Objective:To investigate the effect of miR-22-3p and apoptosis-inducing factor,mitochondrion-associated 1(AIFM1)on mitochondrial homeostasis and apoptosis of pulmonary artery smooth muscle cells(PASMCs)under hypoxic conditions by establishing an in vitro model of pulmonary hypertension(PAH).Methods:PASMCs were cultured under hypoxic conditions to establish an in vitro model of PAH,and the expression levels of AIFM1 and miR-22-3p were upregulated or downregulated.Reverse transcription quantita-tive polymerase chain reaction(RT-qPCR)and Western blotting were used to measure the expression levels of AIFM1,miR-22-3p,and cleaved cysteine-aspartic protease-3(Cleaved Caspase-3).cell counting kit-8(CCK-8)assay was used to measure the prolifera-tive activity of cells,and flow cytometry was used to measure cell apoptosis.Mitochondrial superoxide indicator(mitoSOX)and adenos-ine triphosphate(ATP)assay kits were used to observe mitochondrial function and dynamics,and Mito-Tracker Red CMXRos(Mito-Tracker)was used to measure the change in mitochondrial circum-ference.Dual-luciferase reporter assay was used to validate the interaction between AIFM1 and miR-22-3p.Results:Hypoxia increased the content of mitochondrial ROS,reduced the level of ATP,promoted mitochondrial fission,and reduced cell apoptosis in PASMCs.AIFM1 overexpression improved mitochondrial homeo-stasis and increased cell apoptosis,while miR-22-3p negatively regulated AIFM1 and reversed the effect of AIFM1 overexpression.Conclusion:This study shows that miR-22-3p enhances mitochon-drial homeostasis,proliferation,and apoptosis in hypoxia-induced PASMCs by targeting AIFM1,which provides a potential theoretical basis for the prevention and treatment of PAH.

Objective:To analyze the clinical efficacy and prognostic factors of intracranial primary diffuse large B-cell lymphoma (DLBCL).Methods:Clinical data of 205 patients pathologically diagnosed with intracranial primary DLBCL at Sun Yat-sen University Cancer center from March 2001 to September 2020 were retrospectively analyzed. Among them, 101 patients were male and 104 female, the median age was 54 years old. Non-germinal center B cell (GCB) subtype accounted for 74.1%(126/170). A total of 177 patients received high-dose methotrexate (HD-MTX) and 91 patients received rituximab. After induction chemotherapy, 59 patients (30.4%) achieved complete response (CR), 112 patients (57.7%) achieved partial response (PR) or stable disease (SD). A total of 83 patients received consolidation or salvage radiotherapy, and only 14 patients received autologous stem cell transplantation (ASCT). The influence of pathological type, chemotherapy, rituximab treatment, radiotherapy and radiotherapy mode, ASCT and other factors on the overall survival (OS) and progression free survival (PFS) was evaluated. The survival rate was calculated by Kaplan-Meier method. Univariate prognostic analysis was performed by log-rank test. Multivariate prognostic analysis was conducted by COX model.Results:The median follow-up time was 34 months. The 5-year OS and PFS rates were 55.6% and 44.2%, respectively. GCB subtype, chemotherapy with HD-MTX, rituximab treatment, remission status after induction chemotherapy, and radiotherapy were favorable prognostic factors for OS or PFS, in which the last three were the independent prognostic factors. Consolidation radiotherapy in patients who obtained CR after induction chemotherapy did not significantly improve survival, while salvage radiotherapy in patients who achieved PR/SD after induction chemotherapy significantly improved both OS and PFS(both P<0.01). Consolidation radiotherapy showed no significant survival difference compared with consolidation ASCT. Conclusions:The non-GCB subtype of intracranial primary DLBCL is related to poor prognosis. The addition of rituximab to HD-MTX based induction chemotherapy can improve survival. Radiotherapy is still an important treatment for intracranial primary DLBCL, and there are limitations of ASCT in practical clinical application.

Objective:To explore and analyze the correlation between miR-31 in peripheral blood and oxidative stress indicators of diabetic nephropathy.Methods:A total of 94 patients with diabetic nephropathy who were admitted to Affiliated Hospital of Jining Medical College from September 2019 to September 2020 were selected. Patients were divided into mild diabetic nephropathy [estimated glomerular filtration rate (eGFR) 60-90 ml/min, 36 cases] group, moderate diabetic nephropathy (eGFR 30-60 ml/min, 27 cases) group and severe diabetic nephropathy (eGFR 0-30 ml/min, 31 cases) group according to the severity of the disease, and 30 healthy people in the same period were selected as the control group. Real time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-31 in peripheral blood. Serum superoxide dismutase (SOD), malondialdehyde (MDA), advanced oxidation protein products (AOPP) and other oxidative stress indicators, as well as serum urea nitrogen, creatinine and glomerular filtration rate. Pearson was used to analyze the correlation between peripheral blood miR-31 and oxidative stress indexes and renal function.Results:The expression of miR-31 in peripheral blood of patients with diabetic nephropathy was significantly lower than that in the control group, and the expression of miR-31 in peripheral blood of patients in severe and moderate diabetic nephropathy group was significantly lower than that in the mild diabetic nephropathy group (all P<0.05), with statistically significant difference (all P<0.05). Pearson correlation analysis showed that serum miR-31 expression was negatively correlated with the severity of diabetic nephropathy ( r=-0.526, P<0.05). The levels of serum MDA, SOD and AOPP in the diabetic nephropathy group were significantly higher than those in the control group, and the levels of serum MDA, SOD and AOPP in the severe and moderate diabetic nephropathy groups were higher than those in the mild diabetic nephropathy group, with statistically significant difference (all P<0.05). The levels of serum creatinine and blood urea nitrogen in the diabetic nephropathy group were higher than those in the control group, and the glomerular filtration rate was lower than that in the control group (all P<0.05). The levels of serum creatinine and blood urea nitrogen in the severe and moderate diabetic nephropathy group were higher than those in the mild diabetic nephropathy group, while the level of glomerular filtration rate was lower than that in the mild diabetic nephropathy group, with statistically significant difference (all P<0.05). Pearson correlation analysis showed that the expression of miR-31 in peripheral blood was negatively correlated with the levels of MDA, SOD, AOPP, serum creatinine and urea nitrogen (all P<0.05), but positively correlated with glomerular filtration rate ( P<0.05). Conclusions:The expression of miR-31 in peripheral blood gradually decreases with the severity of renal damage. Its level is negatively correlated with oxidative stress indicators of diabetic nephropathy, and positively correlated with glomerular filtration rate, which can be used for for clinical treatment and disease evaluation.

Objective:To investigate the effect of metformin on the microRNA (miRNA) expression and screen potential target with network pharmacology analysis in patients with type 2 diabetes.Methods:Fifteen patients with new diagnosed type 2 diabetes admitted to our hospital were selected, who received metformin during hospitalization and after discharge. The expression of serum matrix metalloproteinase (MMP)-9, transforming growth factor (TGF)-β1, and myocardial fibrosis related miRNAs were compared before and 6 month after metformin treatment. In addition, gene ontology (GO) and KEGG pathway enrichment analysis were applied to analyze differential expression miRNAs showing statistical significance. Meanwhile, the network figure was established to reflect the target gene messenger RNA (mRNA) corresponding to differentially expressed miRNA.Results:Compared with pre-medication, the serum level of MMP-9 was significantly decreased after treatment ( P<0.05). Besides, the expression of homo sapiens microRNA (hsa-miR)29a-3p, hsa-miR133a-5p, hsa-miR21-5p, hsa-miR30c-5p, and hsa-miR1-3p in patients with type 2 diabetes were dramatically down-regulated by metformin ( P<0.05 or P<0.01). Results of GO analysis and KEGG pathway enrichment analysis showed that differentially expressed miRNAs were mainly concentrated in endoplasmic reticulum lumen, synapse, basement membrane and other cell components. The molecular functions such as Rho GTPase binding and participation in extracellular matrix structural constituent were exerted through biological processes such as collagen catabolic process, regulation of short-term neuronal synaptic plasticity, and axon extension, which were mainly enriched in advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway in diabetic complications, tumor necrosis factor (TNF) signaling pathway, and Wnt signaling pathway, etc. The outcome of miRNA-mRNA network analysis demonstrated that there were 230 target genes mRNAs corresponding to differentially expressed miRNA. Conclusion:Metformin could play its role in the treatment of type 2 diabetes by down regulating the expression of miRNA, participating in the transduction of related cellular signaling pathways, regulating chromatin, nucleic acid binding, and enzyme activities.

Objective: To explore the blood lead level and its relationship with behavior in school-age children from rural areas of Chongqing. Methods: A total of 697 students from grades 3 to 6 in the fall semester of 2014 from 14 rural townships in one district of Chongqing was selected by using the random cluster sampling method. Blood were sampled to analyze the lead level. Neurobehavioral tests were performed to determine their personal cognitive and memory ability. Questionnaires and physical examinations were administered to obtain the information of confounding factors. All students were divided into Q₁-Q₄ groups according to the quartile of their blood lead level. The relationship between the blood lead level and behavior was analyzed by multivariate logistic regression model and restricted spline regression model. Results: The mean age of 697 students was (10.07±1.36) years old, and the median (interquartile range) of their blood lead level was 44.31 (35.42) μg/L. Multivariate logistic regression model showed that after adjusting for age, gender, body mass index and maternal culture level, compared with Q₁ group, the OR (95%CI) values of high digit symbol substitution test (DSST) scores and high overall memory quotient (MQ) scores in Q₃ group were 1.65 (1.01-2.70) and 2.10 (1.21-3.62), and the OR (95%CI) value of high long term memory (LTM) scores in Q₄ group was 0.53 (0.31-0.92). The results of the restricted spline regression model showed that the dose-response curves between the blood lead level and MQ/LTM test scores were both parabolic (P<0.05). Conclusion The blood lead level of school-age children from rural areas of Chongqing is the same as that from other areas of China, but slightly higher than that from other areas of Chongqing. Children with higher blood lead level have poor long-term memory ability.

Objective To explore the blood lead level and its relationship with behavior in school?age children from rural areas of Chongqing. Methods A total of 697 students from grades 3 to 6 in the fall semester of 2014 from 14 rural townships in one district of Chongqing was selected by using the random cluster sampling method. Blood were sampled to analyze the lead level. Neurobehavioral tests were performed to determine their personal cognitive and memory ability. Questionnaires and physical examinations were administered to obtain the information of confounding factors. All students were divided into Q1-Q4 groups according to the quartile of their blood lead level. The relationship between the blood lead level and behavior was analyzed by multivariate logistic regression model and restricted spline regression model. Results The mean age of 697 students was (10.07±1.36) years old, and the median (interquartile range) of their blood lead level was 44.31 (35.42) μg/L. Multivariate logistic regression model showed that after adjusting for age, gender, body mass index and maternal culture level, compared with Q1 group, the OR (95%CI ) values of high digit symbol substitution test (DSST) scores and high overall memory quotient (MQ) scores in Q 3 group were 1.65 (1.01-2.70) and 2.10 (1.21-3.62), and the OR (95%CI ) value of high long term memory (LTM) scores in Q4 group was 0.53 (0.31-0.92). The results of the restricted spline regression model showed that the dose?response curves between the blood lead level and MQ/LTM test scores were both parabolic (P<0.05). Conclusion The blood lead level of school?age children from rural areas of Chongqing is the same as that from other areas of China, but slightly higher than that from other areas of Chongqing. Children with higher blood lead level have poor long?term memory ability.

Objective To explore the blood lead level and its relationship with behavior in school?age children from rural areas of Chongqing. Methods A total of 697 students from grades 3 to 6 in the fall semester of 2014 from 14 rural townships in one district of Chongqing was selected by using the random cluster sampling method. Blood were sampled to analyze the lead level. Neurobehavioral tests were performed to determine their personal cognitive and memory ability. Questionnaires and physical examinations were administered to obtain the information of confounding factors. All students were divided into Q1-Q4 groups according to the quartile of their blood lead level. The relationship between the blood lead level and behavior was analyzed by multivariate logistic regression model and restricted spline regression model. Results The mean age of 697 students was (10.07±1.36) years old, and the median (interquartile range) of their blood lead level was 44.31 (35.42) μg/L. Multivariate logistic regression model showed that after adjusting for age, gender, body mass index and maternal culture level, compared with Q1 group, the OR (95%CI ) values of high digit symbol substitution test (DSST) scores and high overall memory quotient (MQ) scores in Q 3 group were 1.65 (1.01-2.70) and 2.10 (1.21-3.62), and the OR (95%CI ) value of high long term memory (LTM) scores in Q4 group was 0.53 (0.31-0.92). The results of the restricted spline regression model showed that the dose?response curves between the blood lead level and MQ/LTM test scores were both parabolic (P<0.05). Conclusion The blood lead level of school?age children from rural areas of Chongqing is the same as that from other areas of China, but slightly higher than that from other areas of Chongqing. Children with higher blood lead level have poor long?term memory ability.

Objective To estimate the diagnostic vaule of circulating tumor cells (CTCs) detection for epithelial ovarian cancer,and investigate the relationship between the presence of CTCs and the clinic pathologic characteristics of epithelial ovarian cancer patients.Methods Quantification of CTCs were performed using immunomagnetic bead based negative enrichment combined with flow cytometry in 65 patients with epithelial ovarian cancer,21 patients with benign ovarian diseases,and 10 healthy subjects.The differences among groups were analyzed by the Kruskal-Wallis H test (multi group comparison) and the Mann-Whitney U test (two group comparison),and the chi-square test was used in the positive rate comparison,the receiver operating characteristic (ROC) curve was established.The relationship between CTCs and clinic pathologic characteristics of epithelial ovarian cancer patients were analyzed.Results The receiver operating characteristic analysis showed the cut off value was 4.5 (＞ 4),the AUC was 0.806,and sensitivity,specificity and positive value (PPV) were 55.4 ％,96.8 ％ and 97.3 ％ respectively,in detecting patients with ovarian carcinoma malignancy.Quantification of CTCs in epithelial ovarian cancer was correlated with FIGO stages or distance metastasis (all P＜0.05),but not with patient age,histological types,pathologic differentiation,amount of ascites,tumor size and lymph node metastasis (all P＞0.05).Conclusion The detection of peripheral CTCs has a certain diagnosis value in epithelial ovarian cancer,especially with related to FIGO stages and distant metastasis.

Objective To establish a cell level-based negative enrichmen technique to detect circulating tumor cells (CTCs) in the peripheral blood of patients with epithelial ovarian cancer.Methods The colon cancer SKOV-3 cells were mixed with 2 ml whole blood from healthy donors at different ratio.Quantification of CTCs was performed using immunomagnetic bead based negative enrichment combined with immunofluorescence antibody method.The method was evaluated the recovery rate of target cells,Samples of 32 patients with ovarian cancer and 10 controls were assayed for CTCs detection by above method.Results ①The recovery rate was ranging from 64％ ～80％ by spiking varying numbers of SKOV-3 into 2 ml blood samples of healthy volunteers.Regression analysis of number of recovered SKOV-3 cells yielded a regression equation of Y=0.782X-1.408 and a coefficient of determination of R2 =0.998.②Did not detect CK8/18-+ circulating tumor cells in 10 controls,and CK8/18+ circulating tumor cells in 18 cases of 32 Patients with ovarian cancer.The positive rate of CK 8/18 + circulating tumor cells was significantly differences between the two groups (x2 =7.681,P＜0.01).③The presence ofCTCs was significantly correlated with distant metastasis (x2 =5.776,P＜0.05).Conclusion The method of immunomagnetic bead based negative enrichment combined with immunofluorescence antibody technique for CTCs detection in peripheral blood of patients with ovarian cancer has a clinical value of application and extension.

This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G(1) phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G(1) phase. L929 cells were sensitive to s-TNF-α when synchronized in G(1) phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G(1)/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; HL-60 Cells ; Hep G2 Cells ; Humans ; K562 Cells ; Membrane Proteins ; metabolism ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism ; pharmacology