Construction and validation of risk model of EMT-related prognostic genes for kidney renal clear cell carcinoma.

BACKGROUND: Kidney renal clear cell carcinoma (KIRC) is a prevalent type of urological malignancy. The present study aimed to predict biomarkers for KIRC. METHODS: We collected transcriptomic and clinical information for KIRC from The Cancer Genome Atlas and GSE22541 cohorts. RESULTS: Unsupervised clustering of 35 epithelial-mesenchymal transformation (EMT)-related differentially expressed gene profiles divided samples into two clusters with distinct immune characteristics. Six genes (IL20RB, DDC, ANKRD36BP2, F2RL1, TEK, and AMN) were found to construct a prognostic risk model using multivariate Cox regression analysis. Kaplan-Meier analysis suggested the better prognosis of the KIRC patients in the low-risk group than that in the high-risk group. Immune infiltration analyses was conducted using xCell and single-sample gene set enrichment analysis, indicating that the risk score was associated with the immune microenvironment of the KIRC. Prognostic marker gene-targeted medications with high drug sensitivity were predicted in KIRC patients. CONCLUSIONS: In summary, the present study identified IL20RB, DDC, ANKRD36BP2, F2RL1, TEK, and AMN as prognostic biomarkers, providing insight into immunotherapy and gene-targeted drugs of KIRC.

[Expression and Clinical Significance of Exosome Component 4 in Newly Diagnosed Patients with Diffuse Large B-Cell Lymphoma].

OBJECTIVE: To explore the expression of Exosome Component 4（EXOSC4） in the tissues of newly diagnosed patients with diffuse large B-cell lymphoma (DLBCL) and its clinical significance. METHODS: The expression of EXOSC4 protein in the tissues of 181 newly diagnosed DLBCL patients was analyzed by immunohistochemical staining. Clinical data were collected. The correlation between EXOSC4 protein expression in the tissues of newly diagnosed DLBCL patients and clinical features were analyzed and its prognostic significance. RESULTS: The positive rate of EXOSC4 protein expression was 68.51% in the tissues of 181 newly diagnosed DLBCL patients. These patients were divided into two groups, with 44 cases in high expression group and 137 cases in low expression group. There were no significant differences in age, gender, B symptoms, serum lactate dehydrogenase (LDH) level, Eastern Cooperative Oncology Group (ECOG) score, Ann Arbor stage, extranodal disease, International Prognostic Index (IPI) score, National Comprehensive Cancer Network IPI (NCCN-IPI) score, and cell origin between the two groups (P>0.05). Cox multivariate regression analysis showed that high EXOSC4 protein expression in tissues was an independent poor prognostic factor for OS and PFS in newly diagnosed DLBCL patients (all P<0.05). K-M survival analysis showed that newly diagnosed DLBCL patients with high EXOSC4 protein expression had significantly shorter overall survival (OS) and progression free survival (PFS) than those patients with low EXOSC4 protein expression (all P<0.05). CONCLUSION: High EXOSC4 protein expression in tissues of newly diagnosed DLBCL patients is an independent poor prognostic factor for survival.

[Release of Exosomes Derived from Leukocyte-Depleted Red Cell Suspension and Its Regulation on Hematological Tumor Cells].

OBJECTIVE: To investigate the release of exosome (Exo) from leukocyte-depleted red cell suspension (LDRCS) at different storage time and its regulation on proliferation of hematological tumor cells and possible mechanism. METHODS: The Exo (RBC-Exo) in LDRCS at different storage time was obtained by ultracentrifugation, and the morphology and immunological marker of RBC-Exo were detected by transmission electron microscopy and Western blot, respectively. The particle size distribution of RBC-Exo in LDRCS at different storage time was detected by Dynamic Light Scattering. CCK-8 assay was used to explore the effect of RBC-Exo on hematological tumor cell proliferation. Western blot was used to detect the expression of proliferation-related proteins in hematological tumor cells after co-culture with RBC-Exo. RESULTS: RBC-Exo was isolated, which was characterized by cup-like shape, particle size distribution ranged from 20 to 200 nm, CD63/TSG101 enriched, Calnexin negative, CD235a positive and CD41 negative. The particle size distribution of RBC-Exo from LDRCS between middle was not significantly different and late stored stage. But the particle size distribution of RBC-Exo at middle-late stored stage(>14 d) was larger than that at early stored stage (≤14 days). Compared with the control group, RBC-Exo could significantly promote the proliferation of HBL1, U2932 and Jurkat cells. Compared with the control group, the cycle-related protein P21 was significantly down-regulated in HBL1, U2932 and Jurkat cells after co-culture with RBC-Exo for 3 days, while the anti-apoptotic protein BCL-2 was not changed significantly. CONCLUSION: The morphology of RBC-Exo from LDRCS at middle-late stored stage was different from that at early stored stage. RBC-Exo could promote the proliferation of hematological tumor cells, possibly by regulating the expression of cycle-associated protein P21.

miR-638 represses the stem cell characteristics of breast cancer cells by targeting E2F2.

OBJECTIVE: The miR-638 acted as a tumor suppressor and E2F transcription factor 2 (E2F2) was a critical regulator in some cancers, while the role of them on stemness of breast cancer stem cells (BCSCs) was rarely detailed. Hence, we focused on exploring the effects of miR-638 and E2F2 on BCSCs stemness. METHODS: The proportion of CD24 -/CD44 + cells of BCSCs was detected by flow cytometry. The target relationship of miR-638 and E2F2 was explored using luciferase assays. The ability of self-renewal, proliferation, and invasion of BCSCs were determined by Mammosphere forming, Cell Counting Kit-8 (CCK-8), colony formation, and transwell assays. Xenograft tumor was established to detect the influence of miR-638 on tumor growth. RESULTS: miR-638 was down-regulated, while E2F2 was elevated in breast cancer. The E2F2 level was negatively correlated with miR-638. The BCSCs represented higher proportion of CD24 -/CD44 + cells and levels of sex determining region Y-box 2 (SOX2) and octamer-binding transcription factor 4 (OCT4). The miR-638 was down-regulated and E2F2 was increased in BCSCs. MiR-638 could target to E2F2 and decreased the level of E2F2 in BCSCs cells. Overexpression of miR-638 decreased the proportion of CD24 -/CD44 + cells and the levels of SOX2 and OCT4 by inhibiting E2F2. The overexpression of miR-638 also inhibited the abilities of self-renewal, proliferation, and invasion of BCSCs by inhibiting E2F2. The miR-638 overexpression inhibited the breast tumor growth. CONCLUSION: MiR-638 represses the characteristics and behaviors of BCSCs by targeting E2F2. MiR-638 may be a potential target for breast cancer therapy.

[Analysis of the efficiency and influence factors of PBSC collection with AutoPBSC and MNC procedure of cell separator].

This study was aimed to analyze the efficiency and influence factors of PBSC collection by an automatic (AutoPBSC procedure) and a semiautomatic apheresis procedure ( MNC procedure) of COBE Spectra cell separators. According to the different objects, A total of 109 apheresis cases were divided into autologous cohort (patient) and allogeneic cohort (donor). The quantity and quality of the collections and the characteristics of apheresis procedure were compared, the yields and influence factors of two cohorts with two kinds of procedures were analyzed respectively. The results showed that the collections of two procedure in patients and donors which processed the similar blood volumes were insignificantly different in MNC%, CD34⁺ %, CD34⁺ cell counts and Hb concentration (P > 0.05) ; the collections by AutoPBSC procedure had got fewer platelets, less product volumes whereas more ACD-A used, longer apheresis time in comparison with MNC procedure (P < 0.05). Correlation analysis indicated that MNC (r = 0.314,P = 0.015) , CD34⁺ cell counts (r = 0.922, P = 0.000) in collections were positively correlated with preahperesis in the autologus cohort by two procedures, CD34⁺ cell counts were correlated with WBC (r = 0.369, P = 0.004) and MNC (r = 0.495,P = 0.000) in collections; MNC (r = 0.896, P = 0.000) was positive correlated with preahperesis by AutoPBSC procedures and CD34⁺ cell counts also (r = 0.666,P = 0.000) by MNC procedure in the allogeneic cohort. Male had got more MNC and CD34⁺ cell counts than female (P < 0.05), age ≤ 40 had got more MNC and CD34⁺ cell counts than age>40 (P < 0.05) in patients by AutoPBSC procedure; age > 40 had got more CD34⁺ cell counts than age ≤ 40 by MNC procedure(P < 0.05). Only male had got more MNC and CD34⁺ cell counts than female (P < 0.05) by MNC procedure in donors. It is concluded that with same amount of blood processing, the PBSC collections from autologous patients and allogeneic donors had got a high degree of uniformly in purity of MNC and purity and concentration of CD34(+) cell counts by two procedure, whereas sex and age imposed more influence on PBSC collection in autologous.

[Clinical study on unilateral pedicle screw fixation and interbody fusion for the treatment of lumbar degenerative diseases under Quadrant system].

OBJECTIVE: To compare the clinical effects of unilateral pedicle screw fixation (uni-PS) assisted by Quadrant system and bilateral pedicle screw fixation (bi-PS) for the treatment of lumbar degenerative diseases. METHODS: From October 2008 to October 2010,102 patients with lower back pain, unilateral lower limb radiating pain or paraesthesia were treated with pedicle screw fixation and lumbar interbody fusion. There were 67 males and 35 females with an average age of 51.5 years ranging from 34 to 69 years. The patients were randomly divided into two groups (group A and group B) according to the internal fixation type. The patients of group A (n=50) underwent with minimally transforaminal lumbar interbody fusion (TLIF) and unilateral pedicle screw fixation with one single cage placement assisted by Quadrant system;and the patients of group B (n = 52) underwent with posterior lumbar interbody fusion (PLIF) and bilateral pedicle screw fixation with one single cage placement. There were no significant differences between two groups in general information (P > 0.05). VAS score and ODI score system were used to evaluate the preoperative and postoperative pain and function recovery. Operative time, volume of blood loss, fusion rate and complication rate were compared and analyzed by statistical test. RESULTS: All the patients were followed up from 12 to 21 months with an average of 18.2 months. In the group A,operative time and volume of blood loss were (87.6 +/- 25.5) min and (105.7 +/- 27.2) ml, respectively; VAS score of low back pain and leg pain, ODI score decreased respectively from preoperative 7.2 +/- 1.4, 7.9 +/- 1.1, 42.2 +/- 11.8 to 3.2 +/- 0.6, 3.0 +/- 0.7,15.6 +/- 2.3 at one month after operation; the fusion rate was 96.0% (48/50) and the complication rate was 4.00% (2/50). In the group B,operative time and volume of blood loss were (160.3 +/- 20.5) min and (220.6 +/- 25.5) ml, respectively; VAS score of low back pain and leg pain, ODI score decreased respectively from preoperative 7.3 +/- 1.1, 8.1 +/- 0.9, 43.1 +/- 12.0 to 3.3 +/- 0.4, 3.2 +/- 0.3, 14.9 +/- 2.6; the fusion rate was 96.2% (50/52) and the compli- cation rate was 5.77% (3/52). There were no statistically significant differences between the two groups in fusion rate, complication rate, VAS pain and ODI score. Whereas the operative time and blood loss in group A were significantly lower than that of group B. CONCLUSION: Minimally invasive unilateral pedicle screw fixation is a safe and feasible method for the treatment of lumbar degenerative diseases. It is as effective as the bilateral fixation in lumbar spinal fusion. In addition, it has the advantages of short operative time, less volume of blood loss, high fusion rate, etc.