MicroRNA 98-5p Overexpression Contributes to Delayed Fracture Healing via Targeting BMP-2.

MicroRNAs (miRNAs) are related to the regulation of bone metabolism. Delayed fracture healing (DFH) is a common complication after fracture surgery. The study attempted to examine the role of miR-98-5p and bone morphogenetic protein (BMP)-2 with the onset of DFH. A total of 140 patients with femoral neck fracture were recruited, including 80 cases with normal fracture healing (NFH) and 60 cases with DFH. MC3T3-E1 cells were induced cell differentiation for cell function experiments. Real-time quantitative polymerase chain reaction (RT-qPCR) was carried out to test mRNA levels. Cell proliferation and apoptosis were determined via CCK-8 and flow cytometry assay. Luciferase reporter assay was done to verify the targeted regulatory relationship of miR-98-5p with BMP-2. In comparison with NFH cases, DFH patients owned high levels of serum miR-98-5p and low concentration of BMP-2, and the levels of the two indexes are significantly negatively correlated. Both miR-98-5p and BMP-2 had the ability to predict DFH, while their combined diagnostic value is the highest. BMP-2 was demonstrated to be the target gene of miR-98-5p. Overexpression of BMP-2 reversed the role of miR-98-5p in MC3T3-E1 cell proliferation, apoptosis and differentiation. Increased miR-98-5p and decreased BMP-2 serve as potential biomarkers for the diagnosis of DFH. MiR-98-5p overexpression inhibits osteoblast proliferation and differentiation via targeting BMP-2.

Down-regulation of CX43 expression by miR-1 inhibits the proliferation and invasion of glioma cells.

Background: Connexin (CX) 43 makes glioblastoma resistant to temozolomide, the first-line chemotherapy drug. However, targeting CX43 is very difficult because the mechanisms underlying CX43-mediated resistance remain unclear. CX43 is highly expressed in glioblastoma, which is closely associated with poor prognosis and chemotherapy resistance. The present study was to analyze the mechanism of microRNA (miR)-1 in regulating the proliferation and invasion of glioma cells. Methods: The effects of knockdown of miR-1 on the growth of glioma cell lines were observed by establishing blank, miR-1 inhibitor, and miR-1 mimic groups. Cell proliferation was detected using a Cell Counting Kit-8 (CCK-8) assay, cell apoptosis was detected by flow cytometry, and protein expression was detected by western blot. We used the Student's t-test to assess continuous data between the two groups and the Kruskal-Wallis test was adopted for multiple group comparisons. Results: Compared with the mimics normal control (NC) group, the apoptosis rate of the miR-1-3p mimics group was decreased, while that of the miR-1-3p inhibitor group was increased compared to the inhibitor NC group. In addition, the miR-1-3p mimics model of U251 cells exerted an inhibitory effect on the invasion ability of cells, whereas the miR-1-3p inhibitor model of U251 cells showed an invasion-promoting effect. The dual-luciferase assay showed that miR-1-3p had a targeted relationship with the CX43 gene. Conclusions: Down-regulation of CX43 expression by miR-1 inhibited the infiltration and growth of glioma cells and further promoted the apoptosis of glioma cells by regulating CX43 expression.

[Expression of miR-126 in Diffuse Large B-Cell Lymphoma and Its Biological Function].

OBJECTIVE: To investigate the expression of miR-126 in diffuse large B-cell lymphoma (DLBCL) tissues and its biological function. METHODS: The lymphoma tissues of 46 DLBCL patients in our hospital were selected as the research object, and the lymph node hyperplasia tissue of 31 patients with reactive hyperplasia were selected as controls. The expression level of miR-126 in the patients' tissues was detected by real-time fluorescent quantitative PCR (RT-qPCR), and the correlation of miR-126 expression with the pathological characteristics and prognosis of the patients was analyzed. The DLBCL cell line SU-DHL-4 was transfected with miR-126 inhibitor and its negative control (NC inhibitor) or miR-126 mimics and its negative control (NC mimics). RT-qPCR assay was used to detect the expression level of miR-126 in cells; MTT method was used to detect cell proliferation activity; single clone formation test was used to detect cells colony-forming ability; Annexin V/PI double staining assay was used to detect cell apoptosis; Transwell test was used to detect cell migration and invasion ability; the expression levels of apoptosis-related proteins cleaved-Caspase-3, Bcl-2 and Bax were detected by Western blot. RESULTS: miR-126 was highly expressed in lymphoma tissues of DLBCL patients, and its expression level was significantly correlated with Hans type, IPI score and Ann-Arbor stage of DLBCL patients (P<0.05). Kaplan-Meier survival analysis showed that the survival rate of DLBCL patients with high expression of miR-126 was significantly lower than that of patients with low expression (P<0.05). Compared with the NC mimics group, the miR-126 expression level, cell proliferation rate, number of colony-forming units, migration and invasion ability, and Bcl-2 protein expression level in the miR-126 mimics group were significantly increased (P<0.05), but the cells apoptotic rate, cleaved-Caspase-3 and Bax protein expression levels were significantly reduced (P<0.05). Compared with the NC inhibitor group, the miR-126 expression level, cell proliferation rate, number of colony-forming units, migration and invasion ability, and Bcl-2 protein expression level in the miR-126 inhibitor group were significantly reduced (P<0.05), but the cells apoptosis rate, cleaved-Caspase-3 and Bax protein expression levels were significantly increased (P<0.05). CONCLUSION: miR-126 is highly expressed in lymphoma tissues of DLBCL patients and its expression level is related to the poor prognosis of patients. miR-126 can promote DLBCL cell proliferation, invasion and migration, and inhibit cell apoptosis.

[Risk Factors Analysis of Thromboembolism in Patients with Lymphoma Chemotherapy].

OBJECTIVE: To evaluate the risk factors affecting thromboembolism in lymphoma patients with chemotherapy. METHODS: Three hundred and four consecutive lymphoma patients treated by chemotherapy between January 2012 and July 2019 were enrolled and retrospectively analyzed, consisting of 111 patients with thromboembolism and 193 without thromboembolism. Univariate analysis was used to compare the clinical characteristics and related laboratory examination between the patients, while multivariate Logistic regression analysis were used to identify the risk factors affecting thromboembolism in lymphoma patients with chemotherapy. RESULTS: Univariate analysis showed that the female, BMI <18.5 or >24, ≥60 years old, with abnormal platelets before chemotherapy, prolonged single hospitalization days and patients at Ann Arbor stage III and IV could increase the incidence of thromboembolism in lymphoma patients treated by chemotherapy. Multivariate Logistic regression analysis showed that abnormal platelet count before chemotherapy, patients at Ann Arbor stage III and IV, and female were all the independent risk factors affecting thromboembolism in lymphoma patients thromboembolism after chemotherapy (P<0.05). CONCLUSION: For lymphoma chemotherapy patients, female, abnormal platelet count before chemotherapy and Ann Arbor stages III and IV show a significantly higher risk for thromboembolism. Thus, preventive anticoagulation therapy is recommended.

Risk Factors of Thromboembolism in Lymphoma Patients Undergoing Chemotherapy and its Clinical Significance.

This study investigated the risk factors of thromboembolism (TE) in lymphoma patients undergoing chemotherapy and its clinical significance. A total of 304 lymphoma patients who received chemotherapy from January 2012 to July 2019 were retrospectively analyzed, including 111 patients with and 193 patients without TE. The clinical characteristics and related laboratory test results were compared between the 2 groups using univariate analysis, while the risk factors for TE in lymphoma patients undergoing chemotherapy were analyzed using multivariate logistic regression analysis. Univariate analysis revealed an increase in the risk of TE among lymphoma patients with chemotherapy in the following categories: female patients, patients with body mass index <18.5 or > 24, patients aged ≥60 years, those with platelet abnormality before chemotherapy, single hospital-stay patients, and Ann Arbor stage III/IV patients. Multivariate logistic regression analysis revealed that for platelet count abnormality before chemotherapy, Ann Arbor stage III/IV and female patients represented independent risk factors for TE among lymphoma patients after chemotherapy (P < .05). For lymphoma patients treated with chemotherapy, the risk of TE occurring in women, patients with platelet abnormalities before chemotherapy, and patients at Ann Arbor stage III/IV was significantly higher compared with other patients. For these patients, we recommend prophylactic anticoagulant therapy.

[Clinical Value of Serum Amyloid A and Misfolded Transthyretin for Relapsed/Refractory Diffuse Large B-Cell Lymphoma Patients].

OBJECTIVE: To evaluate the clinical value of serum amyloid A (SAA1/2) and misfolded transthyretin (TTR) for relapsed/refractory diffuse large B-cell lymphoma (R/R DLBCL) patients. METHODS: 30 R/R DLBCL patients were enrolled as observation group, 20 remission/stabilization DLBCL and 10 chronic lymphadenitis patients were enrolled as control group. SELDI technique, Tris-Tricine sodium dodecyl sulfate-polyacrylamide gel electro-phoresis, the shotgun-LTQ-MS method, and bioinformatics technique were used to detected and analyzed SAA and TTR in R/R DLBCL patients. SPSS 21.0 software was used to analyze the relationship between the high expression of SAA, misfolded TTR in serum and the clinicopathological features, survival time of R/R DLBCL. patients Chi-square test was used to analyze clinical count data, Kaplan-Meier curve was used for survival analysis, and Log-Rank test was used to compare single-factor survival differences. RESULTS: The high expression of SAA and TTR (SAA+TTR+) was significantly associated with extranodal lesion, high level of LDH, and NCCN-IPI scores, and also correlated with non-GCB type. TTR+ was correlated with C-MYC in pathological tissue, while SAA+ was also associated with B-symptoms. The survival time of patients in SAA+, TTR+, and SAA+TTR+ group were shorter than that in control group. CONCLUSION: Both SAA and misfolded TTR are poor prognosis factors of R/R DLBCL patients.

[Effect of P53 Expression on Prognosis of Patients with Double Expressor Lymphoma].

OBJECTIVE: To investigate the effect of P53 expression on prognosis of patients with double expressor lymphoma(DEL) and the interaction between the expression of MYC, BCL2 and P53 in diffuse large B-cell lymphoma(DLBCL). METHODS: Eighty-eight patients with newly diagnosed DLBCL from 1st September 2012 to 31th May 2018 in Shanxi Dayi Hospital affiliated to Shanxi Medical University were selected. The expressions of MYC、BCL2、P53、CD10、BCL6、MUM and Ki-67 were tested by immunohistochemistry method. The overall survival of patients was analyzed by the Kaplan-Meier method and compared by the log-rank test. The prognostic effect of MYC, BCL2 and P53 expression was analyzed by univariate and multivariate analysis. RESULTS: Compared with patients without P53 expression, the patients with P53 expression had higher LDH level, higher NCCN-IPI scores, lower response to chemotherapy，poorer overall survival(OS) and a higher rate of death(P<0.05). In patients who had diffuse large B-cell lymphoma associated with MYC, BCL2 expression or MYC+/BCL2+ double expression， compared with the patients whom without P53 expression, P53 expression associated with a significant worse OS (P<0.05). The patients with concurrent MYC and P53 expression had a worse OS, compared with patients with either P53 or MYC expression(P<0.05). In patients with MYC+/P53+ co-expression, BCL2 expression did not correlate with poorer survival significantly(P>0.05). Among lymphoma patients with MYC+/P53+, MYC+/BCL2+ and BCL2+/P53+ co-expression, the patients with MYC+/P53+ co-expression had the worse OS (3 year OS rate：31.6%), followed by the subgroup of patients with MYC-/BCL2+/P53+(3 year OS rate：46.2%), patients with MYC+/BCL2+/P53- expression(3 year OS rate： 636%) showed a longer OS compared with the other two subgroups（P<0.05）. Multivariate analysis demonstrated that P53 expression and NCCN-IPI were independent prognostic factors in this patient cohort. CONCLUSION: P53 and MYC expressions have a synergistically negative prognostic effect in DLBCL patients. P53 expression augments the negative prognostic effect of MYC+/BCL2+ double expression. Patients with MYC+/P53+ co-expression have a worse prognosis in comparison with the patients with MYC+/BCL2+ double expression.

Clinicopathologic Significance and Prognostic Value of Programmed Cell Death Ligand 1 (PD-L1) in Patients With Hepatocellular Carcinoma: A Meta-Analysis.

Background: There is still a dispute over an issue of the clinical pathology and prognostic of programmed cell death ligand 1 (PD-L1) in hepatocellular carcinoma (HCC) patients. Here, we undertook this meta-analysis to survey the conceivable role of PD-L1 in HCC. Method: We searched databases like MEDLINE, EMBASE, and Google Scholar for relevant studies published in English up to February 13, 2018. We implemented the appraisal of the eligible studies according to the choice criterion. We used Hazard ratio (HR) and its 95% confidence interval (95% CI) to evaluate the prognostic role of PD-L1 for overall survival (OS), disease-free survival (DFS), and recurrence-free survival (RFS). Odds ratio (OR) and the corresponding 95% CI were calculated to evaluate the connection between PD-L1 and clinicopathological features. Publication bias was tested. Results: 13 studies, which published range from 2009 to 2017 were contained in this meta-analysis, involving 1,843 patients with HCC. The results indicated that high PD-L1 could predict shorter OS (HR = 1.57, 95% CI: 1.09-2.27, P < 0.00001) as well as poorer DFS (HR = 2.07, 95% CI: 1.20-3.58, P = 0.009). Additionally, high PD-L1 expression was correlated to liver cirrhosis (OR = 1.66, 95% CI: 1.10-2.50, P = 0.02), poorer tumor Barcelona Clinical Liver Cancer (BCLC) stage (OR = 0.30, 95% CI: 0.10-0.88, P = 0.03) and portal vein invasion (OR = 1.96, 95% CI: 1.04-3.68, P = 0.04), but had no correlation with age, gender, tumor size, number of tumors, AFP, vascular invasion, HBVs-Ag, Anti-HCV, differentiation or TNM stage. Besides, no significant publication bias was found among these identified studies. Conclusion: The meta-analysis suggested that PD-L1 overexpression could foresee worse OS and DFS in HCC. Moreover, the PD-L1 expression has to bear on liver cirrhosis, portal vein invasion, and BCLC stage.

Immunotherapy of DC-CIK cells enhances the efficacy of chemotherapy for solid cancer: a meta-analysis of randomized controlled trials in Chinese patients.

OBJECTIVE: Professional antigen-presenting dendritic cells (DCs) and cytokine-induced killer (CIK) cells, components of anti-cancer therapy, have shown clinical benefits and potential to overcome chemotherapeutic resistance. To evaluate whether DC-CIK cell-based therapy improves the clinical efficacy of chemotherapy, we reviewed the literature on DC-CIK cells and meta-analyzed randomized controlled trials (RCTs). METHODS: We searched several databases and selected studies using predefined criteria. RCTs that applied chemotherapy with and without DC-CIK cells separately in two groups were included. Odds ratio (OR) and mean difference (MD) were reported to measure the pooled effect. RESULTS: Twelve reported RCTs (826 patients), which were all performed on Chinese patients, were included. Combination therapy exhibited better data than chemotherapy: 1-year overall survival (OS) (OR=0.22, P<0.01), 2-year OS (OR=0.28, P<0.01), 3-year OS (OR=0.41, P<0.01), 1-year disease-free survival (DFS) (OR=0.16, P<0.05), 3-year DFS (OR=0.32, P<0.01), objective response rate (ORR) (OR=0.54, P<0.01), and disease control rate (DCR) (OR=0.46, P<0.01). Moreover, the levels of CD3(+) T-lymphocytes (MD=-11.65, P<0.05) and CD4(+) T-lymphocytes (MD=-8.18, P<0.01) of the combination group were higher. CONCLUSIONS: Immunotherapy of DC-CIK cells may enhance the efficacy of chemotherapy on solid cancer and induces no specific side effect. Further RCTs with no publishing bias should be designed to confirm the immunotherapeutic effects of DC-CIK cells.