Spy1, a unique cell cycle regulator, alters viability in ALS motor neurons and cell lines in response to mutant SOD1-induced DNA damage.

Increasing evidence indicates that DNA damage and p53 activation play major roles in the pathological process of motor neuron death in amyotrophic lateral sclerosis (ALS). Human SpeedyA1 (Spy1), a member of the Speedy/Ringo family, enhances cell proliferation and promotes tumorigenesis. Further studies have demonstrated that Spy1 promotes cell survival and inhibits DNA damage-induced apoptosis. We showed that the Spy1 expression levels were substantially decreased in ALS motor neurons compared with wild-type controls both in vivo and in vitro by qRT-PCR, western blotting, and Immunoassay tests. In addition, we established that over-expression of human SOD1 mutant G93A led to a decreased expression of Spy1. Furthermore, DNA damage response was activated in SOD1G93A-transfected cells (mSOD1 cells). Moreover, decreased Spy1 expression reduced cell viability and further activated the DNA damage response in mSOD1 cells. In contrast, increased Spy1 expression improved cell viability and inhibited the DNA damage response in mSOD1 cells. These results suggest that Spy1 plays a protective role in ALS motor neurons. Importantly, these findings provide a novel direction for therapeutic options for patients with ALS as well as for trial designs, such as investigating the role of oncogenic proteins in ALS.

Downregulation of Homer1b/c in SOD1 G93A Models of ALS: A Novel Mechanism of Neuroprotective Effect of Lithium and Valproic Acid.

BACKGROUND: Mutations in the Cu/Zn superoxide dismutase (SOD1) gene have been linked to amyotrophic lateral sclerosis (ALS). However, the molecular mechanisms have not been elucidated yet. Homer family protein Homer1b/c is expressed widely in the central nervous system and plays important roles in neurological diseases. In this study, we explored whether Homer1b/c was involved in SOD1 mutation-linked ALS. RESULTS: In vitro studies showed that the SOD1 G93A mutation induced an increase of Homer1b/c expression at both the mRNA and protein levels in NSC34 cells. Knockdown of Homer1b/c expression using its short interfering RNA (siRNA) (si-Homer1) protected SOD1 G93A NSC34 cells from apoptosis. The expressions of Homer1b/c and apoptosis-related protein Bax were also suppressed, while Bcl-2 was increased by lithium and valproic acid (VPA) in SOD1 G93A NSC34 cells. In vivo, both the mRNA and protein levels of Homer1b/c were increased significantly in the lumbar spinal cord in SOD1 G93A transgenic mice compared with wild type (WT) mice. Moreover, lithium and VPA treatment suppressed the expression of Homer1b/c in SOD1 G93A mice. CONCLUSION: The suppression of SOD1 G93A mutation-induced Homer1b/c upregulation protected ALS against neuronal apoptosis, which is a novel mechanism of the neuroprotective effect of lithium and VPA. This study provides new insights into pathogenesis and treatment of ALS.

[Effects of performing pelvic lymph node dissection before versus after radical cystectomy].

OBJECTIVE: To explore the efficacies of extended pelvic lymph node dissection (e-PLND) before or after radical cystectomy (RC). METHODS: From January 2003 to January 2013, a total of 107 patients underwent e-PLND plus RC. And their relevant clinical data were reviewed. Their median age was (62 ± 10) years. The e-PLND were divided into 10 regions and 6 groups according to the anatomic sites. Forty-seven (43.9%) underwent RC after e-PLND (group A) and 60 (56.1%) had RC before e-PLND (group B). Two groups were compared for operative duration, numbers of lymph nodes removed, metastatic rates of lymph node, dissected lymph node positive rates and operative complications. The results were analyzed with Chi-square or Student's test. RESULTS: Clinicopathological characteristics were comparable for two groups (P > 0.05). The mean operative durations of e-PLND were similar in both groups ( (83 ± 27) vs (78 ± 24) min , P > 0.05). The mean operative durations of RC were significantly shorter in group A than those in group B ( (79 ± 41) vs (113 ± 44) min, P < 0.01) . The mean number of lymph nodes removed (25.5 ± 9.7 vs 29.0 ± 8.4) and the mean number of lymph nodes removed at internal iliac (5.7 ± 2.9 vs 7.2 ± 3.5) and presacral (1.3 ± 1.1 vs 2.5 ± 1.6) regions were significantly fewer in group A than those in group B (all P < 0.05). The metastatic rates of lymph node (34.0% (16/47) vs 31.7% (19/60)), dissected lymph node positive rates (9.0% (108/1197) vs 7.5% (130/1743)) and operative complications (23.4% (11/47) vs 20.0% (12/60)) were similar in both groups (all P > 0.05). CONCLUSION: RC is performed preferably after e-PLND, and internal iliac and presacral area should be dissected for additional lymph nodes after RC.

[Expression of PIM-1 in prostate cancer tissue and its relationship with PSA recurrence].

OBJECTIVE: To explore the expression of the PIM-1 protein in prostate cancer tissue and its relationship with PSA recurrence. METHODS: We used the immunohistochemical SP method to detect the expression of the PIM-1 protein in the prostate tissues of 68 cases of prostate cancer (PCa) and 37 cases of benign prostatic hyperplasia (BPH). RESULTS: The positive rate of the PIM-1 protein expression was 67.65% (46/68) in the PCa tissue, significantly higher than 40.54% (15/37) in the BPH tissue (P<0.05). Its positive rates in PCa Gleason scores 6, 7 and 8-10 were 33.33% (7/21), 77.5% (21/28) and 94.74% (18/19), respectively, with significant between-group differences (P<0.05), and those in stages I , II, III and IV of PCa were 47.62%, 53.85%, 73.33% and 94.74%, respectively. Kaplan-Meier analysis of the results of a 36-month follow-up showed the ratios of PIM-1 expression to PSA recurrence and non-recurrence were 10/22 (45.45%) and 36/46 (78.26%), respectively, with statistically significant differences (P<0.05). CONCLUSION: PIM-1 protein expression in PCa tissue is closely related to the Gleason score and clinical stage of PCa and PSA recurrence, which suggests that the PIM-1 gene plays an important role in PCa evolution and progression, and may be an indicator for the prognosis of PCa.

Promoter methylation of H-cadherin is a potential biomarker in patients with bladder transitional cell carcinoma.

OBJECTIVES: H-cadherin, functions as a tumor suppressor, is frequently silenced by promoter methylation in human cancers. The aim of this study was to evaluate the feasibility of using H-cadherin methylation in tumor tissues as a potential biomarker in patients with bladder transitional cell carcinoma (TCC). MATERIALS AND METHODS: We examined the methylation status of H-cadherin in 133 primary bladder TCC samples and 43 normal bladder epithelial tissues using methylation-specific polymerase chain reaction (MSP) and then analyzed the associations between H-cadherin methylation and clinicopathologic features as well as patients' outcome. RESULTS: H-cadherin methylation was detected in 47 (35.3%) bladder TCC samples, but not found in controls (P = 0.0000). Moreover, H-cadherin methylation was significantly associated with advanced stage (P = 0.0006), high grade (P = 0.0165), larger tumor size (P = 0.0225), tumor recurrence (P = 0.0106), and poor prognosis (P = 0.0000). In addition, multivariate analysis indicated that H-cadherin methylation is independently associated with poor outcome and had a relative risk of death of 3.832 (P = 0.0071, 95% confidence interval: 1.443-10.176). CONCLUSIONS: The results suggest that H-cadherin methylation may be used as a potential biomarker for the malignancy of bladder TCC and as an independent prognostic biomarker in patients with bladder TCC.

Prediction of proteinase cleavage sites in polyproteins of coronaviruses and its applications in analyzing SARS-CoV genomes.

Recently, we have developed a coronavirus-specific gene-finding system, ZCURVE_CoV 1.0. In this paper, the system is further improved by taking the prediction of cleavage sites of viral proteinases in polyproteins into account. The cleavage sites of the 3C-like proteinase and papain-like proteinase are highly conserved. Based on the method of traditional positional weight matrix trained by the peptides around cleavage sites, the present method also sufficiently considers the length conservation of non-structural proteins cleaved by the 3C-like proteinase and papain-like proteinase to reduce the false positive prediction rate. The improved system, ZCURVE_CoV 2.0, has been run for each of the 24 completely sequenced coronavirus genomes in GenBank. Consequently, all the non-structural proteins in the 24 genomes are accurately predicted. Compared with known annotations, the performance of the present method is satisfactory. The software ZCURVE_CoV 2.0 is freely available at http://tubic.tju.edu.cn/sars/.

ZCURVE_CoV: a new system to recognize protein coding genes in coronavirus genomes, and its applications in analyzing SARS-CoV genomes.

A new system to recognize protein coding genes in the coronavirus genomes, specially suitable for the SARS-CoV genomes, has been proposed in this paper. Compared with some existing systems, the new program package has the merits of simplicity, high accuracy, reliability, and quickness. The system ZCURVE_CoV has been run for each of the 11 newly sequenced SARS-CoV genomes. Consequently, six genomes not annotated previously have been annotated, and some problems of previous annotations in the remaining five genomes have been pointed out and discussed. In addition to the polyprotein chain ORFs 1a and 1b and the four genes coding for the major structural proteins, spike (S), small envelop (E), membrane (M), and nuleocaspid (N), respectively, ZCURVE_CoV also predicts 5-6 putative proteins in length between 39 and 274 amino acids with unknown functions. Some single nucleotide mutations within these putative coding sequences have been detected and their biological implications are discussed. A web service is provided, by which a user can obtain the annotated result immediately by pasting the SARS-CoV genome sequences into the input window on the web site (http://tubic.tju.edu.cn/sars/). The software ZCURVE_CoV can also be downloaded freely from the web address mentioned above and run in computers under the platforms of Windows or Linux.