severe pneumonia pathogens.Methods Clinical control study was used.Bronchi alveolar lavage fluids (BALF) samples from 76 patients with severe pneumonia (treatment group) and 18 patients with tracheal malacia (control group) and without clinical detected pathogens were collected during March 2015 to December 2016 in Shenzhen Children′s Hospital.The pathogens in the samples were detected using clinical tests and high-throughput sequencing respectively.The results of high-throughput sequencing were confirmed by real-time quantitative PCR and the high-throughput sequencing method used in the detection of severe pneumonia pathogens was evaluated.The χ2 test was used to analyze the correlation of detection rate between the high-throughput sequencing group and the non high-throughput sequencing group.Results The pipeline and method of high-throughput sequencing used in the severe pneumonia pathogens detection was established.The pipeline included sample collection, DNA extraction, library construction, sequencing, and bioinformatic analysis.In 76 cases of patients with severe pneumonia, the results of high-throughput sequencing in 66 cases of bronchoalveolar lavage fluid specimens were positive.The sensitivity was 86.84%, which was significantly higher than the total sensitivity of traditional clinical detection methods including bacterial culture, immunofluorescence and quantitative PCR(68.42%,52/76),χ2=7.426,P<0.001.A total of 13 pathogens were detected in 66 positive samples of high-throughput sequencing, including Mycoplasma pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae and adenovirus, etc.Nine kinds of pathogens were detected in these samples through non-high-throughput sequencing.In the experimental group, the results obtained by clinical test and high-throughput (80.26%) were entirely consistent in 61 samples and not completely consistent in 15 samples (19.74%) specimens.These inconsistent results were mainly concentrated in the detection of adenovirus, Streptococcus pneumoniae and Haemophilus influenzae through high-throughput sequencing, whereas clinical cultures and immunofluorescence methods were not able to detect these pathogens.PCR validation showed that there was no significant difference between the results of high-throughput sequencing and the PCR tests (χ2=0.517,P=0.472), and the difference between the results of high-throughput sequencing and traditional clinical detection methods was statistically significant (χ2=11.671,P<0.001).Conclusion The method for the detection of severe pneumonia pathogens based on high-throughput sequencing is highly sensitive and can detect unknown pathogens, which is superior to those used in traditional clinical detection.
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Objective To explore the influence of smoking on clopidogrel resistance in patients with coro-nary artery disease. Methods A total of 216 patients with coronary artery disease who accept selective percutane-ous coronary intervention in our hospital from May 2015 to December 2015 were selected as study subjects;their average age was 63 years old ,146 were male and 70 were female. All the patients were divided into clopidogrel resistance group (CR group) and normal clopidogrel response group (NCR group) according to the results of platelet aggregation test. History of smoking ,alcohol drinking and diabetes mellitus ,baseline medication uses , level of platelet counts,platelet distribution width,mean platelet volume,HbA1c,and results of echocardiogram and coronary angiography were compared between the two groups. Results The rate of smoking was significantly higher in CR group than in NCR group (P < 0.05). The changed value of PDW before and after treatment with clopidogrel was smaller in CR group than in NCR group ,the difference was statistically significant (P < 0.05). Conclusions Smoking may be a protective factor for clopidogrel resistance. The changed value of PDW before and after treatment with clopidogrel can reflect the level of clopidogrel resistance.