Phenylboronic acid modification-based novel dumbbell-shaped Au-Ag nanorod SERS substrates for ultrasensitive detection of SO42.

Based on the coordination principle of Lewis acids, a 4-mercaptophenylboronic acid (4-MPBA)-modified novel dumbbell-shaped Au-Ag nanorod (4-MPBA@DS Au-AgNR) substrate was developed, which could be combined with the surface-enhanced Raman scattering (SERS) technique to detect SO42- with high sensitivity and specificity. DS Au-AgNRs synthesized in this study with a dumbbell-shaped structure were verified by finite-difference time domain (FDTD) simulation to be capable of stimulating strong localized electromagnetic enhancement (EM) at nano-edge and gap, generating a large number of "hot spots" exhibiting excellent SERS performance. The 4-MPBA modified on its surface could specifically recognize SO42-, producing a change in the spectral peak at 1382 cm-1, thus realizing highly sensitive and specific sensing of SO42-. Under optimized conditions, this SERS sensor responded rapidly to SO42- within 2 minutes and demonstrated outstanding specificity. Calculation of the ratio of the characteristic peaks at 1382 and 1070 cm-1 (I1382/I1070) enabled the quantitative detection of SO42- in the range of 1 × 10-8-1 × 10-3 M, and the detection threshold was as low as 1 nM, which was superior to those of similar detection methods. Importantly, the utility and reliability of this SERS substrate for the determination of SO42- in actual samples were evaluated using ion chromatography as the gold standard, and there was no significant difference between the two protocols (P > 0.05), and the RSD was less than 6% with a satisfactory recovery rate (97.6-102.3%). Therefore, the present protocol has the advantages of simplicity and rapidity, high sensitivity, specificity, stability, and practicability in the determination of SO42- in aqueous solution, providing a reliable solution for tracing SO42- in the fields of food safety and environmental testing.

Simultaneous detection of urea and lactate in sweat based on a wearable sweat biosensor.

Urea and lactate are biomarkers in sweat that is closely associated with human health. This study introduces portable, rapid, sensitive, stable, and high-throughput wearable sweat biosensors utilizing Au-Ag nanoshuttles (Au-Ag NSs) for the simultaneous detection of sweat urea and lactate. The Au-Ag NSs arrays within the biosensor's microfluidic cavity provide a substantial surface-enhanced Raman scattering (SERS) enhancement effect. The limit of detection (LOD) for urea and lactate are 2.35 × 10-6 and 8.66 × 10-7 mol/L, respectively. This wearable sweat biosensor demonstrates high resistance to compression bending, repeatability, and stability and can be securely attached to various body parts. Real-time sweat analysis of volunteers wearing the biosensors during exercise demonstrated the method's practicality. This wearable sweat biosensor holds significant potential for monitoring sweat dynamics and serves as a valuable tool for assessing bioinformation in sweat.

The mechanism of LZ-8-mediated immune response in the mouse model of Parkinson's disease.

Parkinson's disease (PD) is associated with microscopic changes in the brain, particularly substantia nigra (SN). Ganoderma lucidum immunoregulatory protein (rLZ-8) is might confer protective effects against PD. We developed a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced murine model of PD and determined the effects of rLZ-8 on molecular and cellular components of SN and whole brain tissue. The levels of SOD, GSH-Px, p-JAK2 and p-STAT3 in the brain tissue and SN were downregulated, while IL-6, IL-1ß, and TNF-α and MDA were upregulated. These effects were significantly reversed upon treatment rLZ-8. In summary, oxidative stress and inflammatory response in PD can be alleviated using rLZ-8.

Low androgen levels induce ferroptosis of rat penile cavernous endothelial cells.

Background: Endothelial dysfunction caused by low androgen levels in penile tissue can lead to erectile dysfunction. The exact mechanism of endothelial dysfunction has not been thoroughly studied. Objective: The study sought to verify whether low androgen levels induce ferroptosis of endothelial cells in rat penile tissue. Methods: Rat penile cavernous endothelial cells (CP-R133) were divided into a no-androgen group (Dihydrotestosterone (DHT): 0 nmol/L), very low-androgen group (DHT: 0.1 nmol/L), low-androgen group (DHT: 1 nmol/L), DHT = 10 nmol/L group, DHT (0 nmol/L) + ferrostatin-1 (Fer-1) group, DHT (0.1 nmol/L) + Fer-1 group, DHT (1 nmol/L) + Fer-1 group, DHT (10 nmol/L) + Fer-1 group. Cell viability, intracellular ferrous ion (Fe2+), malondialdehyde (MDA), GSH into oxidized glutathione (GSSG), reactive oxygen species (ROS), nitric oxide (NO), transferrin receptor 1 protein (TfR1), solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4), endothelial nitric oxide synthase (eNOS), and phospho-eNOS (p-eNOS) were detected. Outcomes: Low androgen levels could induce ferroptosis of rat penile cavernous endothelial cells in vivo by upregulating the expressions of TfR1 and ACSL4 and downregulating the expressions of SLC7A11 and GPX4. Results: Cell viability, the levels of glutathione (GSH), NO, SLC7A11, GPX4, and p-eNOS/eNOS in the DHT = 0 nmol/L group were lower than those in the other groups (P < .05). The levels of Fe2+, ROS, MDA, GSSG, TfR1, and ACSL4 in the DHT = 0 nmol/L group were higher than those in the other groups (P < .05). Cell viability and the levels of GSH, NO, SLC7A11, GPX4, and p-eNOS/eNOS in the DHT = 1 nmol/L group were lower than those in the DHT (1 nmol/L) + Fer-1 group, DHT = 10 nmol/L group, and DHT (10 nmol/L) + Fer-1 group (P < .05). The levels of Fe2+, ROS, MDA, GSSG, TfR1, and ACSL4 in the DHT = 1 nmol/L group were higher than those in the DHT (1 nmol/L) + Fer-1 group, DHT = 10 nmol/L group, and DHT (10 nmol/L) + Fer-1 group (P < .05). Clinical Implications: A ferroptosis inhibitor might be a novel drug for treating erectile dysfunction caused by low androgen level. Strengths and Limitations: The results of this study need to be further confirmed in in vitro and in human studies. Meanwhile, further investigation is needed to clarify whether low androgen levels affect ferroptosis of rat penile cavernous smooth muscle and nerve cells. Conclusion: Low androgen levels can induce ferroptosis of endothelial cells in rat penile tissue. Inhibition of ferroptosis can reverse endothelial dysfunction caused by low androgen levels.

Development and validation of a nomogram model for central venous access device-related thrombosis in hospitalized children.

This study aimed to develop and validate a nomogram model of central venous access device-related thrombosis (CRT) for hospitalized children. A total of 503 consecutive cases from a hospital in Changsha City, Hunan Province were stochastically classified into the training set and internal validation set at a ratio of 7:3, and 85 consecutive cases in two hospitals in Urumqi City, Xinjiang Uygur Autonomous Region were collected as an external validation set. Univariate analysis and multivariate analysis on CRT-related risk factors of hospitalized children were conducted, a logistic regression model was employed to establish the nomogram, and the discrimination, calibration, and decision curve analysis was performed to assess the proposed nomogram model. The nomogram model involved seven independent risk factors, including blind catheterization, abnormal liver function, central line-associated bloodstream infection, infection, number of catheter lines, leukemia, and bed rest > 72 h. The discrimination results showed that the area under the receiver operating characteristic curve of the training set, internal validation set, and external validation set was 0.74, 0.71, and 0.76 respectively, and the accuracy rates of the proposed nomogram model were 79%, 72%, and 71% in the training set, internal validation set, and external validation set. The calibration results also showed that the calibration curve had great fitness for each dataset. More importantly, the decision curve suggested that the proposed nomogram model had a prominent clinical significance. CONCLUSION: The nomogram model can be used as a risk assessment tool to reduce the missed diagnosis rate and the incidence of CRT in hospitalized children. WHAT IS KNOWN: • Central venous access device-related thrombosis is generally asymptomatic for hospitalized children, causing the missed diagnosis of central venous access device-related thrombosis easily. • No risk prediction nomogram model for central venous access device-related thrombosis in hospitalized children has been established. WHAT IS NEW: • A visual and personalized nomogram model was built by seven accessible variables (blind catheterization, abnormal liver function, central line-associated bloodstream infection, infection, number of catheter lines, leukemia, and bed rest > 72 h). • The model can effectively predict the risk of central venous access device-related thrombosis for hospitalized children.

Highly efficient degradation of ofloxacin and diclofenac by composite photocatalyst aloe-emodin/PMMA.

Photocatalysis is one of the most effective methods to remove pollutants from water. Photocatalyst is the core of photocatalysis. The composite photocatalyst combines the photosensitizer with the support and uses the photosensitivity of the photosensitizer and the stability and adsorption of the support to achieve efficient and rapid degradation of pharmaceuticals in water. In this study, natural aloe-emodin with π-conjugated structure was used as photosensitizer to react with macroporous resin polymethylmethacrylate (PMMA) under mild conditions to prepare composite photocatalysts AE/PMMAs. The photocatalyst underwent photogenerated electron migration under visible light to form •O2- and holes with high oxidation activity, which could realize efficient photocatalytic degradation of ofloxacin and diclofenac sodium and showed excellent stability, recyclability and industrial feasibility. This research has developed an efficient method of composite photocatalyst and realized the application of a natural photosensitizer in pharmaceutical degradations.

Knowledge, attitude, and practice of pediatric nurses in prevention of central venous access device-related thrombosis in hospitalized children: a nationwide cross-sectional survey.

Central venous access device-related thrombosis (CRT) is a common complication in hospitalized children. The pediatric nurses' knowledge, attitude, and practice are of great significance for CRT prevention in hospitalized children. The purpose of this study was to elaborate on the level and influencing factors of pediatric nurses' knowledge, attitude, and practice on the prevention related to CRT in hospitalized children. This national cross-sectional study was carried out in China from January 16, 2021, to April 23, 2021. A multi-stage sampling method was adopted, and 1060 pediatric nurses from 21 hospitals participated in this study. The current situation and influencing factors of pediatric nurses' knowledge, attitude, and practice were investigated by descriptive statistics, approximate t test or independent-sample t test, nonparametric Kruskal-Wallis H test, one-way analysis of variance, and multiple linear regression analysis. The relationship among pediatric nurses' knowledge, attitude, and practice was explored by the Pearson correlation analysis. Among all pediatric nurses involved in this study, 25% had insufficient knowledge, 18% had negative attitudes, and 24% had poor behaviors. The main influencing factors on the knowledge, attitude, and practice included the highest education level of pediatric nurses (ß = 0.10, P = .001), whether received training related to CRT prevention (ß = 0.09-0.14, P < .01), whether CRT-related knowledge was enough for dealing with clinical work (ß = 0.18-0.21, P < .001), and the importance of hospitals/departments on children CRT prevention (ß = 0.16-0.24, P < .001). There was a positive correlation between knowledge, attitude, and practice (r = 0.24-0.77, P < .01).    Conclusion: Pediatric nurses' CRT-prevention knowledge and practice are unsatisfactory, while their preventive attitude toward CRT prevention is optimistic. This study assists the formulation of comprehensive intervention strategies for pediatric nurses in preventing CRT in hospitalized children by hospital-related organizations and nursing managers, so as to improve the participation of pediatric nurses in CRT prevention and reduce the occurrence of CRT for hospitalized children. What is Known: • As a common complication in hospitalized children, CRT can induce many potentially serious complications. • A professional nursing team is an important prerequisite for reducing CRT incidence. What is New: • The levels of pediatric nurses' knowledge and practice are not satisfactory, while pediatric nurses' preventive attitude toward CRT prevention is optimistic. • Hospital-related organizations and nursing managers should highlight the importance of CRT prevention and encourage pediatric nurses to improve their expertise and strengthen the training related to CRT prevention.

Design, synthesis and biological evaluation of quinazoline SOS1 inhibitors.

Son of sevenless 1 (SOS1) is a vital guanine nucleotide exchange factor (GEFs) that activates rat sarcoma (Ras) protein in cells. SOS1 inhibitors can effectively inhibit the expression of downstream signaling pathways by blocking the interaction between SOS1 and Ras protein. Here, we designed and synthesized a series of quinazoline-based compounds, and conducted subsequent evaluations of their biological activities. Among them, the comparable compounds I-2 (IC50 = 20 nM, against SOS1) I-5 (IC50 = 18 nM, against SOS1) and I-10 (IC50 = 8.5 nM, against SOS1) have kinase activity equivalent to BAY-293 (IC50 = 6.6 nM, against SOS1), and I-10 also has cell activity equivalent to BAY-293, providing a theoretical reference for subsequent related researches on SOS1 inhibitors.

Low awareness of venous thromboembolism among neurosurgical hospitalized patients: a cross-sectional study.

BACKGROUND: Venous thromboembolism (VTE) including Deep Venous Thrombosis (DVT) and Pulmonary Embolism (PE), is a serious cause of patient morbidity and mortality in hospitals. Neurosurgical hospitalized patients have higher rates of immobility and bed rest, thus increasing their risk of developing VTE. This highlights the need for their thromboprophylaxis regimens. Patients' awareness of VTE is essential for promoting strategies such as early ambulation and encouraging self-assessment and self-reporting of VTE signs and symptoms. This study evaluated neurosurgical hospitalized patients' awareness of VTE and explored the influencing factors to provide a theoretical basis for nursing intervention. METHODS: We selected one tertiary level hospital in Hunan Province and randomly sampled eligible patients from each five neurosurgical units. We conducted a cross-sectional survey of the hospitalized patients of neurosurgery using the self-designed and validated VTE knowledge questionnaire, and the influencing factors were analyzed using SPSS 26.0. RESULTS: A total of 386 neurosurgical hospitalized patients completed the survey. The score of VTE knowledge in neurosurgical hospitalized patients was 13.22 (SD = 11.52). 36.0% and 21.2% of participants reported they had heard of DVT and PE, respectively. 38.9% of participants were unable to correctly identify any symptoms of VTE. The most frequently identified risk factor was 'immobility or bed rest for more than three days' (50.0% of participants), and 38.1% of patients agreed that PE could cause death. 29.5% of participants were unable to identify any prophylactic measures of VTE. The results of Negative Binomial Regression showed that the influencing factors of VTE knowledge in neurosurgical hospitalized patients were education level (P < 0.004) and sources of information related to VTE, including nurses (95% CI = 2.201-4.374, P < 0.001), and family member/friend (95% CI = 2.038-4.331, P < 0.001), Internet/TV (95% CI = 1.382-2.834, P < 0.001). Other sources included patient /pamphlet/poster /professional books (95% CI = 1.492-3.350, P < 0.001). CONCLUSIONS: This study demonstrates the lack of awareness of VTE among neurosurgical hospitalized patients. More attention must be paid to carrying out training on VTE knowledge according to different characteristics of neurosurgical hospitalized patients, so as to ensure safe and high-quality patient care.

Research progress on the role of Ndfip1 (Nedd4 family interacting protein 1) in immune cells.

Nedd4 family interacting protein 1 (Ndfip1) was first mentioned in an article in 2000. Since its discovery, related studies have shown that this protein is associated with apoptosis, neuroprotection, substance transport, ubiquitination, and immune regulation. It is noteworthy that the lack of Ndfip1 can lead to death in fetal mice. Researchers generally believe that the function of Ndfip1 is closely related to individual immune capacity and have published a large number of articles. However, a comprehensive classification of the immune regulatory function of Ndfip1 is still lacking. In this review, we will overview and discuss this new perspective, focusing on the role of Ndfip1 in the proliferation, differentiation, and cell activity of CD4+ T cells, CD8+ T cells, mast cells, and eosinophils. This review provides an updated summary of Ndfip1, which will unveil novel therapeutic targets. Finally, the conclusion is that Ndfip1 mainly plays a negative regulatory role in immune cells by maintaining the stability of the immune response and limiting its overexpression.

Effect of the icariin on endothelial microparticles, endothelial progenitor cells, platelets, and erectile function in spontaneously hypertensive rats.

OBJECTIVES: To investigate the effect of icariin on endothelial microparticles, endothelial progenitor cells, platelets, and erectile function in spontaneously hypertensive rats. MATERIALS AND METHODS: Twelve 8-week-old healthy male Wistar-Kyoto rats and 12 spontaneously hypertensive rats were randomly divided into four following groups: Wistar-Kyoto control group (normal saline 1 ml/d given by gavage), Wistar-Kyoto + icariin group (icariin 10 mg/kg × d dissolved in 1 ml normal saline and given by gavage), spontaneously hypertensive rats control group (normal saline 1 ml/d given by gavage), and spontaneously hypertensive rats + icariin group (icariin 10 mg/kg × d dissolved in 1 ml normal saline and given by gavage). Four weeks later, the maximum intracavernous pressure/mean arterial pressure, platelet count, mean platelet volume, platelet distribution width, endothelial microparticles, endothelial progenitor cells, and vitronectin receptor were measured in each group. RESULTS: Under 3 or 5 V electrical stimulation, the maximum intracavernous pressure/mean arterial pressure in the spontaneously hypertensive rats + icariin group (0.23 ± 0.03, 0.38 ± 0.02) was significantly higher compared to the spontaneously hypertensive rats control group (0.12 ± 0.02, 0.20 ± 0.02) (p<0.05). Platelet count, mean platelet volume, and platelet distribution width in the spontaneously hypertensive rats + icariin group (1103.67 ± 107.70 × 109 /L, 9.08 ± 0.50 fl, 11.87 ± 0.45%) were significantly lower than those in the spontaneously hypertensive rats control group (1298.00 ± 89.54 × 109 /L, 9.72 ± 0.44 fl, 13.03 ± 0.59%) (all p < 0.05). Endothelial microparticles, endothelial progenitor cells, and vitronectin receptor in the spontaneously hypertensive rats + icariin group (1.01 ± 0.28%, 1.53 ± 0.65%, 2.13 ± 0.53%) were significantly lower than those in the spontaneously hypertensive rats control group (1.58 ± 0.19%, 2.71 ± 0.64%, 3.76 ± 0.52%) (all p < 0.05). Moreover, maximum intracavernous pressure/mean arterial pressure was strongly negatively correlated with platelet distribution width and vitronectin receptor (r > 0.7), and maximum intracavernous pressure/mean arterial pressure was moderately negatively correlated with mean platelet volume, endothelial microparticles, and endothelial progenitor cells (0.5 < r<0.7). CONCLUSION: Icariin may improve erectile function in spontaneously hypertensive rats by reducing the content of endothelial microparticles in blood and inhibiting the activation of the platelets. Endothelial microparticles, endothelial progenitor cells, and platelet activation-related (mean platelet volume, platelet distribution width, and vitronectin receptor) can be used as indicators for icariin to improve erectile function in spontaneously hypertensive rats.

Identification of a three-miRNA signature as a novel prognostic model for papillary renal cell carcinoma.

BACKGROUND: Papillary renal cell carcinoma (pRCC) accounting for near 20% of renal cell carcinoma is the second most common histological subtype. MiRNAs have been demonstrated to played significant roles on predicting prognosis of patients with tumors. An appropriate and comprehensive miRNAs analysis based on a great deal of pRCC samples from The Cancer Genome Atlas (TCGA) will provide perspective in this field. METHODS: We integrated the expression of mRNAs, miRNAs and the relevant clinical data of 321 pRCC patients recorded in the TCGA database. The survival-related differential expressed miRNAs (sDEmiRs) were estimated by COX regression analysis. The high-risk group and the low-risk group were separated by the median risk score of the risk score model (RSM) based on three screened sDEmiRs. The target genes, underlying molecular mechanisms of these sDEmiRs were explored by computational biology. The expression levels of the three sDEmiRs and their correlations with clinicopathological parameters were further validated by qPCR. RESULTS: Based on univariate COX analysis (P < 0.001), eighteen differential expressed miRNAs (DEmiRs) were remarkably related with the overall survival (OS) of pRCC patients. Three sDEmiRs with the most significant prognostic values (miR-34a-5p, miR-410-3p and miR-6720-3p) were employed to establish the RSM which was certified as an independent prognosis factor and closely correlated with OS. In the verification of clinical samples, the overexpression of miR-410-3p and miR-6720-3p were detected to be associated with the advanced T-stages, while miR-34a-5p showed the reversed results. CONCLUSION: The study developed a RSM based on the identified sDEmiRs with significant prognosis prediction values for pRCC patients. The results pave the avenue for establishing and optimizing a reliable and referable risk assessing model and provide novel insight into the researches of biomarkers and clinical treatment strategies.

Tea polyphenols protect learning and memory in sleep-deprived mice by promoting AMPA receptor internalization.

Chronic sleep loss caused lots of health problems, also including cognition impairment. Tea is one of the most popular drinks when people stay up late. Nevertheless, the effects of tea on sleep deprivation-induced cognition impairment are still unclear. In the present study, we found 24-h sleep deprivation (S-DEP) increased membrane α-amino-3-hydroxy-5-methyl-4-isoxa-zolep-propionate (AMPA) receptor level through a tumor necrosis factor α (TNFα)-dependent pathway in hippocampi. Blocking elevated TNFα level can protect S-DEP mice from impaired learning ability according to behavioral test. Tea polyphenols, major active compounds in green tea, suppressed TNFα production through downregulating TNFα converting enzyme (TACE) level. Meanwhile, tea polyphenols treatment could ameliorate recognition impairment and anxiety-like behaviors in S-DEP mice. The aforementioned results demonstrate cognition protective effects of tea polyphenols in S-DEP mice model, which provide a theoretical basis for the treatments of S-DEP-induced cognition impairment by targeting the TACE/TNFα/AMPA pathway.

[Inhibiting miR-186 expression alleviates mitochondrial damage in hypoxic human umbilical vein endothelial cells].

OBJECTIVE: To investigate the effect of miR-186 inhibition on the expression of hypoxia-inducible factor-1α (HIF-α) and mitochondrial function in hypoxic vascular endothelial cells. METHODS: Human umbilical vein endothelial cells (HUVECs) cultured in routine or hypoxic conditions for 6 h were examined for the expression of miR-186. A miR-186 inhibitor was transfected in the HUVECs, and the cells were subsequently cultured in hypoxic condition for 6 h to observe the changes in the mitochondrial structure under an electron microscope. The changes in the mRNA and protein expressions of HIF-1α in response to miR-186 interference were tested using real-time fluorescent quantitative PCR and Western blotting. RESULTS: The expression of miR-18 was mildly increased in HUVECs after hypoxic exposure for 6 h (P=0.0188). Interference of miR-186 expression obviously promoted the mRNA and protein expressions of HIF-1α in HUVECs. In hypoxic conditions, miR-186 interference significantly reduced mitochondrial damage in HUVECs as observed under electron microscope (P=0.0297). CONCLUSIONS: Inhibition of miR-186 protects vascular endothelial cells against hypoxic injuries by promoting HIF-α expression to lessen mitochondrial damage, suggesting the possibility of targeted miR-186 interference for treatment of hemorrhagic shock.

Loss of control over mild aversive events produces significant helplessness in mice.

Most of the pathophysiology of depression are still unknown because of its numerous disease states of distinct etiology and pathogenesis. Stressful rodent models have been used to test a number of hypotheses regarding the etiology of depression. The learned helplessness rodent model demonstrates that having no control at all over aversive events produces helplessness and depression, but the role of loss of control over aversive events in helplessness is still not reliably modelled or deeply investigated. A rodent model of helplessness produced by loss of control is closer to human conditions and is therefore more useful for novel mechanistic and pre-clinic studies. The present work proposed a triadic experimental design in which a Loss Of Control (LOC) group of mice was firstly exposed to escapable mild footshocks to acquire control, and then to inescapable shocks to lose control, with a yoked (L-Yoked) group receiving identical but always uncontrollable shocks. Although both the LOC and the L-Yoked groups developed helplessness, as compared with the naive control group, the helplessness exhibited in the LOC group was significantly more serious than that in the L-Yoked group. The difference in severity between the LOC and the L-Yoked groups demonstrates the effects of loss of control over aversive events, in addition to the effects of the aversive events per se. The LOC paradigm can be used to reproduce pathology of depression induced by loss of control over aversive life events, with a good constructive validity.

Integrative analysis of cancer driver genes in prostate adenocarcinoma.

Large­scale genomics studies have identified recurrently mutated genes in the ETS gene family, including fusions and copy number variations (CNVs), which are involved in the development of prostate adenocarcinoma (PRAD). However, the aetiology of PRAD remains to be fully elucidated. In the present study, 333 driver genes were identified using four computational tools: OncodriveFM, OncodriveCLUST, iCAGES and DrGaP. In addition, 32 driver pathways were identified using DrGaP. SPOP, TP53, SPTA1, AHNAK, HMCN1, ATM, FOXA1, CSMD3, LRP1B and FREM2 were the 10 most recurrently mutated genes in PRAD. ITGAL, TAGAP, SIGLEC10, RAC2 and ITGA4 were the five hub genes in the yellow module that were associated with the number of positive lymph nodes. Hierarchical clustering analysis of the 20 driver genes with the most frequent CNVs revealed three clusters of patients with PRAD. Cluster 3 tumours exhibited significantly higher numbers of positive lymph nodes, higher Gleason scores, more advanced cancer stages and poorer prognosis than cluster 1 and 2 tumours. A total of 48 genes were significantly associated with the number of positive lymph nodes, Gleason scores and pathologic stage in patients with PRAD. The identified set of cancer genes and pathways sheds light on the tumorigenesis of PRAD and creates avenues for the development of prognostic biomarkers and driver gene­targeted therapies in PRAD.

A Three-Gene Classifier Associated With MicroRNA-Mediated Regulation Predicts Prostate Cancer Recurrence After Radical Prostatectomy.

BACKGROUND AND OBJECTIVE: After radical prostatectomy (RP), prostate cancer (PCa) patients may experience biochemical recurrence (BCR) and clinical recurrence, which remains a dominant issue in PCa treatment. The purpose of this study was to identify a protein-coding gene classifier associated with microRNA (miRNA)-mediated regulation to provide a comprehensive prognostic index to predict PCa recurrence after RP. METHODS: Candidate classifiers were constructed using two machine-learning algorithms (a least absolute shrinkage and selector operation [LASSO]-based classifier and a decision tree-based classifier) based on a discovery cohort (n = 156) from The Cancer Genome Atlas (TCGA) database. After selecting the LASSO-based classifier based on the prediction accuracy, both an internal validation cohort (n = 333) and an external validation cohort (n = 100) were used to examined the classifier using survival analysis, time-dependent receiver operating characteristic (ROC) curve analysis, and univariate and multivariate Cox proportional hazards regression analyses. Functional enrichment analysis of co-expressed genes was carried out to explore the underlying moleculer mechanisms of the genes included in the classifier. RESULTS: We constructed a three-gene classifier that included FAM72B, GNE, and TRIM46, and we identified four upstream prognostic miRNAs (hsa-miR-133a-3p, hsa-miR-222-3p, hsa-miR-1301-3p, and hsa-miR-30c-2-3p). The classifier exhibited a remarkable ability (area under the curve [AUC] = 0.927) to distinguish PCa patients with high and low Gleason scores in the discovery cohort. Furthermore, it was significantly associated with clinical recurrence (p < 0.0001, log rank statistic = 20.7, AUC = 0.733) and could serve as an independent prognostic factor of recurrence-free survival (hazard ratio: 1.708, 95% CI: 1.180-2.472, p < 0.001). Additionally, it was a predictor of BCR according to BCR-free survival analysis (p = 0.0338, log rank statistic = 4.51). CONCLUSIONS: The three-gene classifier associated with miRNA-mediated regulation may serve as a novel prognostic biomarker for PCa patients after RP.

ZFAS1 functions as an oncogenic long non-coding RNA in bladder cancer.

Long non-coding RNA (lncRNA) ZFAS1 (zinc finger antisense 1) has been suggested to have an oncogenic role in the tumorigenesis of human malignant tumors. However, the expression status and biological function of ZFAS1 in bladder cancer is still unknown. Thus, the purpose of the present study is to explore the clinical value of ZFAS1 in bladder cancer patients, and the biological function of ZFAS1 in bladder cancer cell. In the present study, we found ZFAS1 expression was increased in bladder cancer tissues compared with paired adjacent normal tissues through analyzing the Cancer Genome Atlas (TCGA) database. Furthermore, we confirmed that levels of ZFAS1 expression were elevated in bladder cancer tissues and cell lines compared with normal bladder tissues and normal uroepithelium cell line, respectively. Then, we observed that the expression level of ZFAS1 was positively associated with clinical stag, muscularis invasion, lymph node metastasis, and distant metastasis in bladder cancer patients. The experiments in vitro suggested that knockdown of ZFAS1 repressed bladder cancer cell proliferation via up-regulating KLF2 and NKD2 expression, and inhibited cell migration and invasion via down-regulating ZEB1 and ZEB2 expression. In conclusion, ZFAS1 is overexpressed in bladder cancer, and functions as an oncogenic lncRNA in regulating bladder cancer cell proliferation, migration, and invasion.

[Lentiviral vectors carrying siRNA inhibit S1PR3 gene expression in the corpus cavernosum smooth muscle cells of rats with spontaneous hypertension].

OBJECTIVE: To screen lentiviral vectors carrying siRNA which can specifically down-regulate the gene expression of the sphingosine-1-phosphate receptor 3 (S1PR3) in the corpus cavernosum smooth muscle (CCSM) cells of rats with spontaneous hypertension (SHT) and investigate the influence of the vectors on the signaling pathways of ROCK1, ROCK2 and eNOS in the CCSM cells of SHT rats. METHODS: Using the S1PR3 mRNA sequence of the rat as an interfering target, we designed and synthesized three pairs of siRNA sequences (siRNA1, 2 and 3) targeting S1PR3 and one pair of negative control, and then constructed and packaged them into lentiviral vectors. We cultured the CCSM cells of SHT and Wistar-Kyoto (WKY) rats in vitro and randomly divided them into groups A (SHT untransfected control), B (SHT transfected and carrying negative control virus), C (SHT transfected and carrying siRNA1 targeting S1PR3), D (SHT transfected and carrying siRNA2 targeting S1PR3), E (SHT transfected and carrying siRNA3 targeting S1PR3), and F (WKY untransfected control). With the multiplicity of infection (MOI) = 60, we transfected the CCSM cells of the SHT rats with the lentiviral vector and then determined the expression of the green fluorescent protein (GFP) as well as the mRNA and protein expressions of S1PR3, ROCK1, ROCK2 and eNOS in the CCSM cells of the SHT and WKY rats by RT-PCR and Western blot. RESULTS: Gene sequencing proved the successful construction of the lentiviral vector. The transfection efficiency of the CCSM cells of the rats was >80% in groups B, C, D and E. Compared with group A, the mRNA and protein expressions of S1PR3, ROCK1 and ROCK2 exhibited no significant difference in group B but were remarkably decreased in groups C, D, E and F (P< 0.05), most significantly in group E, with the inhibition rates of the mRNA and protein expressions of S1PR3 of (34.2±2.9) and (77.7±4.7)%, those of ROCK1 of (33.3±1.4) and (51.1±7.3)%, and those of ROCK2 of (30.8±3.6) and (58.32±5.5)%, respectively. The mRNA and protein expressions of eNOS in group A showed no significant difference from those in groups B, C, D and E (P>0.05) but remarkably lower than those in group F (P< 0.05). Compared with group F, the mRNA and protein expressions of S1PR3, ROCK1 and ROCK2 were not significantly different from those in group E (P>0.05) but markedly increased in groups A, B, C and D (P< 0.05), while those of eNOS remarkably decreased in groups A, B, C, D and E (P< 0.05). CONCLUSIONS: The three constructed lentiviral vectors carrying siRNA targeting different loci of the S1PR3 gene could significantly inhibit the expression of S1P3 as well as RhoA/Rho kinase signaling pathways in the CCSM cells of SHT rats, and the vector carrying siRNA3 exhibited the highest inhibitory effect.

Cancer stem cell markers predict a poor prognosis in renal cell carcinoma: a meta-analysis.

BACKGROUND: Relevant markers of CSCs may serve as prognostic biomarkers of RCC. However, their actual prognostic significance remains inconclusive. Thus, a meta-analysis was performed to reevaluate the association of CSCs-relevant markers (CXCR4, CD133, CD44, CD105) expression with RCC prognosis more precisely. METHODS: PubMed and Embase were searched to look for eligible studies. The pooled hazard ratios (HR) with 95% confidence intervals (95% CI) were used to reassess the association of CSCs markers expression and RCC prognosis of overall survival (OS), cancer-specific survival (CSS), disease-free survival (DFS), and progression-free survival (PFS). RESULTS: There were 25 relevant articles, encompassing 2673 RCC patients, eligible for meta-analysis. Overall pooled analysis suggested that high CSCs markers expression predicted poor OS (HR, 2.10, 95% CI: 1.73-2.55) and DFS (HR, 3.77, 95% CI: 2.30-6.19). High CXCR4 expression predicted worse OS (HR, 2.57, 95% CI: 1.95-3.40), CSS (HR,1.97, 95% CI: 1.50-2.59), and DFS (HR, 5.82, 95% CI: 3.01-11.25). CD44 over-expression correlated with a poor OS(HR,1.58, 95% CI: 1.14-2.18), CSS (HR, 2.58, 95% CI: 1.27-5.23), and DFS (HR, 4.49, 95% CI: 2.12-9.53) in RCC patients. CD133 was an independent favorable prognostic factor for CSS (HR, 0.4, 95% CI: 0.29-0.54). CONCLUSIONS: The presence of CSCs markers correlates with poor RCC outcome. CSCs may be potentially utilized as prognostic markers to stratify RCC patients, probably representing also a novel potential therapeutic target.