An Integrated Analysis of microRNAs and the Transcriptome Reveals the Molecular Mechanisms Underlying the Regulation of Leaf Development in Xinyang Maojian Green Tea (Camellia sinensis).

Xinyang Maojian (XYMJ) tea is one of the world's most popular green teas; the development of new sprouts directly affects the yield and quality of tea products, especially for XYMJ, which has hairy tips. Here, we used transcriptome and small RNA sequencing to identify mRNAs and miRNAs, respectively, involved in regulating leaf development in different plant tissues (bud, leaf, and stem). We identified a total of 381 conserved miRNAs. Given that no genomic data for XYMJ green tea are available, we compared the sequencing data for XYMJ green tea with genomic data from a closely related species (Tieguanyin) and the Camellia sinensis var. sinensis database; we identified a total of 506 and 485 novel miRNAs, respectively. We also identified 11 sequence-identical novel miRNAs in the tissues of XYMJ tea plants. Correlation analyses revealed 97 miRNA-mRNA pairs involved in leaf growth and development; the csn-miR319-2/csnTCP2 and miR159-csnMYB modules were found to be involved in leaf development in XYMJ green tea. Quantitative real-time PCR was used to validate the expression levels of the miRNAs and mRNAs. The miRNAs and target genes identified in this study might shed new light on the molecular mechanisms underlying the regulation of leaf development in tea plants.

Identification of Vitis vinifera MYB transcription factors and their response against grapevine berry inner necrosis virus.

BACKGROUND: The myeloblastosis (MYB) superfamily is the largest transcription factor family in plants that play diverse roles during stress responses. However, the biotic stress-responsive MYB transcription factors of the grapevine have not been systematically studied. In China, grapevine berries are often infected with the grapevine berry inner necrosis virus (GINV), which eventually reduces the nutritional quality and commodity value. RESULTS: The present study identified and characterized 265 VvMYB or VvMYB-related genes of the "Crimson seedless" grapevine. Based on DNA-binding domain analysis, these VvMYB proteins were classified into four subfamilies, including MYB-related, 2R-MYB, 3R-MYB, and 4R-MYB. Phylogenetic analysis divided the MYB transcription factors into 26 subgroups. Overexpression of VvMYB58 suppressed GINV abundance in the grapevine. Further qPCR indicated that among 41 randomly selected VvMYB genes, 12 were induced during GINV infection, while 28 were downregulated. These findings suggest that VvMYB genes actively regulate defense response in the grapevine. CONCLUSION: A deeper understanding of the MYB TFs engaged in GINV defense response will help devise better management strategies. The present study also provides a foundation for further research on the functions of the MYB transcription factors.

Cloning and expression analysis of SERK1 gene in Diospyros lotus.

Somatic embryogenesis receptor-like kinases (SERKs), a subfamily of receptor-like kinases, play important roles in response to abiotic stresses in addition to apomictic reproductive development in numerous plant species. The purpose of the present work was to determine if an ortholog of the SERK gene is present in the Diospyros lotus genome, isolate it and analyze its expression during embryogeny and abiotic stress. An ortholog of the SERK gene was isolated from the D. lotus genome, and designated as DlSERK1. The physical and chemical properties, protein structure, and evolutionary relationship of the DlSERK1 protein were analyzed by bioinformatics methods, and the expression of DlSERK1 gene during embryonic development and under low-temperature, salt, and drought stresses was examined through real-time quantitative PCR analysis. DlSERK1 contained 1,881 bp open reading frame encoding 626 amino acids, with a molecular mass of 69.18 kDa and pI of 5.34. DlSERK1 had strong hydrophilic property, signal peptide cleavage sites, and two transmembrane regions, indicating that DlSERK1 is a secretory protein. The secondary structure of DlSERK1 was consistent with the tertiary structure, both of which were dominated by random curls and alpha-helices. DlSERK1 had the typical structure of SERK proteins, and harbored multiple phosphorylation and glycosylation sites. Quantitative analysis showed that DlSERK1 was expressed during the embryonic development period, and the highest expression level was at 10 days post-flowering. The DlSERK1 expression level was down-regulated under low-temperature stress and up-regulated under drought and salt stresses. Our study showed that DlSERK1 was expressed in embryo development and could respond to low-temperature, drought, and salt stresses, which lays a foundation for further research on the function of SERK1 in the apomixis growth and development of environmental adaptation in D. lotus.

The antioxidant system response to drought-stressed Diospyros lotus treated with exogenous melatonin.

Drought is one of the major abiotic stresses adversely impacting the growth of persimmon, which is a widely cultivated traditional fruit tree in North China. Melatonin is a bio-stimulator involved in mediating plant responses to drought. The role of exogenous melatonin application in the drought tolerance of Diospyros lotus was examined under drought stress with different doses of melatonin (0, 10, 50, and 100 µM). Exogenous melatonin application significantly mitigated the adverse effects of drought stress on chlorophyll fluorescence, lipid peroxidation, reactive oxygen species (ROS) accumulation and nitric oxide (NO) content. The 100-µM melatonin application produced the most beneficial impacts against drought stress. The melatonin-enhanced tolerance could be attributed to improved antioxidant enzymes, reduced drought-induced ROS accumulation, and lipid peroxidation. Melatonin application activated major antioxidant enzymes such as superoxide dismutase, catalase, peroxidase, glutathione reductase, and ascorbate peroxidase. Interestingly, NO concentration was significantly higher in 10 and 50 µM melatonin treatments and lower in 100 µM melatonin treatment compared to the control. Moreover, exogenous melatonin application affected the mRNA transcript levels of several genes involved in ROS metabolism, including DlRBOHA, DlSOD, DlCAT, and DlPOD. Hence, the responses of Diospyros lotus to drought varied with different doses of melatonin. Our results provide a concrete insight into the effects of melatonin with varying doses in alleviating drought as well as a platform for its potential application in the related fields.

Microvesicles derived from human umbilical cord mesenchyme promote M2 macrophage polarization and ameliorate renal fibrosis following partial nephrectomy via hepatocyte growth factor.

The intraoperative ischemia in partial nephrectomy (PN) often leads to postoperative renal function impairment and fibrosis, which can be regulated by macrophage polarization. We have previously demonstrated that microvesicles derived from human Wharton's Jelly mesenchymal stromal cells (hWJMSC-MVs) attenuated renal ischemia-induced renal fibrosis and contained a substantial quantity of hepatocyte growth factor (HGF). Herein, we investigated whether MSC-MVs regulate macrophage polarization and ameliorate renal fibrosis following ischemia-PN via transferring HGF. A rat model of ischemia-PN was established by 45 min of left renal ischemia followed by removal of 1/3 upper left kidney. MSC-MVs were injected through the tail vein immediately after ischemia. Renal injury biomarkers were measured and histologic analysis was performed to analyze renal injury. A co-culture model of THP-1 macrophages and MSC-MVs was utilized. The expression of M1 markers and M2 markers were determined to evaluate macrophage polarization. MSC-MV administration significantly ameliorated renal inflammation, lesions, and fibrosis in ischemia-PN rats, and promoted M2 macrophage polarization both in rat remnant renal tissues and LPS-treated THP-1 cells. These effects of MSC-MVs were compromised when HGF expression was downregulated in MSC-MVs. Collectively, MSC-MVs promote M2 macrophage polarization and attenuate renal fibrosis following ischemia-PN via transferring HGF.

Microvesicles derived from human umbilical cord mesenchymal stem cells ameliorate renal ischemia-reperfusion injury via delivery of miR-21.

Microvesicles (MVs) derived from human umbilical cord mesenchymal stem cells (hUC-MSCs-MVs) and miR-21 were demonstrated to ameliorate renal ischemia-reperfusion injury (IRI). Since hUC-MSC-MVs contained a substantial quantity of miR-21, we speculated that miR-21 might account for a part of the therapeutic effects of hUC-MSCs-MVs. The human tubule epithelial (HK-2) cells were cultured under low oxygen (LO) condition to mimic a cellular IRI model. A rat model of unilateral renal IRI was established. A co-culture model of HK-2 cells and MSC-MVs was utilized to examine the therapeutic role of MSC-MVs in HK-2 cell apoptosis and mechanism. The results showed that hUC-MSCs-MVs inhibited LO-induced HK-2 cell apoptosis through transferring miR-21 to HK-2 cells. Mechanistically, miR-21 directly targeted and negatively regulated programmed cell death protein 4 (PDCD4) in HK-2 cells. Moreover, PDCD4 overexpression in HK-2 cells abrogated the hUC-MSCs-MVs-inhibited HK-2 cell apoptosis under LO condition. Additionally, the beneficial effect of MSC-MVs on rat renal IRI was partly eliminated when miR-21 was knocked down in MSCs. Taken together, MSC-MVs inhibit tubular epithelial cell apoptosis and ameliorate renal IRI, at least partially, via delivery of miR-21.

Preoperative Plasma Fibrinogen and D-dimer as Prognostic Biomarkers for Non-Muscle-Invasive Bladder Cancer.

PURPOSE: To assess the ability of preoperative plasma fibrinogen and D-dimer as biomarkers to predict survival outcomes in patients with non-muscle-invasive bladder cancer (NMIBC). PATIENTS AND METHODS: A total of 206 NMIBC patients receiving transurethral resection of bladder tumor (TURBT) were assessed in our retrospective study. The cutoff values of fibrinogen and D-dimer were determined using receiver operating characteristic curve analysis. Cox regression analyses were adopted to assess the influence of these two parameters on recurrence-free survival (RFS) and progression-free survival (PFS). RESULTS: The cutoff values of fibrinogen and D-dimer were 3.56 g/L and 0.48 µg/mL, respectively. Kaplan-Meier analysis demonstrated that high fibrinogen and D-dimer levels were significantly related to poor RFS (all P < .001) and PFS (all P < .001). Moreover, patients with elevated fibrinogen levels tended to have high tumor grade (P = .033), advanced pathologic T stage (P < .001), and multiple tumor lesions (P = .019). Significant associations of high D-dimer levels with advanced pathologic T stage (P = .026), large tumor size (P = .012), and multiple tumor lesions (P = .006) were found. In addition, multivariate analysis revealed that plasma fibrinogen and D-dimer were all independent predictive factors for RFS (P = .029 and .001, respectively) and PFS (P = .023 and .003, respectively). CONCLUSION: High levels of preoperative plasma fibrinogen and D-dimer may indicate advanced clinicopathologic features and worse prognosis, suggesting that these two coagulation parameters could be used as prognostic biomarkers for NMIBC patients.

Lipomatous hemangiopericytoma of the corpus spongiosum: a case report and review of the literature.

BACKGROUND: Hemangiopericytoma (HPC) is an uncommon soft tissue tumor arising from pericytes. The urogenital system is rarely affected. METHODS: The review of the literature used the PubMed database which was searched up to March 2015. RESULTS: Herein, we report the first case of lipomatous HPC of the corpus spongiosum in a 37-year-old man in China. The lesion presented as a quickly growing mass. Contrast enhanced CT showed a heterogeneous fatty mass with a multifocal enhancing soft-tissue component. Microscopically, the neoplasm was composed of spindle cells, a mature fat component and collagenous stroma. The mitotic index was low at 1 to 3 mitoses per 10 high-power fields. Immunohistochemically, STAT6, Bcl-2, CD99 and CDK4 were positive; CD34 and SMA were negative. The mature adipocytes were positive for S-100. Ki-67 expression was found in approximately 5% of the tumor cells. Surprisingly, there was a diffuse and strong nuclear expression of MDM2, but, no amplification of MDM2 was demonstrated by FISH. An adequate excision of the tumor was performed. CONCLUSION: No local recurrence or distant metastases occurred during the 18-month follow-up. In view of the unpredictable biological behavior of this tumor, a long follow-up period is mandatory.

Collecting duct carcinoma of the kidney: a clinicopathological study of five cases.

OBJECTIVE: To investigate the clinicopathological features of collecting duct carcinoma (CDC) and improve its diagnosis and treatment. METHODS: A retrospective analysis was performed with clinical data including follow-up results of five patients with CDC. RESULTS: A total of 5 cases, including 4 males and 1 female, were included in this analysis with the average age 54 years (range 42 to 65). Patients mainly suffered from lumbar pain, hematuria, abdominal mass and low grade fever. Four patients underwent radical nephrectomy while another received palliative nephrectomy. Lymph node metastasis occurred in 3 cases and renal hilum fat metastasis happened to 2 other cases. Tumors was located in the renal medulla and presented invasive growth. They had a tubulopapillary architecture with the hobnail-shaped cells protruding into the glandular lumen, and were accompanied by interstitial fibrosis and dysplasia of epithelial cells in collecting ducts adjacent to the tumors. One tumor was staged at AJCC II, two at AJCC III and two at AJCC IV. Postoperative interferon immunotherapy was applied in 2 cases. Patients were followed up for 5 to 18 months and the average survival time was 10 months. CONCLUSION: The CDC exhibits special clinicopathological features, high degree of malignancy and poor prognosis. The diagnosis depends on the histopathological examination. Early detection and early surgical treatment are still the main methods to improve the prognosis of patients with CDC.

Meta-analysis of gene expression data identifies causal genes for prostate cancer.

Prostate cancer is a leading cause of death in male populations across the globe. With the advent of gene expression arrays, many microarray studies have been conducted in prostate cancer, but the results have varied across different studies. To better understand the genetic and biologic mechanisms of prostate cancer, we conducted a meta-analysis of two studies on prostate cancer. Eight key genes were identified to be differentially expressed with progression. After gene co-expression analysis based on data from the GEO database, we obtained a co- expressed gene list which included 725 genes. Gene Ontology analysis revealed that these genes are involved in actin filament-based processes, locomotion and cell morphogenesis. Further analysis of the gene list should provide important clues for developing new prognostic markers and therapeutic targets.

[Urethroplasty with circular fasciocutaneous flap in the treatment of complex anterior urethral strictures].

OBJECTIVE: To summarize the experience of urethral reconstruction using circular fasciocutaneous flap for the treatment of complex anterior urethral strictures. METHOD: The circular fasciocutaneous flap was harvested from the distal penile shaft or foreskin. Then the circular configuration was converted into longitudinal strip of skin which was rotated to ventral side to repair the exposured urethral strictures using the ventral onlay method. The surveillance protocol after urethroplasty was urination observation, regularly uroflowmetry and urethrography examination. RESULTS: From Nov. 2006 to Oct. 2012, 15 cases were treated. The mean age was 45 years (20-76 years) and mean follow-up period was 18 months(3 months-3 years). Stricture was caused by chronic urethritis in 4 cases, long-term urethral catheterization in 3 cases, transurethral perfusion chem other aphy in 3 cases, transurethral prostatectomy in 3 cases and hypospadias after surgery in 2 patients. The mean stricture length was 7.0 cm (3.5-12.0 cm). The overall success rate was 80.0% (12/15). Recurrence stenosis was noted in 2 cases and diverticulum formation in 1 case. CONCLUSIONS: The penile circular fasciocutaneous flap can be used for anterior urethral stricture in nearly any length. The flap has the characteristics of hairless, adequate mobile and length, well-vascularized pedicle and easy to harvest. The onlay reconstruction provides excellent cosmetic results, less trauma, higher success rate. Therefore it should be one of the preferred techniques for complex anterior urethral stricture repair.

[High-frequency color Doppler ultrasonography: valuable for diagnosis of polyorchidism].

OBJECTIVE: To investigate the application value of high-frequency color Doppler ultrasonography in the diagnosis of polyorchidism. METHODS: We retrospectively studied 6 cases of polyorchidism diagnosed by ultrasound, 5 of which were pathologically confirmed, and analyzed the sonographic and clinical findings. RESULTS: All the 6 cases were triorchidism, 4 located in the scrotum, and 2 in the same groin with indirect hernia. The supernumerary testis was linked to the epididymis in 3 cases, 1 accompanied with repeated epididymis and vas deferens, and 2 connected to no vas deferens and epididymis. Color Doppler ultrasonography revealed blood flow signals in 4 cases, but not in the other 2. CONCLUSION: Polyorchidism has typical sonographic appearances, and high-frequency color Doppler ultrasonography plays an important role in its diagnosis.