Elevated expression of glutaminase confers glucose utilization via glutaminolysis in prostate cancer.

Cancer cells reprogram their metabolism towards aerobic glycolysis and elevated glutaminolysis, which contributes to the aggressive phenotype. Understanding how these metabolic pathways are regulated may provide critical targets for therapeutic intervention. Glutaminase (GLS1) is a key enzyme that converts glutamine to glutamate. In this study, we show the loss of GLS1 function by RNA interference or inhibitor diminished the rates of glucose utilization, growth and invasiveness of prostate cancer cells. We propose that GLS1 positively regulates glucose uptake in addition to glutaminolysis. Further, GLS1 involves the transcriptional repression of thioredoxin interacting protein (TXNIP), which is a potent negative regulator of glucose uptake and aerobic glycolysis. Most importantly, we provided direct evidence that elevated GLS1 expression was highly correlated with the tumor stage and progression in prostate cancer patients. Together, we defined a key role for GLS1 in coupling glutaminolysis of the TCA cycle with elevated glucose uptake and consequently the growth of prostate cancer cells. These data extends the role of GLS1 in regulating cell metabolism and the clinical utility of GLS1 inhibitors in the restriction of essential nutrients.

Characterization of the complete mitochondrial genome of Eopsetta grigorjewi (Herzenstein, 1890) (Chordata: Pleuronectidae) from the Yellow Sea and its phylogenetic implications.

In this study, we determined the complete mitochondrial genome (mitogenome) of Eopsetta grigorjewi Herzenstein, 1890 (Chordata: Pleuronectidae) using Sanger sequencing technology. The total length of the mitogenome sequence of E. grigorjewi is 17,269 base pairs, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, and two ribosomal RNA genes. The overall composition of the mitogenome is estimated to be 27.5% A, 25.6% T, 29.9% C, and 17.0% G. The phylogenetic relationships of 15 Pleuronectidae species were constructed based on the 13 PCGs by the maximum-likelihood approach using IQtree software.

Characterization of the complete mitochondrial genome of Laomedia astacina De Haan, 1841 (Decapoda: Laomediidae) and its phylogenetic implications.

In this study, we determined the complete mitochondrial genome (mitogenome) of Laomedia astacina De Haan, 1841 using next-generation sequencing technology. The total length of the mitogenome sequence of L. astacina is 14,795 base pairs, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, and two ribosomal RNA genes. The overall composition of the mitogenome is estimated to be 35.3% A, 38.0% T, 13.8% C, and 12.9% G, indicating that the L. astacina mitogenome is rich in A + T (73.3%). The phylogenetic relationships of 13 decapod species were constructed based on the 13 PCGs by the maximum-likelihood approach using IQtree software.

Survival after radical cystectomy for bladder cancer: Multicenter comparison between minimally invasive and open approaches.

OBJECTIVE: To investigate oncological outcomes in patients with bladder cancer who underwent minimally invasive radical cystectomy (MIRC) or open radical cystectomy (ORC). METHODS: We identified patients with bladder cancer who underwent radical cystectomy (RC) in 13 centers of the Chinese Bladder Cancer Consortium (CBCC). Perioperative outcomes were compared between MIRC and ORC. The influence of surgical approaches on overall survival (OS) and cancer-specific survival (CSS) in the entire study group and subgroups classified according to pathologic stage or lymph node (LN) status was assessed with the log-rank test. Multivariable Cox proportional hazard models were used to evaluate the association among OS, CSS and risk factors of interest. RESULTS: Of 2 098 patients who underwent RC, 1 243 patients underwent MIRC (1 087 laparoscopic RC and 156 robotic-assisted RC, respectively), while 855 patients underwent ORC. No significant differences were noted in positive surgical margin rate and 90-day postoperative mortality rate. MIRC was associated with less estimated blood loss, more LN yield, higher rate of neobladder diversion, longer operative time, and longer length of hospital stay. There was no significant difference in OS and CSS according to surgical approaches (p=0.653, and 0.816, respectively). Subgroup analysis revealed that OS and CSS were not significantly different regardless of the status of extravesical involvement or LN involvement. Multivariable Cox regression analyses showed that the surgical approach was not a significant predictor of OS and CSS. CONCLUSIONS: Our study showed that MIRC was comparable to conventional ORC in terms of OS and CSS.

Genetic effects of released swimming crab (Portunus trituberculatus) on wild populations inferred from mitochondrial control region sequences.

Along the coast of Shandong Province in China, an extensive hatchery-release programme has been conducted for more than 10 years. However, no information has been reported concerning the long-term effects on the wild population resulting from the release of large numbers of juveniles in Portunus trituberculatus. In this study, sequence variation of swimming crab P. trituberculatus based on the mitochondrial control region was investigated for 946 swimming crabs at the releasing sites from 2012 to 2014. The result showed that the wild P. trituberculatus was characterized by high genetic diversity indices, and not significantly different from the previous study. Low FST values were estimated among the groups of different years and different sites, which suggested no genetic differentiation found in the wild population after the stock enhancement programme. Hence, the long-term extensive hatchery release programme has not affected the genetic structure of wild P. trituberculatus populations along the coast of Shandong Province.

PBOV1 promotes prostate cancer proliferation by promoting G1/S transition.

Prostate cancer (PC) is one of the leading causes of cancer death in men, and thus, finding new regulators is critical for PC therapy. Prostate and breast cancer overexpressed 1 (PBOV1) is overexpressed in breast, prostate, and bladder cancers, as it is upregulated in the serum of patients with PC, but the role of PBOV1 in PC has not been studied. In this article, we found that PBOV1 was indeed overexpressed in PC cells; PBOV1 overexpression promoted cell proliferation and colony formation ability and arrested cell cycle in the G0/G1 phase and tumorigenicity ability in vitro, whereas knockdown of PBOV1 reduced these effects. Further analysis of PBOV1 overexpression inhibited cell cycle inhibitors, P21 and P27, and increased the phosphorylation level of Rb and cyclin D1 expression, suggesting that PBOV1 promoted cell proliferation through promoting G1/S transition.

Octyl-a-cyanoacrylate adhesive in the treatment of tibial transverse fracture in rabbits.

OBJECTIVE: To observe the effect of octyl-a-cyanoacrylate upon bone healing and its degradation in vitro after middle tibial transverse fracture in rabbitsì and to establish treatment of higher efficacy with the application of octyl-a-cyanoacrylate. METHODS: Middle tibial transverse fracture model of New Zealand rabbits was established. In the experimental group, internal fixation with 2 mm Kirschner wires was performed and the broken ends were fixed with octyl-a-cyanoacrylate. In the control group, only internal fixation with 2 mm Kirschner wires was conducted. Animals were killed at preset time intervals of 2, 4, 6, 8, 10 and 12 weeks postoperatively and samples were harvested. RESULTS: Two weeks after operation, clear fracture lines were observed in both the experimental and the control groups. Fibrous soft tissue connection was noted between the broken ends and there was soft tissue adhesion around the fracture site. There was no callus formation and the broken ends were surrounded by adhesive soft tissues. Obvious external callus formation was confirmed at 8 weeks after operation in both groups with partial disappearance of fracture lines. Ten and twelve weeks after the operation, fracture lines disappeared completely and there was obvious external callus formation and bone union. In the fourth week, fibrous cells and chondrocytes were found to grow into the colloid and surround it at the 6th week. The adhesive material was degraded and gradually absorbed at the 8th week. Chondrification was observed. CONCLUSIONS: Two weeks after fixation for tibial fracture in rabbits, octyl-a-cyanoacrylate begins in vivo degradation. Chondrocytes and fibrocytes gradually grow into the degradation area and surround the adhesive material, which broke into pieces at 8 weeks. Complete degradation and disappearance of the adhesive material is present between 10 and 12 weeks. No barrier effect hampering fracture healing is noted.

Flank-suspended versus prone percutaneous nephrolithotomy: changes of haemodynamics, arterial blood gases and subjective feelings.

BACKGROUND: The flank-suspended position was adopted in percutaneous nephrolithotomy (PCNL), and haemodynamics, blood gas variables and subjective feelings were examined with an attempt to explore the effect of the operative position in PCNL on the body's inner environment and patient comfort. OBJECTIVE: The influence of the flank-suspended and prone position on haemodynamics, arterial blood gases and subjective feelings in patients receiving PCNL was examined. DESIGN, SETTING AND PARTICIPANTS: A total of 100 patients with kidney stones who underwent PCNL during January 2010 to January 2011 were divided into flank-suspended groups (n = 50) and prone groups (n = 50) at random in terms of the operative position. The blood pressure, heart rate, respiratory frequency and oxyhaemoglobin saturation and blood gas variables were determined at different time points (before the operation, after position change, 30 min after the start the operation and immediately after the operation). Visual analogue scale (VAS) scoring system was employed to define the posture comfort, dyspnoea and puncture-site pain 24 h postoperation in the patients. All the measures were compared between patients in different positions at different time points. STATISTICAL ANALYSIS: Paired t-test was employed in the comparison of measures detected at different time points in the same group. Two-group comparison was subjected to t-test. A p value less than 0.05 was considered statistically significant. RESULTS AND LIMITATIONS: The blood pressure was decreased within and after the operation in both groups, substantially in the prone group, significantly lower than that before the operation (p<0.05). No significant differences in the heart rate, respiratory frequency and oxyhaemoglobin saturation were noted among the different time points in the same group. Blood gas analysis showed that pH value and base excess were profoundly reduced within and after the operation in the two groups, significantly lower than those before the operation, and the decrease was most manifest in the prone group. There was no difference in the blood sodium and potassium among the different time points in each group. The flank-suspended group was superior to the prone group with regard to posture comfort and dyspnoea degree but not puncture-site pain 24 h postoperation. CONCLUSIONS: Flank-suspended and prone PCNL affects the haemodynamics, blood gas variables and subjective feelings of patients to a varying degree. The flank-suspended PCNL possesses advantages over prone PCNL such as little influence on haemodynamics and blood gas variables, satisfactory posture comfort, less dyspnoea and easy access to vital sign observation.

Protective effects of hypovolemic hypotension preconditioning on cardiopulmonary function after myocardium ischemia/reperfusion injury.

OBJECTIVE: To identify the protective effects of hypovolemic hypotension preconditioning on cardiopulmonary function after myocardial ischemia/reperfusion injury and to explore the possible mechanism. METHODS: Twenty-four male white rabbits were randomly assigned to two groups. In the control group, ischemia/reperfusion animals(Group I/R, n=10) were subjected to thirty-minute occlusion of left anterior descending coronary artery and two-hour reperfusion. Animals in hypovolemic hypotension preconditioning group (Group HHP, n=14) experienced brief systemic ischemia preconditioning through blood withdrawal to lower blood pressure to 40%-50% of the baseline before myocardial ischemia/reperfusion. Hemodynamic parameters were recorded. Blood sample was taken to measure superoxide dismutase (SOD), malondialdehyde (MDA) and nitrogen monoxide (NO) changes with blood gas analysis. Myocardium specimens were sampled to examine apoptosis-related gene interleukin-1 beta converting enzyme (ICE) mRNA. RESULTS: Cardiac mechanical function and lung gas exchange remained stable in Group HHP with a significant increase in NO level; while in Group I/R without preconditioning, cardiopulmonary dysfunction was present after 2 h reperfusion associated with a significant reduction in NO formation and an increase in MDA (P<0.001). There was negative expression of ICE mRNA in the two groups. CONCLUSIONS: Hypovolemic hypotension preconditioning significantly improves cardiopulmonary function and increases NO formation and the protective benefit associated with hypovolemic hypotension preconditioning of the heart may be regulated through NO mediated mechanism.

Observation and establishment of an animal model of tractive spinal cord injury in rats.

OBJECTIVE: To establish an animal model of tractive spinal cord injury in rats in order to investigate its pathophysiological changes and clinical significance. METHODS: T(12)-L(3) spines were tracted longitudinally with a special spinal retractor that was put on the proccessus transverses of T(12)-L(3) vertebrae of the rat after exposing T(13)-L(2) spinal cord via dual laminectomy. At the same tine, the spinal cord function was monitored by cortical somatosensory evoked potential (CSEP). Rats were randomly divided into four groups according to the amplitude of CSEP P(1)-N(1) wave, the amount of the decreasing P(1)-N(1) wave was 30% (the 30% group), 50% (the 50% group) and 70% (the 70% group), respectively. After traction, the changes of the neural behavioral function in rats were observed and the morphological structure of the spinal cord was analyzed quantitatively with image analysis system of computer. RESULTS: With traction of spine, compared with the control group, the 30% group had no marked difference in combined behavioral score (CBS), neuron count, section area of neuron and Nissl body density, but the 50% and 70% groups had marked difference (P<0.01). Light microscope showed that the neuron volume was slightly small and the Nissl body was reduced lightly in the 30% group; the neuron space was enlarged and the neuron was degenerative, reductive, and dissolved, and the spinal cord structure was destroyed in the 50% and 70% groups. CONCLUSIONS: The animal model of tractive spinal cord injury in rats is a reproducible, graded and clinic mimic. The model in this article provides a valuable assistance in further understanding etiopathology and screening effective measures of therapy and prophylaxis of the injury.

Upstream and downstream mechanisms for the promoting effects of IGF-1 on differentiation of spermatogonia to primary spermatocytes.

AIM: The aim of this study is to delineate the mechanisms for the promoting the effects of insulin growth factor I (IGF1) on the differentiation of spermatogonia into primary spermatocytes. MAIN METHODS: We used organ culture of testicular fragments from mice to observe the effects of varying agents and siRNAs. Real-time RT-PCR and immunoblotting analysis were employed to quantify expression of genes. Luciferase reporter gene assay was employed for verifying the targeting relationship between miRNA and protein-coding genes. KEY FINDINGS: During spermatogenesis while spermatozoa pass through epididymis and vas deferens for maturation, expression of IGF1 and its receptor IGF1R, p44/ERK1 and p42/ERK2, and PI3K was all upregulated, whereas let-7 miRNA family members let-7a/b/d/e/g/ were downregulated. We established both IGF1 and IGF1R as cognate target genes for let-7; downregulation of let-7 resulted in upregulation of IGF1 and IGF1R during the early stage of differentiation from spermatogonia to primary spermatocytes. Transfection of let-7 inhibited, whereas transfection of anti-let-7 inhibitor enhanced the differentiation. The promoting effect of anti-let-7 was eliminated by PPP to block IGF1R phosphorylation. IGF1 activated ERK1/2 and PI3K and induced differentiation. PPP eliminated the activation of ERK1/2 and PI3K and inhibited the differentiation induced by IGF1. Specific inhibition of ERK1/2 by U0126 or PI3K by LY294002 reduced the IGF1-induced differentiation. Knockdown of ERK1/ERK2 or PI3K by siRNAs also blocked IGF1-induced spermatogenesis. SIGNIFICANCE: Our study therefore identified downregulation of let-7 as an upstream mechanism for IGF1/IGF1R upregulation and activation of ERK1/2 and PI3K as a downstream mechanism mediating the IGF1 signaling cascade promoting differentiation of spermatogonia to primary spermatocytes.