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The aim of the present study was to use the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR‑associated (Cas) 9‑mediated gene knockout technology for the rapid classification of the differential function of micro (mi)RNAs screened using miRNA expression profiling by microarray. The rational design of single guide RNAs for the CRISPR/Cas9 system was verified to function in human LNCaP cells with rapid and efficient target gene editing. miRNA (miR)‑205, miR‑221, miR‑222, miR‑30c, miR‑224, miR‑455‑3p, miR‑23b and miR‑505 were downregulated in patients with prostate cancer (PCa) and were experimentally validated to function as tumor suppressors in prostate cancer cells, affecting tumor proliferation, invasion and aerobic glycolysis. In addition, the data of the present study suggested that miR‑663a and mfiR‑1225‑5p were upregulated in prostate cancer tissues and cell proliferation of miR‑663a and miR‑1225‑5p knockout PCa cells was significantly lower compared with miR‑NC cells. Furthermore, knockout of miR‑1225‑5p and miR‑663a significantly decreased the lactate production in LNCaP cells in vitro. In conclusion, the present study offered a simple and efficient method for rapidly classifying miRNA function by applying CRISPR/Cas9 in LNCaP cells. The present study suggested, for the first time to the best of the authors' knowledge, that the aberrant expression of miR‑663a and miR‑1225‑5p may be involved with the progression of prostate cancer, implying their potential as candidate markers for this type of cancer. However, the precise role of miR‑663a and miR‑1225‑5p in accelerating the development of prostate cancer and promoting tumor progression remains to be elucidated.

To investigate immune profile consisting of stromal PD-L1 expression, inhibitory or non-T-cell inflamed tumor microenvironment that may predict response to anti-PD-L1/PD-1 immunotherapy in prostate cancer, we validated the specificity of a PD-L1 monoclonal antibody (E1L3N) and identified PD-L1 specific expression in prostatic stromal nerve cells. PD-L1 expression was analyzed in 73 primary prostate cancers and 7 castration-resistant prostate cancers (CRPC) by immunohistochemistry (IHC) and resulting data from primary prostate cancers were correlated with tumor-associated lymphocytes (TALs), clinicopathological characteristics and clinical outcome. PD-L1 was expressed in the tumor cells in only one primary prostate cancer case and none of the CRPC. However, PD-L1 was frequently observed in the nerve branches in the tumor-associated stroma (69 of 73 cases, 94.5%), supported by colocalization with axonal marker PGP9.5. FoxP3-, CD3- and CD8-positive T lymphocytes were observed in 74.6% (47/63), 98.4% (62/63) and 100% (61/61) of the cases, respectively. The density of PD-L1

Discs large homolog 1 (DLG1) belongs to the modular proteins Membrane Associated Guanylate Kinases (MAGUKs) and plays a major role in the formation of cell-cell junctions and cell polarity. DLG1 varies in expression and localization among malignancies. However, the clinical significance of DLG1 in the context of human colorectal cancer (CRC) remains unclear. The purpose of this study study is to determine the association of DLG1 with clinicopathological characteristics and prognosis of CRC patients.DLG1 expression in human CRC was detected by immunohistochemical analysis and validated with mRNA data from a high-throughput sequencing TCGA datasets. The association of DLG1 expression with clinicopathological features in CRC patients was then statistically analyzed.Immunohistochemistry analysis found that DLG1 expression was significantly elevated in CRC tissues, compared with the expression in adjacent non-cancerous colon tissues (These findings suggest that the increased expression of DLG1 may be predictive of an unfavorable prognosis in CRC patient and serve as a novel therapeutic target in this malignancy.

To investigate the relationship of oxidative stress with DNA integrity and semen parameters in infertile men with varicocele (VC).This prospective study included 98 infertile males with VC. According to the levels of reactive oxygen species (ROS) in the semen, we divided the patients into a high ROS group (n=44) and a low ROS group (n=54), determined the sperm DNA fragmentation index (DFI), motility and morphology, and analyzed their correlation with ROS in the two groups of patients.Compared with the patients of the low ROS group, those of the high ROS group showed a significantly higher DFI (27.38±8.10 vs 34.49±6.05, P=0.039) and a higher concentration of seminal leukocytes (［0.65±0.15］×10⁶/ml vs ［0.86±0.41］×10⁶/ml, P=0.022), but lower sperm motility (［36.16±22.83］% vs ［18.22±25.21］%, P=0.017), percentage of progressively motile sperm (［23.34±11.53］% vs ［16.34±9.22］%, P=0.041), sperm curvilinear velocity (［27.03±6.21］ vs ［20.62±4.38］ μm/s, P=0.013), and sperm linearity (［29.75±8.24］% vs ［18.30±7.93］%, P=0.024). Spearman correlation analysis indicated that the ROS level was correlated positively with the concentration of seminal leukocytes (r=0.41, P0.01) and DFI (r=0.21, P=0.006), but negatively with sperm curvilinear velocity (r=－0.24, P=0.017), linearity (r=－0.24, P=0.021), motility (r=－0.31, P=0.002), and the percentage of progressively motile sperm (r=－0.41, P=0.012). Additionally, the sperm DFI manifested a significant negative correlation with sperm motility (r=－0.29, P0.01) and the percentage of progressively motile sperm (r=－0.34, P0.01).The level of seminal ROS is positively correlated with the sperm DFI in infertile men with varicocele, and both the ROS level and DNA integrity are associated with semen parameters.目的：分析精索静脉曲张(VC)不育患者氧化应激同DNA完整性及精液参数的关系。方法：采用前瞻性研究，纳入98例患者，根据活性氧水平（ROS)将VC不育患者分为ROS低水平组(54例)及高水平组（44例），分析两组精子DNA碎片指数（DFI）、精子活力、形态等参数的差异，并分析它们之间的相关性。结果： ROS高水平组DFI显著高于ROS低水平组［（34.49±6.05)% vs (27.38±8.10)%， P=0.039］。与ROS低水平组比较，ROS高水平组精子活力［（25.21±18.22)% vs (36.16±22.83)%， P=0.017］、前向运动精子百分率［（16.34±9.22)% vs (23.34±11.53)%，P=0.041］、曲线速率［（20.62±4.38)% vs (27.03±6.21)%，P=0.013］及直线性率［（18.30±7.93)% vs (29.75±8.24)%， P=0.024］均显著降低，而精液白细胞数显著升高［（0.86±0.41 )×10⁶/ml vs (0.65±0.15)×10⁶/ml ， P=0.022］。Spearman相关性分析表明ROS水平同精液白细胞数（r=0.41，P0.01）及DFI（r=0.21，P=0.006）呈显著正相关，同曲线速率（r=－0.24，P=0.017）、直线性率（r=－0.24，P=0.021）、精子活力（r=－0.31，P=0.002）及前向运动精子百分率（r=－0.41，P=0.012）呈显著负相关。DFI同精子活力（r=－0.29，P0.01）、前向运动精子百分率（r=－0.34，P0.01）呈显著负相关。结论：精浆ROS水平同VC不育患者DFI显著正相关。氧化应激及DNA完整性可影响患者的精子参数。.

The global physiological function of specifically expressed genes of prostate cancer in Chinese patients is unclear. This study aims to determine the genome-wide expression of genes related to prostate cancer in the Chinese population. Genes that were differentially expressed in prostate cancer were identified using DNA microarray technology. Expressions were validated by using real-time PCR. The identified genes were analyzed using the ingenuity pathway analysis (IPA) to investigate the gene ontology, functional pathway and network. A total of 1,444 genes (Fold time ≥ 1.5; P ≤ 0.05) were differentially expressed in prostate primary tumor tissue compared with benign tissue. IPA revealed a unique landscape where inductions of certain pathways were involved in Cell Cycle Regulation and proliferation. Network analysis not only confirmed that protein interactions lead to the deregulation of DNA Replication, Recombination and Repair, Cellular Compromise and Cell Cycle, Genetic Disorders and Connective Tissue Disorders, but it was also observed that many of the genes regulated by Myc contributed to the modulation of lipid Metabolism and Nucleic Acid Metabolism. Both pathway and network analysis exhibited some remarkable characteristics of prostate cancer for Chinese patients, which showed profound differences from that of other non-Chinese populations. These differences may provide new insights into the molecular cascade of prostate cancer that occurs in Chinese patients.

Our previous microarray data showed that microRNA-224 (miR-224) was downregulated in human prostate cancer (PCa) tissues compared with adjacent benign tissues. However, the underlying mechanisms by which miR-224 is involved in PCa remain unclear. In this study, we identified TRIB1 as a target gene of miR-224. Forced expression of miR-224 suppressed PCa cell proliferation, invasion and migration, and promoted cell apoptosis by downregulating TRIB1. Moreover, the expression level of miR-224 in PCa tissues was negatively correlated with that of TRIB1. miR-224 downregulation was frequently found in PCa tissues with metastasis, higher PSA level and clinical stage, whereas TRIB1 upregulation was significantly associated with metastasis. Both miR-224 downregulation and TRIB1 upregulation were significantly associated with poor biochemical recurrence-free survival of patients with PCa. In conclusion, these findings reveal that the aberrant expression of miR-224 and TRIB1 may promote PCa progression and have potentials to serve as novel biomarkers for PCa prognosis.

Background Our recent study showed the global physiological function of the differentially expressed genes of prostate cancer in Chinese patients was different from that of other non-Chinese populations. microRNA are estimated to regulate the expression of greater than 60% of all protein-coding genes. To further investigate the global association between the transcript abundance of miRNAs and their target mRNAs in Chinese patients, we used microRNA microarray approach combined with bioinformatics and clinical-pathological assay to investigate the miRNA profile and evaluate the potential of miRNAs as diagnostic and prognostic markers in Chinese patients. Results A total of 28 miRNAs (fold change ≥1.5; P ≤ 0.05) were differentially expressed between tumor tissue and adjacent benign tissue of 4 prostate cancer patients.10 top Differentially expressed miRNAs were validated by qRT-PCR using all 20 tissue pairs. Compared to the miRNA profile of non-Chinese populations, the current study showed that miR-23b, miR-220, miR-221, miR-222, and miR-205 maybe common critical therapeutic targets in different populations. The integrated analysis for mRNA microarray and miRNA microarray showed the effects of specifically inhibiting and/or enhancing the function of miRNAs on the gene transcription level. The current studies also identified 15 specific expressed miRNAs in Chinese patients. The clinical feature statistics revealed that miR-374b and miR-19a have significant correlations with clinical-pathological features in Chinese patients. Conclusions Our findings showed Chinese prostate cancer patients have a common and specific miRNA expression profile compared with non-Chinese populations. The miR-374b is down-regulated in prostate cancer tissue, and it can be identified as an independent predictor of biochemical recurrence-free survival.