Whole-transcriptome sequencing profiling identifies functional and prognostic signatures in patients with PTPRZ1-MET fusion-negative secondary glioblastoma multiforme.

Gliomas are the most common type of primary brain tumor in adults with a high mortality rate. Low-grade gliomas progress to glioblastoma multiforme (GBM) in the majority of cases, forming secondary GBM (sGBM), followed by rapid fatal clinical outcomes. Protein tyrosine phosphatase receptor type Z1 (PTPRZ1)-MET proto-oncogene receptor tyrosine kinase (MET) (ZM) fusion has been identified as a biomarker for sGBM that is involved in glioma progression, but the mechanism of gliomagenesis and pathology of ZM-negative sGBM has remained to be fully elucidated. A whole-transcriptome signature is thus required to improve the outcome prediction for patients with sGBM without ZM fusion. In the present study, whole-transcriptome sequencing on 42 sGBM samples with or without ZM fusion from the Chinese Glioma Genome Atlas database identified mRNAs with differential expression between patients with and without ZM fusion and the most significant survival-associated genes were identified. A 6-gene signature was identified as a novel prognostic model reflecting survival probability in patients with ZM-negative sGBM. Clinical characteristics in patients with a high or low risk score value were analyzed with the Kaplan-Meier method and a two-sided log-rank test. In addition, ZM-negative sGBM patients with a high risk score exhibited an increase in immune cells, NF-κB-induced pathway activation and a decrease in endothelial cells compared with those with a low risk score. The present study demonstrated the potential use of a next-generation sequencing-based cancer gene signature in patients with ZM-negative sGBM, indicating possible clinical therapeutic strategies for further treatment of such patients.

Effects of Incretin-based Therapies on Weight-related Indicators among Patients with Type 2 Diabetes: A Network Meta-analysis.

OBJECTIVE: To evaluate the effects of incretin-based therapies on body weight as the primary outcome, as well as on body mass index (BMI) and waist circumference (WC) as secondary outcomes. METHODS: Databases including Medline, Embase, the Cochrane Library, and clinicaltrials.gov (www.clinicaltrials.gov) were searched for randomized controlled trials (RCTs). Standard pairwise meta-analysis and network meta-analysis (NMA) were both carried out. The risk of bias (ROB) tool recommended by the Cochrane handbook was used to assess the quality of studies. Subgroup analysis, sensitivity analysis, meta-regression, and quality evaluation based on the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) were also performed. RESULTS: A total of 292 trials were included in this study. Compared with placebo, dipeptidyl-peptidase IV inhibitors (DPP-4Is) increased weight slightly by 0.31 kg [95% confidence interval ( CI): 0.05, 0.58] and had negligible effects on BMI and WC. Compared with placebo, glucagon-like peptide-1 receptor agonists (GLP-1 RAs) lowered weight, BMI, and WC by -1.34 kg (95% CI: -1.60, -1.09), -1.10 kg/m 2 (95% CI: -1.42, -0.78), and -1.28 cm (95% CI: -1.69, -0.86), respectively. CONCLUSION: GLP-1 RAs were more effective than DPP-4Is in lowering the three indicators. Overall, the effects of GLP-1 RAs on weight, BMI, and WC were favorable.

Prognostic value of preoperative prognostic nutritional index in stage III gastric cancer after curative resection: a retrospective cohort study.

BACKGROUND AND OBJECTIVES: Nutrition and inflammation play a crucial role in the development of cancer. The prognostic value of the prognostic nutritional index (PNI) has been confirmed in some types of human cancers. This study analyzed the prognostic significance of the preoperative PNI in patients with stage III gastric cancer after curative surgery. METHODS AND STUDY DESIGN: In this retrospective study, we enrolled 274 patients who underwent curative operation for stage III gastric cancer. The correlation between the preoperative PNI and overall survival (OS) was analyzed using Kaplan-Meier curves and multivariate Cox regression analyses. RESULTS: The patients with a high PNI had a significantly higher median OS than did those with a low PNI (46.8 months vs 24.1 months, p=0.01). In the subgroup analysis, the survival benefit of the PNI was limited to the patients with poorly differentiated gastric cancer (high PNI, 46.8 months; low PNI, 21.8 months, p=0.004) and was not observed in those with well and moderately differentiated cancer (high PNI, 30.3 months; low PNI, 26.7 months, p=0.30). In the multivariate analysis, the PNI was an independent prognostic factor for OS. CONCLUSIONS: The PNI can be used as an independent prognostic biomarker for operable advanced gastric cancer.

siRNA Screen Identifies Trafficking Host Factors that Modulate Alphavirus Infection.

Little is known about the repertoire of cellular factors involved in the replication of pathogenic alphaviruses. To uncover molecular regulators of alphavirus infection, and to identify candidate drug targets, we performed a high-content imaging-based siRNA screen. We revealed an actin-remodeling pathway involving Rac1, PIP5K1- α, and Arp3, as essential for infection by pathogenic alphaviruses. Infection causes cellular actin rearrangements into large bundles of actin filaments termed actin foci. Actin foci are generated late in infection concomitantly with alphavirus envelope (E2) expression and are dependent on the activities of Rac1 and Arp3. E2 associates with actin in alphavirus-infected cells and co-localizes with Rac1-PIP5K1-α along actin filaments in the context of actin foci. Finally, Rac1, Arp3, and actin polymerization inhibitors interfere with E2 trafficking from the trans-Golgi network to the cell surface, suggesting a plausible model in which transport of E2 to the cell surface is mediated via Rac1- and Arp3-dependent actin remodeling.

Specific Detection of Two Divergent Simian Arteriviruses Using RNAscope In Situ Hybridization.

Simian hemorrhagic fever (SHF) is an often lethal disease of Asian macaques. Simian hemorrhagic fever virus (SHFV) is one of at least three distinct simian arteriviruses that can cause SHF, but pathogenesis studies using modern methods have been scarce. Even seemingly straightforward studies, such as examining viral tissue and cell tropism in vivo, have been difficult to conduct due to the absence of standardized SHFV-specific reagents. Here we report the establishment of an in situ hybridization assay for the detection of SHFV and distantly related Kibale red colobus virus 1 (KRCV-1) RNA in cell culture. In addition, we detected SHFV RNA in formalin-fixed, paraffin-embedded tissues from an infected rhesus monkey (Macaca mulatta). The assay is easily performed and can clearly distinguish between SHFV and KRCV-1. Thus, if further developed, this assay may be useful during future studies evaluating the mechanisms by which a simian arterivirus with a restricted cell tropism can cause a lethal nonhuman primate disease similar in clinical presentation to human viral hemorrhagic fevers.

Genome Sequence of Bivens Arm Virus, a Tibrovirus Belonging to the Species Tibrogargan virus (Mononegavirales: Rhabdoviridae).

The new rhabdoviral genus Tibrovirus currently has two members, Coastal Plains virus and Tibrogargan virus. Here, we report the coding-complete genome sequence of a putative member of this genus, Bivens Arm virus. A genomic comparison reveals Bivens Arm virus to be closely related to, but distinct from, Tibrogargan virus.

Diagnostic significance of intraoperative ultrasound contrast in evaluating the resection degree of brain glioma by transmission electron microscopic examination.

BACKGROUND: Contrast-enhanced ultrasound is a dynamic and continuous modality providing real-time view of vascularization and flow distribution patterns of different organs and tumors. In order to evaluate the diagnostic significance of intraoperative contrast-enhanced ultrasound in assessing the resection degree of brain glioma by transmission electron microscopic (TEM) examination, it is important to have specific knowledge about contrast-enhanced ultrasound. Methods : Ultrasound contrast was applied in operations of 120 cases of brain glioma, to evaluate the degree of tumor resection. Biopsy tissues were obtained the suspicious residual tumors surrounding the tumor cavity. The sensitivity and specificity of the residual tumors were determined by the intraoperative ultrasound contrast according to TEM examination results. RESULTS: There were 44 cases of low-grade gliomas and 76 cases of high-grade gliomas. Three hundred and sixty biopsy tissues were obtained. The sensitivity of intraoperative ultrasound contrast in diagnosing the residual tumor was 62.2%, while the specificity degree of it was 92.8%. The consistency coefficient of the ultrasound contrast diagnosis and TEM examination results was 0.584 (Kappa = 0.584), which was between 0.4 and 0.6, therefore it was of medium consistency. Conclusions : Intraoperative ultrasound contrast was of a high sensitivity and specificity in evaluating the excision degree of tumor. The consistency of the residual tumor rate detected, respectively, by ultrasound contrast and TEM examination was of medium consistency. The application of intraoperative ultrasound contrast can improve the resection rate of brain glioma.

Simian hemorrhagic fever virus cell entry is dependent on CD163 and uses a clathrin-mediated endocytosis-like pathway.

UNLABELLED: Simian hemorrhagic fever virus (SHFV) causes a severe and almost uniformly fatal viral hemorrhagic fever in Asian macaques but is thought to be nonpathogenic for humans. To date, the SHFV life cycle is almost completely uncharacterized on the molecular level. Here, we describe the first steps of the SHFV life cycle. Our experiments indicate that SHFV enters target cells by low-pH-dependent endocytosis. Dynamin inhibitors, chlorpromazine, methyl-ß-cyclodextrin, chloroquine, and concanamycin A dramatically reduced SHFV entry efficiency, whereas the macropinocytosis inhibitors EIPA, blebbistatin, and wortmannin and the caveolin-mediated endocytosis inhibitors nystatin and filipin III had no effect. Furthermore, overexpression and knockout study and electron microscopy results indicate that SHFV entry occurs by a dynamin-dependent clathrin-mediated endocytosis-like pathway. Experiments utilizing latrunculin B, cytochalasin B, and cytochalasin D indicate that SHFV does not hijack the actin polymerization pathway. Treatment of target cells with proteases (proteinase K, papain, α-chymotrypsin, and trypsin) abrogated entry, indicating that the SHFV cell surface receptor is a protein. Phospholipases A2 and D had no effect on SHFV entry. Finally, treatment of cells with antibodies targeting CD163, a cell surface molecule identified as an entry factor for the SHFV-related porcine reproductive and respiratory syndrome virus, diminished SHFV replication, identifying CD163 as an important SHFV entry component. IMPORTANCE: Simian hemorrhagic fever virus (SHFV) causes highly lethal disease in Asian macaques resembling human illness caused by Ebola or Lassa virus. However, little is known about SHFV's ecology and molecular biology and the mechanism by which it causes disease. The results of this study shed light on how SHFV enters its target cells. Using electron microscopy and inhibitors for various cellular pathways, we demonstrate that SHFV invades cells by low-pH-dependent, actin-independent endocytosis, likely with the help of a cellular surface protein.

CD26/DPP4 cell-surface expression in bat cells correlates with bat cell susceptibility to Middle East respiratory syndrome coronavirus (MERS-CoV) infection and evolution of persistent infection.

Middle East respiratory syndrome coronavirus (MERS-CoV) is a recently isolated betacoronavirus identified as the etiologic agent of a frequently fatal disease in Western Asia, Middle East respiratory syndrome. Attempts to identify the natural reservoirs of MERS-CoV have focused in part on dromedaries. Bats are also suspected to be reservoirs based on frequent detection of other betacoronaviruses in these mammals. For this study, ten distinct cell lines derived from bats of divergent species were exposed to MERS-CoV. Plaque assays, immunofluorescence assays, and transmission electron microscopy confirmed that six bat cell lines can be productively infected. We found that the susceptibility or resistance of these bat cell lines directly correlates with the presence or absence of cell surface-expressed CD26/DPP4, the functional human receptor for MERS-CoV. Human anti-CD26/DPP4 antibodies inhibited infection of susceptible bat cells in a dose-dependent manner. Overexpression of human CD26/DPP4 receptor conferred MERS-CoV susceptibility to resistant bat cell lines. Finally, sequential passage of MERS-CoV in permissive bat cells established persistent infection with concomitant downregulation of CD26/DPP4 surface expression. Together, these results imply that bats indeed could be among the MERS-CoV host spectrum, and that cellular restriction of MERS-CoV is determined by CD26/DPP4 expression rather than by downstream restriction factors.

[Application of intraoperative contrast-enhanced ultrasound in different pathological grades of glioma].

OBJECTIVE: To explore the feasibility and value of intra-operative contrast-enhanced ultrasound in different pathological grades of glioma. METHODS: A total of 46 patients with glioma were selected to undergo the contrast-enhanced ultrasound. They were divided into 3 groups according to their pathological grades. RESULTS: All 46 tumors were hyperechoic after examinations and the borders became much more distinct. Different level of tumors had their representative ultrasonic findings. CONCLUSIONS: Intraoperative contrast-enhanced ultrasound is quite useful for determining the position and border of tumors. And it is also excellent in detecting the residual tumor tissues post-operatively. And it can improve the resection accuracy and operative precision, especially for Grade IV glioma with internal necrosis, cyst and surrounding brain edema.

Clinical characteristics and therapeutic strategies of atypical meningioma.

BACKGROUND: Atypical meningioma accounts for about 4.7% to 7.2% of all kinds of meningiomas, which is invasive with a relatively high recurrence and mortality. The objective of this study was to investigate the clinical manifestations and therapeutic strategies of atypical meningioma. METHODS: A total of 74 patients who underwent surgical treatment and pathologically confirmed for atypical meningioma in Neurosurgery Department of Beijing Tiantan Hospital from January 2003 to December 2008 were enrolled in this study. The characteristics of the tumors as well as therapeutic regimens and follow-up data were reviewed. After surgery, 56 patients underwent radiotherapy. Patients were followed up for about 3.5 years (range, 0.5 - 6.0 years), and 58 patients completed follow-up. RESULTS: Of the 58 patients who completed follow-up, good recovery was found in 30, neurological dysfunction in 15, and death in 13. Of the 58 patients, 21 had recurrent meningioma and 18 underwent a second surgery. CONCLUSIONS: Atypical meningioma is difficult to manage, with a high recurrence rate and poor survival. The extent of tumor resection and histological grade are the key determinants of outcome. Radiation therapy can be used as an adjunctive treatment after total or partial resection.

Inadequate nutritional intake and protein-energy malnutrition involved in acute and chronic viral hepatitis Chinese patients especially in cirrhosis patients.

BACKGROUND/AIMS: Early detection and treatment of protein-energy malnutrition have great clinical significance in patients with liver disease. The aim of study was to evaluate the nutritional status and determine the prevalence of malnutrition in acute and chronic viral hepatitis in Chinese patients, especially in cirrhosis patients. METHODOLOGY: A total of 315 patients were enrolled in this project, 20 patients with acute viral hepatitis, 142 patients with chronic viral hepatitis and 153 patients with posthepatitic liver cirrhosis. Recorded dietary nutrititional intakes and evaluated nutitional status by subjective global assessment, anthropometrics and biochemical data were carried out consecutively. RESULTS: According to the Chinese Dietary Recommended Nutrient Intakes based on sex and age, 73.0% +/- 29.66% of cirrhotic patients, 68.10% +/- 28.65% of chronic hepatitis patients, and 63.38% +/- 29.24% of acute hepatitis patients had deficient energy and nutrients intake. The prevalence of malnutrition was higher in the cirrhotic group as compared with the chronic or acute hepatitis groups according to the subjective global assessment, measured as triceps skinfold thickness in females, mid-upper arm muscle circumference in both male and female patients, albumin, prealbumin, hemoglobin, and serum transferrin (p < 0.05, respectively). CONCLUSIONS: This study demonstrated that Chinese patients with acute and chronic viral hepatitis and liver cirrhosis had inadequate energy and nutritional intake. Protein-energy malnutrition was more severe in cirrhotic patients, indicating a need for early enteral and perenteral treatment.

[Energy metabolism in patients with severe chronic viral hepatitis].

OBJECTIVE: To evaluate the pattern of energy metabolism of patients with chronic viral severe hepatitis. METHODS: Resting energy expenditure (REE) in 55 patients with chronic viral severe was measured with open-circuit indirect calorimetry. Their normal REE was predicted by Harris-Benedict equation (HBE). The patients were to eat the ordinary diet from the department of nutrition of the hospital. The diet was recorded for 3 days. RESULTS: The REE value of the patients with chronic viral severe hepatitis based on cirrhosis was 19.2 +/- 6.1 kcal.kg(-1).d(-1), significantly lower than the normal predicted REE (22.1 +/- 2.2 kcal.kg(-1).d(-1), P < 0.01). The REE value of the patients with chronic viral severe hepatitis based on chronic hepatitis was 20.7 +/- 6.0 kcal.kg(-1).d(-1), significantly lower than the normal predicted REE (23.1 +/- 2.3 kcal.kg(-1).d(-1), P < 0.05). The intake of carbohydrates was 21.6 +/- 7.4 kcal.kg(-1).d(-1), significantly higher than oxidation amount (7.9 +/- 7.3 kcal.kg(-1).d(-1), P < 0.01). While the intake values of fat and protein were 3.4 +/- 1.6 kcal.kg(-1).d(-1) and 4.1 +/- 2.5 kcal.kg(-1).d(-1), both lower than the needed amounts. There was no significant correlation between energy metabolism and liver function. There was a positive correlation between glucose in serum and fat oxidation (r = 0.310, P < 0.05). The serum total protein (TP) level was positively correlated with carbohydrate oxidation (r = 0.362, P < 0.01) and RQ (r = 0.348, P < 0.01); The serum cholesterol level was positively correlated with carbohydrate oxidation (r = 0.338, P < 0.05) and RQ (r = 0.354, P < 0.01). The protein oxidation was negatively correlated with serum TP (r = -0.284, P < 0.05), and serum triglyceride (TG) (r = -0.387, P < 0.05). Fat oxidation was negatively correlated with cholesterol (r = -0.328, P < 0.05). CONCLUSION: Hypometablosim is found in severe chronic hepatitis. There is no significant correlation between energy metabolism and liver function. There is certain correlation between energy metabolism and some parameters in serum, such as glucose, TP, cholesterol, and TG. It may decrease the oxidation of fat and protein to increase the oxidation of carbohydrates.

GABA, progesterone and zona pellucida activation of PLA2 and regulation by MEK-ERK1/2 during acrosomal exocytosis in guinea pig spermatozoa.

We investigated whether GABA activates phospholipase A2 (PLA2) during acrosomal exocytosis, and if the MEK-ERK1/2 pathway modulates PLA2 activation initiated by GABA, progesterone or zona pellucida (ZP). In guinea pig spermatozoa prelabelled with [14C]arachidonic acid or [14C]choline chloride, GABA stimulated a decrease in phosphatidylcholine (PC), and release of arachidonic acid and lysoPC, during exocytosis. These lipid changes are indicative of PLA2 activation and appear essential for exocytosis since inclusion of aristolochic acid (a PLA2 inhibitor) abrogated them, along with exocytosis. GABA activation of PLA2 seems to be mediated, at least in part, by diacylglycerol (DAG) and protein kinase C since inclusion of the DAG kinase inhibitor R59022 enhanced PLA2 activity and exocytosis stimulated by GABA, whereas exposure to staurosporine decreased both. GABA-, progesterone- and ZP-induced release of arachidonic acid and exocytosis were prevented by U0126 and PD98059 (MEK inhibitors). Taken together, our results suggest that PLA2 plays a fundamental role in agonist-stimulated exocytosis and that MEK-ERK1/2 are involved in PLA2 regulation during this process.

[Application of the neuro-endoscope to the intracranial aneurismal surgery].

OBJECTIVE: To study the role of neuro-endoscope in the intracranial aneurismal surgery, analyzing its benefits, and disadvantages. METHODS: Endoscopy was used as an adjunct in the microsurgical treatment of clipping aneurysms on 88 patients with 89 aneurysms, of which 82 aneurysms in 81 patients were located in the anterior circulation and 7 were located in the posterior circulation. Keyhole approach was performed on all patients, and micro-Doppler ultrasound technique was used before and after clipping. In 84 aneurysms, endoscope-assisted microneurosurgery (EAM) was used in addition to microsurgical dissection and clipping so as to observe the neck anatomic features and perforators and to verify the optimal clipping position. Endoscope-controlled microneurosurgery was used for 5 aneurysms to observe the anatomy around aneurysm, such as the posterior communicating artery and the opposite anterior communicating artery behind the internal carotid artery. RESULTS: Postoperative angiography performed on 86 patients showed satisfactory aneurysm clipping. Operative mortality was 0. There were 7 cases of postoperative complications, with an incidence rate of 7.95%, and none case was directly related to endoscopy. CONCLUSION: Improving observation of regional anatomy because of its ability in magnification, illumination, and looking around dead angle, neuro-endoscopy is very useful for complex aneurysms.