Effect of the Application of Exosome on Gastric Cancer.

Gastric cancer is one of the most common and highest mortality rate cancers in the world. Exosomes are vesicles secreted by cells carrying different types of molecules, such as protein and RNA. Numerous studies have confirmed that exosomes are involved in various stages of the occurrence and development of gastric cancer and play an important role. With the gradual development, exosomes have been widely employed in the diagnosis and treatment of gastric cancer. In this review, we have provided a basic overview of exosome, and discussed the role of exosome in the occurrence, proliferation, invasion, metastasis, and drug resistance in gastric cancer. In addition, we have emphasized the bright development prospect of exosome in the diagnosis and treatment of gastric cancer. The data on the discovery, diagnosis, treatment, and prognosis of gastric cancer are not particularly optimistic, but the discovery of exosome, applied in diagnosis and treatment, provides a new and effective way to improve the survival rate of patients with gastric cancer.

Long stress-induced non-coding transcript 5: A promising therapeutic target for cancer treatment.

Long non-coding RNAs are RNA molecules with a transcript length of more than 200 nucleotides and without protein-coding ability. They regulate gene expression by interacting with protein, RNA and DNA. Their function is closely related to their subcellular localization, with regulation of gene expression at the epigenetic and transcriptional levels occurring in the nucleus, and at the post-transcriptional and translational levels in the cytoplasm. Long stress-induced non-coding transcript 5 (LSINCT5), which is localized in the nucleus, is overexpressed in many types of cancers such as breast cancer, gastric cancer, ovarian cancer, thyroid cancer, and gastrointestinal cancer. Substantial evidence indicates that there is an obvious connection between cancers and LSINCT5, as it inhibits apoptosis and promotes proliferation, invasion and migration of cancer cells, as well as participates in the pathogenesis and progression of cancer by interacting with DNA, protein and RNA. These findings suggest that LSINCT5 could be a novel biomarker and an emerging therapeutic target in human cancers. In the present study, the structure and corresponding biological function of LSINCT5 were summarized in order to clarify its molecular mechanisms in the progression of various malignant tumors.

The Role of NcRNAs to Regulate Immune Checkpoints in Cancer.

At present, the incidence of cancer is becoming more and more common, but its treatment has always been a problem. Although a small number of cancers can be treated, the recurrence rates are generally high and cannot be completely cured. At present, conventional cancer therapies mainly include chemotherapy and radiotherapy, which are the first-line therapies for most cancer patients, but there are palliatives. Approaches to cancer treatment are not as fast as cancer development. The current cancer treatments have not been effective in stopping the development of cancer, and cancer treatment needs to be imported into new strategies. Non-coding RNAs (ncRNAs) is a hot research topic at present. NcRNAs, which include microRNAs (miRNAs), circular RNAs (circRNAs), and long non-coding RNAs (lncRNAs), participate in all aspects of cancer biology. They are involved in the progression of tumors into a new form, including B-cell lymphoma, glioma, or the parenchymal tumors such as gastric cancer and colon cancer, among others. NcRNAs target various immune checkpoints to affect tumor proliferation, differentiation, and development. This might represent a new strategy for cancer treatment.

Ischemic preconditioning-induced protective effect for promoting angiogenesis in renal ischemia-reperfusion injury by regulating miR-376c-3p/HIF-1α/VEGF axis in male rats.

OBJECTIVE: Ischemic preconditioning (IPC) is defined as a well-established phenomenon in which brief exposure to sublethal episodes of ischemia and reperfusion induces a tolerance to injurious effects of prolonged ischemia by exploiting intrinsic defence mechanisms. The present study was performed to determine the protective effect of IPC on the rat renal ischemia-reperfusion injury (IRI) via miR-376c-3p/HIF-1α/VEGF axis. METHODS: In vivo, these male Sprague-Dawley rats were treated by IRI and IPC. Meanwhile, these rats from different treatment groups were also injected subcutaneously with 2 nmol agomir-376c-3p and/or 10 nmol recombinant rat HIF-1α. At 72 h after reperfusion, serum samples were respectively collected for renal function. Besides, kidney tissues were harvested to observe renal morphology changes. Subsequently, the expression levels of CD31, HIF-1α and VEGF in the kidney tissues were measured using immunohistochemical staining, quantitative real-time PCR, as well as Western blotting analysis at the indicated time points after reperfusion. In vitro, HK-2 cells were used to detect the cell activity by CCK-8 and transfection of mir-376c-3p mimic in Hypoxia/Reoxygenation (H/R) group. RESULTS: In vivo, the pathological changes were significantly relieved in the rats with IPC group, compared to the IRI group. Rats which were treated IPC significantly reduced the levels of blood urea nitrogen (BUN), serum creatinine (Scr) at 24 h after operation, compared to IRI group. After IPC treatment, the expression level of CD31 was obviously decreased, compared to IRI group. A total of differently expressed microRNAs were screened out by microRNA microarray assay in rat renal ischemia tissue, especially showing that miR-376c-3p was selected as the target miRNA. Compared to IRI group, the expression level of miR-376c-3p were obviously higher in IPC-treated group. Double-luciferase reporter assay demonstrated that miR-376c-3p directly targeted HIF-1α. In vitro, IPC significantly increased cell viability of HK-2, and promoted the angiogenesis via up-regulating miR-376c-3p/HIF-1α/VEGF axis. Furthermore, the expression level of HIF-1α was apparently decreased in HK-2 treated with H/R after miR-376c-3p mimic transfection respectively, as well as the increased expression level of VEGF. CONCLUSIONS: Our study provided a novel insight for investigating the protective effect of IPC on renal IRI. Consequently, miR-376c-3p played an important role in renal IRI by promoting angiogenesis via targeting HIF-1α/VEGF pathway in male rats.

RUNX1/miR-582-5p Pathway Regulates the Tumor Progression in Clear Cell Renal Cell Carcinoma by Targeting COL5A1.

Recent evidences indicated that miRNAs played core role in the progression of clear cell renal cell carcinoma (ccRCC). However, its molecular mechanism in ccRCC is still remained unclear. The study was designed to identify the role and regulatory mechanism of miR-582-5p in ccRCC. In this study, the low expression level of miR-582-5p were detected by qRT-PCR in ccRCC patient tumor samples and ccRCC cell lines, respectively. The expression level of miR-582-5p was associated with tumor stage and metastasis. In vivo and in vitro experiments found miR-582-5p inhibit tumor growth via suppressing COL5A1 expression. Additionally, RUNX1 was identified as the negative regulator of miR-582-5p through database prediction and chromatin immunoprecipitation. Finally, the negative relation of RUNX1 and miR-582-5p was verified through rescue experiment both in vitro and in vivo. In summary, miR-582-5p, which was regulated by RUNX1, inhibited tumor growth and invasion by targeting COL5A1, indicating that miR-582-5p may act as a biomarker and that the RUNX1/miR-582-5p/COL5A1 axis could be a potential therapeutic target for ccRCC.

Enzyme-free electrochemical biosensor based on double signal amplification strategy for the ultra-sensitive detection of exosomal microRNAs in biological samples.

Growing evidence suggests that exosomes-encapsulated miRNAs detection is of paramount significance in early diagnostics of cancer due to protection from degradation by ribonuclease. However, exosomal microRNAs with low abundance and subtle variation have restricted their clinical application. Herein, an electrochemical biosensor for highly sensitive detection of exosomal microRNAs has been fabricated based on the double signal amplification strategy. Our proposed amplifier consists of two steps: target miRNA cyclic signal amplification induced by strand displacement reaction (SDR) and subsequent deposition of silver nanoparticles induced by streptavidin-biotin interaction. Consequently, this method shows ultrahigh sensitivity to detect miRNA. Taking miRNA-21 in exosomes as a model analyte, a detection limit of 0.4 fM (S/N = 3) can be obtained. Meanwhile, the method is relatively simple and low-cost without the requirement of enzyme, which has been applied in biological samples successfully. Therefore, our miRNA assay method has shown great promise as molecular tool in the detection of exosomal miRNA and could be widely used in clinic in the future.

A model based on tumor-infiltrating immune cells for predicting the relapse rates of patients with testicular germ cell tumors.

OBJECTIVE: The activities of tumor-infiltrating immune cells (TIICs) play an important role in the outcomes of many types of cancers. Here, we sought to describe the landscape of TIICs in testicular germ cell tumors (TGCT) and to develop a prognostic model based on this information. METHODS: The Cell-type Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT) algorithm was used to determine the proportions of 22 types of TIICs in a TGCT dataset (n = 74). Univariate and multivariate Cox regression analysis were used to develop an immune risk score (IRS) based on the association between TIICs and disease-free survival (DFS). The predictive accuracy of the IRS was evaluated using receiver operating characteristic curves, and the predictive accuracy of a prognostic nomogram was assessed using C-index and calibration curves. The biological functions of IRS-associated genes were evaluated by gene set enrichment analysis. RESULTS: The relative abundances of three TIICs (plasma cells, M2 macrophages, and resting mast cells) were significantly associated with DFS in TGCT patients. In receiver operating characteristic curve analysis, the resulting IRS had areas under the curve of 0.70, 0.793, and 0.827, for predicting 1-, 2-, and 3-year DFS, respectively. Kaplan-Meier analysis confirmed that DFS was shorter for patients with high IRS compared with low IRS. IRS was an independent predictor of disease recurrence (hazard ratio 1.306, 95% confidence interval 1.022-1.668; P = 0.033). The C-index for the nomogram was 0.733. Genes involved in cancer-associated and immunity-associated pathways were enriched in TGCT samples from patients in the high- and low-risk groups, respectively, and expression of four immune checkpoint regulators was significantly lower in the high IRS group compared with the low IRS group. CONCLUSIONS: A TIIC-based IRS may have utility as a complementary tool to predict relapse in patients with TGCT.

Genetic Variant in Long Non-Coding RNA H19 Modulates Its Expression and Predicts Renal Cell Carcinoma Susceptibility and Mortality.

The long non-coding RNA (lncRNA) H19 has been demonstrated to play a crucial role in carcinogenesis, including renal cell carcinoma (RCC). However, the impact of genetic variations in H19 gene on RCC has not been investigated before. In the present study, we sought to evaluate whether genetic polymorphisms in H19 are related to the susceptibility and mortality of RCC. We genotyped four widely studied polymorphisms in H19 and assessed their relationship with susceptibility and prognosis of RCC in a case-control study compromising 1,027 cases and 1,094 controls. The functionality of the important polymorphism was further investigated by real-time polymerase chain reaction and luciferase reporter assay. We found that H19 rs2839698 was significantly associated with risk and prognosis of RCC. Compared with the H19 rs2839698 CC genotype, the variant genotypes (CT/TT) were significantly associated with increased risk of RCC (P = 0.023, OR = 1.21; 95% CI = 1.03-1.45). Besides, patients with variant genotypes (CT/TT) were more likely to develop large tumor (P = 0.003, OR = 1.47; 95% CI = 1.16-1.85) and advanced disease (P = 0.010, OR = 1.59; 95% CI = 1.12-2.26); and had a significantly unfavorable overall survival than those with the rs2839698 CC genotype (CT/TT vs. CC: Log-rank P = 0.026, HR = 2.25, 95%CI = 1.07-4.75). Furthermore, the CT/TT genotypes were associated with significantly increased expression of H19 in renal tissue. The luciferase reporter assays revealed the potential effect of rs2839698 variant on the binding of microRNAs to H19. Our results suggest that the H19 rs2839698 variant may be a genetic predictor of susceptibility and mortality of RCC. The risk effects and the functional impact of the variant on H19 still need further validation.

Zinc-induced protective effect for testicular ischemia-reperfusion injury by promoting antioxidation via microRNA-101-3p/Nrf2 pathway.

The present study was performed to determine the protective effect of Zinc on the rat testicular ischemia-reperfusion (I/R) injury and its mechanism. In vivo, the pathological changes and the apoptosis index were significantly relieved in the rats with Low-dose Zinc pretreatment, compared to the I/R group. After Low-dose Zinc treatment, the levels of tissue Malondialdehyde (MDA) were significantly decreased, while tissue antioxidant indices were significantly increased. Meanwhile, the level of NF-κB was significantly lower compared to I/R group, while the levels of Nrf2-dependent antioxidant enzymes were significantly higher in Low-dose Zinc+I/R group. In vitro, Low-dose Zinc markedly increased Leydig cell (TM3) cell viability, and relieved testicular oxidative damage via down-regulating ROS. A total of 22 differently expressed microRNAs were screened out using microRNA microarray in rat testicular tissue caused by I/R injury, especially showing that miR-101-3p was selected as the target miRNA. Furthermore, the levels of Nrf2 and NF-κB were apparently increased/decreased in TM3 cells treated with Hypoxic/Reoxygenation (H/R) after miR-101-3p mimics/inhibitor. In addition, H/R-induced testicular oxidative damage was recovered in TM3 administrated with miR-101-3p inhibitor and si-Nrf2. Therefore, this study provided a novel insight for investigating protective effect of Zinc on testicular I/R injury by promoting antioxidation via miR-101-3p/Nrf2.

[Outcomes of micro-TESE combined with ICSI for non-obstructive azoospermia].

OBJECTIVE: To investigate the clinicaleffects of micro-dissection of testicular sperm extraction (micro-TESE) and its combination with intracytoplasmic sperm injection (ICSI) in the treatment of non-obstructive azoospermia (NOA). METHODS: We retrospectively analyzed the clinical data on 130 NOA patients treated between December 2015 and December 2017, including36 cases of idiopathic NOA, 22 cases of idiopathic NOA with small testis, 18 cases of Klinefelter syndrome, 46 cases of surgically treated cryptorchidism and 8 cases of AZFc microdeletion.All the patients underwent micro-TESE and 29 of them with sperm received micro-TESE + ICSI. We observed the changes in the postoperative serum T level, analyzed the influences of different types of NOA and testicular pathology on the sperm retrieval rate (SRR) and the outcomes of ICSI. RESULTS: All the micro-TESE operations were successfully completed. The SRR was significantly higher in those surgically treated for cryptorchidism (60.9%) than in the other NOA groups (P<0.05), withstatistically significant differencesnot among the latter groups (P>0.05) but among differenttypes of testicular pathology (P<0.05), the highest in the hypospermatogenesis group (100%), very low in those with Sertoli-cell-only syndrome (11.8%), and the lowest in the cases of spermatogenesis arrest (0). The serum T level was remarkably decreased at 1 and 6 months after operation (P<0.01), but markedly higher at 6 than at 1 month (P<0.01). Of the 52 patients with sperm retrieved at micro-TESE, 29 of their spouses received ICSI, resulting in 17 pregnancies, 6 live births, and 6 spontaneous abortions. CONCLUSIONS: Micro-TESE is an effective method for the treatment of NOA, and its combination with ICSI can help the NOA patients obtain their genetic offspring.

Protective Effects of Ischemic Preconditioning-Mediated Homing of Endothelial Progenitor Cells on Renal Acute Ischemia and Reperfusion Injury in Male Rats.

BACKGROUND The objective of this study was to determine whether homing of endothelial progenitor cells (EPCs) induced by ischemic preconditioning (IPC) contributed to the protection of renal acute ischemia-reperfusion injury (IRI) in male rats. MATERIAL AND METHODS Forty male Sprague-Dawley rats were randomly divided into four groups, including sham-operated group, IRI-operated group, IPC-treated group and EPCs-treated group. Subsequently, serum samples were collected at 24 and 72 hours after reperfusion, respectively. In addition, histological examination was utilized to assess changes in renal structure. Moreover, immunohistochemical staining, quantitative real-time PCR and Western blotting analysis detected the expression levels of CD31, CD34, vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2). RESULTS Rats in the EPCS-treated group had significantly reduced levels of blood urea nitrogen and serum creatinine at 24 hours after operation, compared to rats that in the IRI-operated group. At 72 hours after reperfusion, renal function and morphology showed significant improvements in the EPCs-treated group. In addition, CD31+ and CD34+ cells that mostly accumulated in the renal medulla were significantly increased in IPC-treated group at 72 hours (p<0.05). Compared to the IRI-operated group, the number of EPCs in the kidneys was markedly increased at 72 hours following reperfusion in the IPC-treated group. In addition, expression levels of VEGF, Ang-1 and Ang-2 in the kidneys of the IPC-treated and EPCs-treated rats were significantly increased compared to the IRI-operated group. CONCLUSIONS These results provided evidence that IPC-mediated homing of EPCs played an important role in the protection of renal acute IRI, involving promotion of cell proliferation and angiogenesis through release of several angiogenic factors, such as VEGF, Ang-1, and Ang-2.

VEGF and Ang-1 promotes endothelial progenitor cells homing in the rat model of renal ischemia and reperfusion injury.

The aim of this study was to determine whether vascular endothelial growth factor (VEGF) and angiopoietin-1 (Ang-1) promoted the mobilization and recruitment of endothelial progenitor cells (EPCs) to protect kidneys from ischemia and reperfusion injury (IRI) in male rats. At 24 h and 72 h after reperfusion, serum samples were respectively collected for renal function. Besides, kidney tissues were harvested to observe renal morphology changes. Subsequently, VEGF, Ang-1 and angiopoietin-2 (Ang-2) expression levels in different groups were measured at the indicated time points after reperfusion. Compared with IRI-operated group, rats that were intervened with EPCs significantly reduced in the levels of blood urea nitrogen, serum creatinine at 24 hours and 72 hours, particularly in injecting EPCs suspension liquid transfected by VEGF165-adenovirus and Ang-1-adenovirus. At 72 hours after reperfusion, renal function and morphology were exhibited significant improvements in two EPCs-transfected VEGF165-adenovirus and Ang-1-adenovirus groups. In addition, expression levels of VEGF, Ang-1 and Ang-2 in the kidneys of EPCs-treated rats which were transfected by VEGF165-adenovirus and Ang-1-adenovirus were markedly increased compared to rats subjected to IRI. The present work suggested that VEGF and Ang-1 might play important roles in the protective effect of homing of EPCs on renal acute IRI.

3D shape measurement based on projection of triangular patterns of two selected frequencies.

In this paper, a temporal shift unwrapping technique is presented for solving the problem of shift wrapping associated with spatial shift estimation (SSE)-based fringe pattern profilometry (FPP). Based on this technique, a novel 3D shape measurement method is proposed, where triangular patterns of two different spatial frequencies are projected. The patterns of the higher frequency are used to implement the FPP, and the one with lower frequency is utilized to achieve shift unwrapping. The proposed method is able to solve the shift unwrapping problem associated with the existing multi-step triangular pattern FPP by projection of an additional fringe pattern. The effectiveness of the proposed method is verified by experimental results, where the same accuracy as existing multi-step triangular pattern FPP can be achieved, but enabling the measurement of objects with complex surface shape and high steps.

[Effects of endothelial progenitor cells on spermatogenic functions in testicular detorsion in rats].

OBJECTIVE: To investigate the effects of transplanted endothelial progenitor cells (EPCs) on the spermatogenic functions in testicular detorsion. METHODS: Bone-marrow-derived EPCs were obtained from rats and transfected by enhanced green fluorescent protein adenovirus (Ad-eGFP). The rats were divided into 3 groups (n = 6 each). In the sham group, left testis was not twisted. In the ischemia reperfusion injury (IRI) group, 1 ml saline was injected into the femoral vein of each rat after testicular detorsion. In the EPCs group, 1 ml EPCs suspension (1.0 × 10(6) EPCs) was injected into each rat after testicular detorsion. The Ad-eGFP transfected EPCs were injected into the 3 additional rats of testicular torsion-detorsion. At Day 5 post-transplantation, the characteristics of transplanted EPCs homing were detected. And the pathological changes and apoptotic cells/seminiferous tubules in left testis were examined. RESULTS: When the value of multiplication of infection (MOI) was at 50, the transfection rate of EPCs by Ad-eGFP exceeded 73.7%. At Day 5 post-treatment, the cells exhibiting green fluorescence were detected in left testis. The germ cells in rats of the sham group were normal. And the ratio of apoptotic cells to seminiferous tubules was 0.09 ± 0.02. The germ cells in rats of the IRI group were much fewer. And the ratio of apoptotic cells to seminiferous tubules was 2.82 ± 0.81. As compared with the IRI group, seminiferous epithelium was thicker in the EPCs group. And the ratio of apoptotic cells to seminiferous tubules was 0.32 ± 0.09 in the EPCs group. It was much smaller than that in the IRI group. There was significant difference (P < 0.01). CONCLUSION: The transplantation of EPCs is effective for treating the spermatogenic dysfunctions caused by testicular torsion so as to greatly enhance the spermatogenic functions.