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Background. Kidney renal clear cell carcinoma (KIRC) lacks effective prognostic biomarkers and the role and mechanism of N6-methyladenosine (m6A) modification of long noncoding RNAs (lncRNAs) in KIRC remain unclear. Methods. We extracted standard mRNA-sequencing and clinical data from the TCGA database. The prognostic risk model was obtained by Lasso regression and Cox regression. We randomly divided the samples into training and test sets, each taking half of the cases. Based on Lasso regression and Cox regression for training set, the prognostic risk signature was constructed; risk scores were calculated with the R package “glmnet.” Based on the median value of the prognostic risk score, risk scores were calculated for each patient and we divided all KIRC samples into high-risk and low-risk groups. Then, high- and low-risk subtypes were established and their prognosis, clinical features, and immune infiltration microenvironment were evaluated in test set and the entire sampled data set. The reliability of the prognostic model was confirmed by receiver operating characteristic curve analysis. Results. We found 28 prognostic m6A-related lncRNAs and established a m6A-related lncRNAs prognostic signature. Risk score = A C 015813.1 ∗ 0.0086 + E M X 2 O S ∗ − 0.0101 + L I N C 00173 ∗ 0.0309 + P W A R 5 ∗ − 0.0146 + S N H G 1 ∗ 0.0043 . The signature showed a better predictive ability than other clinical indicators, including tumor node metastasis classification (TNM), histological, and pathological stages. In the high-risk group, M0 macrophages, CD8+ T cells, and regulatory T cells had significantly higher scores. Contrarily, in the low-risk group, activated dendritic cells, M1 macrophages, mast resting cells, and monocytes had significantly higher scores. In the high-risk group, LSECtin was overexpressed. In the low-risk group, PD-L1 was overexpressed. Moreover, high-risk patients may benefit more from AZ628. Conclusions. In conclusion, prognosis prediction of patients with KIRC and new insights for immunotherapy are provided by the m6A-related lncRNA prognostic signature.

Introduction: T-cell immunoglobulin-3 (Tim-3) antibody drugs can treat malignant renal tumors but are expensive. To overcome this limitation, Lactococcus lactis host bacteria were used to express Tim-3 single-chain antibodies. Methods: The pLAN-CTB-Tim3scFv plasmid was constructed using molecular cloning technology and transformed into Lactococcus lactis. Expression and immune activity of proteins in the transformed bacteria were analyzed using Western blotting and enzyme-linked immunosorbent assay in vitro. A mouse subcutaneously transplanted tumor model of renal adenocarcinoma was constructed. The promoting effect of transformed bacteria on mouse spleen lymphocyte activation and their inhibitory effect on transplanted tumors were analyzed. Results: Transformed L. lactis NZ-CTB-Tim3scFv and NZ-Tim3scFv were successfully constructed. CTB-Tim3scFv secreted by NZ-CTB-Tim3scFv showed immunological activity. Compared with the NZ-Tim3scFv and NZ-Vector groups, the subgroups of splenic lymphocytes in the NZ-CTB-Tim3scFv group had a higher proportion of CD3+CD4+, CD3+CD8a+, and CD3+CD69+ cells. Ki67 and CD31 expression in the NZ-CTB-Tim3scFv group was significantly reduced. Tumor volume in the NZ-CTB-Tim3scFv group increased the least. Discussion/Conclusion: Secretion of CTB-Tim3scFv promoted the proliferation and activation of spleen lymphocytes and inhibited growth, cell proliferation, and angiogenesis of tumors. The proposed method is low cost and convenient with potential to become a new immunotherapy approach for renal-cell carcinoma.

Sunitinib, an inhibitor of receptor tyrosine kinase, possesses anti-tumor activity in renal cell carcinoma (RCC) through its anti-angiogenic effects. However, patients with advanced RCC are resistant to sunitinib. Dysregulated circRNAs has been shown to be associated with drug resistance in various tumors. However, little is known about the effect of circRNA_001895 on sunitinib resistance of RCC. First, the expression of circRNA_001895 was found to be higher in sunitinib-resistant RCC tissues than chemosensitive tumor tissues. Half maximal inhibitory concentration of sunitinib-resistant RCC cells (786-O/R and ACHN/R) was higher than sunitinib-sensitive 786-O and ACHN cells. CircRNA_001895 was also upregulated in 786-O/R and ACHN/R cells. Second, data from colony formation and flow cytometry analysis showed that knockdown of circRNA_001895 suppressed cell proliferation and promoted cell apoptosis in 786-O/R and ACHN/R cells. Moreover, the protein expression of phosphorylated histone H2AX (γH2AX) was enhanced, while phosphorylated DNA-dependent protein kinase (p-DNA-PK) and Rad51 were reduced in 786-/R and ACHN/R by knockdown of circRNA_001895. Lastly, knockdown of circRNA_001895 conferred sensitivity of

Background T cell immunoglobulin-3(Tim-3) is an immune checkpoint molecule; Tim-3 antibody is suitable for treating malignant renal tumours. However, Tim-3 antibody drugs are expensive, which limits their application. To overcome the disadvantages of expensive immunotherapeutic drugs, Lactococcus lactis was used as the host bacteria to express Tim-3 single-chain antibody in the intestine, and its promotion of the mouse immune system and inhibitory effects on the transplanted tumour of kidney cancer in mice were tested. Methods Molecular cloning technology was used to construct plasmids pLAN-CTB-Tim3scFv and pLAN-Tim3scFv, which were transformed into Lactococcus lactis. The expression of the transformed bacteria was analysed using western blotting, and the immune activity of secreted proteins of the transformed bacteria was detected using ELISA in vitro. A subcutaneous transplanted tumour model of renal adenocarcinoma was constructed in RAG mice, and the promoting effect of transforming bacteria on the activation of mouse spleen lymphocytes, and its inhibitory effect on transplanted tumours in mice was analysed. Results (1) Transformed Lactococcus lactis NZ -CTB-Tim3scFv and NZ -Tim3scFv, which secrete CTB-Tim3scFv and Tim3scFv single-chain antibodies, were successfully constructed. (2) CTB-Tim3scFv secreted by NZ-CTB-Tim3scFv transformed bacteria showed immunological activity. (3) Compared with the NZ-Tim3scFv and NZ-Vector groups, the characteristics of the subgroups of splenic lymphocytes in the NZ-CTB-Tim3scFv group had a higher proportion of CD3+CD4+, CD3+CD8a+, and CD3+CD69+ cells. Ki67 and CD31 expression in the NZ-CTB-Tim3scFv group was significantly reduced. The tumour volume of the NZ-CTB-Tim3scFv group increased the least, and was statistically different from that of the other two groups. Conclusions The Tim-3 single-chain antibody gene was successfully constructed and transformed in Lactococcus lactis. After feeding mice NZ-CTB-Tim3scFv transforming Lactobacillus, the CTB-Tim3scFv secreted by the transforming Lactobacillus promoted the proliferation and activation of spleen lymphocytes and inhibited volume growth, cell proliferation, and angiogenesis of the tumour in mice. In summary, apply transgenic lactobacillus secreting CTB-Tim-3scFv, to perform its role in anti-tumor immunotherapy through oral approach, is low cost and convenient. It is expected to become a new way of immunotherapy for renal cell carcinoma.

With the aid of next-generation sequencing technology, pseudogenes have been widely recognized as functional regulators in the development and progression of certain diseases, especially cancer. Our present study aimed to investigate the functions and molecular mechanisms of HSPB1-associated protein 1 pseudogene 1 (HSPB1P1) in renal cell carcinoma (RCC). HSPB1P1 expression at the mRNA levels was determined by quantitative real-time polymerase chain reaction, and its clinical significance was assessed. Cell viability was detected by Cell Counting Kit-8 assay. Cell migration and invasion were detected by transwell assays. The location of HSPB1P1 in RCC cells was detected by subcellular distribution analysis. The direct relationship between HSPB1P1 and miR-296-5p/HMGA1 axis was verified by dual-luciferase reporter assay and RNA immunoprecipitation assay. Our results identify the elevated expression of HSPB1P1 in RCC tissues and cell lines, which predicted advanced progression and poor prognosis in patients with RCC. Knockdown of HSPB1P1 suppressed cell proliferation, migration, and invasion, and reversed epithelial-mesenchymal transition process in RCC. HSPB1P1 was mostly enriched in the cytoplasm and functioned as a miRNA sponge for miR-296-5p and then regulated high-mobility group A1 expression. In conclusion, our study indicated that HSPB1P1 contributed to RCC progression by targeting the miR-296-5p/HMGA1 axis, and should be considered as a promising biomarker and therapeutic target for clinical applications.

Background Urinary microbiota is associated with the recurrence of bladder cancer, but the underlying mechanism remains unclear. The notion that microbiota can upregulate PD-L1 expression in tumors to promote immune escape have been demonstrated. We hypothesized that the urinary microbiota may be involved in the recurrence and progression of non-muscle invasive bladder cancer (NMIBC) by upregulating the PD-L1 expression. For proving this hypothesis, we firstly performed this study to characterize the potential urinary microbial community possibly associated with PD-L1 expression in male patients with NMIBC. Results The subjects (aged 43–79 years) based on their PD-L1 immunohistochemical results were divided into PD-L1-positive group (P group) and PD-L1-negative group (N group) respectively. We observed that P group exhibited higher species richness and diversity (based on Observed species and Ace index, both P P Leptotrichia, Roseomonas, and Propionibacterium) and decrease of some bacterial genera (e.g.,Prevotella and Massilia) were observed in P group as compared with N group. These findings indicate that these genera may affect the expression of PD-L1 through some mechanisms to be studied. PICRUSt analysis showed that several pathways involved in the metabolism of chemical compounds and immune-related disease were enriched in the PD-L1 negative group. Conclusion Our data indicate that urinary microbiota may be an important determinant of PD-L1 expression in male NMIBC patients. The findings of our study may facilitate subsequent study on the role and mechanism of urinary microbiota in the recurrence of NMIBC and may also pave a new way for the better application of PD1 or PD-L1 blockers in bladder cancer in the future.

The formimidoyltransferase cyclodeaminase (FTCD) gene encodes an enzyme required for the catabolism of histidine and tetrahydrofolate (THF). Previous studies showed that FTCD plays a role as a tumour suppressor gene in hepatocellular carcinoma (HCC). It is unknown whether the restoration of functional FTCD may exhibit an anti-tumour effect on HCC. This study constructed a delivery system based on hollow mesoporous organosilica nanotheranostics (HMON) capable of efficiently loading Mn ions and FTCD plasmids. This study showed that the Mn-doped and FTCD-loaded nanoparticles (HMON@Mn-PEI@FTCD) could efficiently induce the expression of FTCD and achieve enhanced magnetic resonance imaging. In vitro results demonstrated that the upregulation of FTCD induced by HMON@Mn-PEI@FTCD nanoparticles dramatically reduced intracellular THF levels, inhibited of NADPH/NADP+ and GSH/GSSG ratios, and induced reactive oxygen species generation and mitochondrial oxidative stress. As a result, cytochrome c release increased with the opening of the mitochondrial permeability transition pore, which finally activated the caspase-dependent cell apoptosis pathway. Therefore, our designed HMON@Mn-PEI@FTCD could induce apoptosis by activating the mitochondria-mediated apoptosis signalling pathway, and finally significantly suppressed the proliferation of HCC both in vitro and in vivo, which provides an effective strategy for the treatment of HCC.

To describe a clinical staging method linked to stepwise treatment indications for ketamine-associated urinary dysfunction (KAUD) based on review of our experience in management of KAUD patients and analysis of their clinical features. The eighty-one KAUD patients hospitalized from January 2008 to June 2014 were studied retrospectively. According to ketamine history, renal and liver function, bladder change and up urinary tract involvement, patients were categorized into a described model of three stages. Discriminant analysis was applied to validate the model. The void volume, micturition interval, nocturnal void frequency and pelvic pain and urgency/frequency (PUF) questionnaire score were, respectively, compared after treatments. There were, respectively, 24, 47 and 10 patients in three stages. The duration of abuse varied (p = 0.047) correlated with clinical stages (p = 0.015, r = 0.268). The severity of LUTS was not significant. The creatinine, estimated glomerular filtration rate and liver function were worse in higher stages (p

To summarize the imaging features of urinary dysfunction associated with ketamine abuse (KAUD) for imaging diagnosis of KAUD.We analyzed the imaging findings in 45 patients with KAUD, all having a history of ketamine abuse and presenting with severe lower urinary tract symptoms. The patients underwent imaging examinations with ultrasonography (n=45), X-ray (n=38), computed tomography (n=28), magnetic resonance imaging (n=10) or single photon emission computed tomography (n=25), and the results were classified and evaluated to identify the common imaging findings.The imaging changes of KAUD were found primarily in the urinary and biliary system. The most common imaging characteristics included thickening of the bladder wall, contracture and decreased functional volume of the bladder, dilation of the ureter and hydronephrosis, stricture of the upper ureter, renal impairment, dilation of the biliary system, and inflammation or swelling of the adjacent organs and lymph nodesKAUD presents with typical imaging changes. Radiologists should be aware of KAUD if the typical imaging features are detected, especially in cases with a history of ketamine abuse.