[GNB2L1 gene expression and clinical value in hepatocellular carcinoma based on bioinformatics].

Objective: To analyze guanine nucleotide-binding protein subunit beta-2-like 1 (GNB2L1) expression based on bioinformatics, so as to evaluate its role and its relationship with survival rate during the occurrence and development of hepatocellular carcinoma. Methods: GEPIA, UALCAN and HPA databases were used to analyze the expression level of GNB2L1 and its relationship with HCC survival rate. Mutations in the GNB2L1 gene and their impact on survival were analyzed using the cBioPortal database. LinkedOmics database was used to analyze GNB2L1-related genes in HCC. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed simultaneously. STEING database was used to construct the GNB2L1 protein interaction network. TIMER database was used to analyze the relationship between GNB2L1 gene expression and immune infiltration in hepatocellular carcinoma. Differential expression of GNB2L1 in plasma platelets of HCC patients and healthy controls was analyzed using mRNA-based sequencing technology. Data between groups were compared using an independent-samples t-test. Results: GNB2L1 expression level was significantly increased in HCC tissues (P<0.05), and its expression was significantly correlated with body weight, classification and stage (P<0.05). The overall survival rate was higher in GNB2L1 low expression group (P<0.001). GNB2L1 and its related genes were related to biological process regulation, metabolic process, protein binding, oxidative phosphorylation, JAK-STAT signaling pathway, Ras signaling pathway and so on. GNB2L1 had interaction with RPS12, RPS11 and RPL19, and participated in multiple biological processes such as liver regeneration and positive regulation of endogenous apoptotic signaling pathway. GNB2L1 expression was significantly positively correlated with the infiltration degree of various immune cells in HCC (P<0.05). Cox regression analysis showed that GNB2L1 was an independent risk factor for lower survival rate in patients with HCC [Hazard ratio (95% confidence interval)=1.456 (1.034~2.051), P=0.031]. GNB2L1expression levels were significantly higher in platelets of HCC patients than that of healthy controls (10.40±1.36 vs. 9.58±0.51, t=2.194, P=0.037). Conclusion: GNB2L1 has high expression and close relationship to survival rate in HCC. Therefore, GNB2L1 may be a potential biomarker of HCC.

[A clinical analysis of 27 cases of simultaneous bilateral sudden sensorineural hearing loss].

Objective:To compare the clinical characteristics and therapeutic effects between simultaneous bilateral sudden sensorineural hearing loss（Si-BSSHL） and unilateral sudden sensorineural hearing loss（USSHL）. Method:The clinical data of 27 cases of Si-BSSHL were retrospectively analyzed, including the characteristics of the disease, clinical manifestations, audiological examination, hematological examination, and therapeutic effect, and compared with those of 139 cases of USSHL. Result:There was a statistically significant difference in the age（65.85±9.17 vs 56.95±13.18, P<0.01） and no significant difference in sex ratio（P>0.05） between the two groups. The proportion of Si-BSSHL patients with vertigo, ear fullness, hypertension, diabetes, coronary heart disease and stroke had no statistical difference compared with those of USSHL（P>0.05）, the proportion of patients with tinnitus in Si-BSSHL group was significantly lower than that in USSHL group（P<0.05）. The distribution of hearing loss degree and the type of audiometry in Si-BSSHL group were significantly different from those in USSHL group, PTA before treatment and the proportion of profound hearing loss in Si-BSSHL group was significantly lower than that in USSHL group（P<0.05）, descending audiometry type was more common in Si-BSSHL group compared to USSHL（P<0.01）. Fibrinogen in Si-BSSHL group was significantly higher than that in USSHL group（P<0.05）. No significant difference was found between the two groups with respect to mean platelet volume, blood lipid and folic acid（P>0.05）. The total treatment effective rate in Si-BSSHL group was 44.44%, while that in USSHL group was 41.73%, there was no significant difference between the two groups（P>0.05）. Conclusion:Si-BSSHL has a female preponderance and tends to occur in advanced age in our cohort. Compared to USSHL patients, Si-BSSHL patients have less profound hearing loss and more descending audiograms, and the proportion of patients with extremely severe deafness is relatively small. There is no significant difference in the therapeutic effect between the two groups.

[The role of ¹8F-FDG PET/CT for detecting nodal metastases in cN0 head neck cancer patients：a Meta-analysis].

Objective:The aim of this study is to evaluate the accuracy of ¹8F-fluorodeoxyglucose PET/CT(¹8F-FDG PET/CT) for the diagnosis of cervical node metastases in cN0 head neck cancer patients. Method:An electronic database search（PubMed，EMBASE，Cochrane Library，WanFang and CNKI databases）was performed. Updated quality assessment of diagnostic accuracy studies-2(QUADAS-2) was used to assess study quality. Data analyses were performed with Meta-Disc1.4. Result:Eight studies were included in the present Meta-analysis. For patient-specific data，the pooled sensitivity，specificity，diagnostic odds ratio（DOR），positive likelihood ratio（PLR），and negative likelihood（NLR）for ¹8F-FDG PET/CT were 0.61（95%CI0.52-0.69）,0.74（95%CI0.68-0.78），9.62（95%CI2.49-37.22），3.22（95%CI1.55-6.71）,and 0.42（95%CI0.24-0.37）,respectively. The area under the curve（AUC）was 0.804 1.The evaluation of heterogeneity，calculated the pooled diagnostic odds ratio，gave aQvalue of 21.26（P<0.05）and an I²of 81.2%. Conclusion:The results of meta-analysis suggested that ¹8F-FDG PET/CT did not provide better diagnostic accuracy than CT/MRI in detecting cervical node metastases in cN0 head and neck cancer. Considering the high heterogeneity of the included studies，potential value of PET/CT needs to be validated in future studies.

Reproductive biology of female striped marlin Kajikia audax in the western Pacific Ocean.

Length and mass data for 1260 (536 females, 683 males, 41 sex unknown) striped marlin Kajikia audax were collected at the fish markets of Tungkang, Singkang and Nanfangao from July 2004 to September 2010. Of these samples, 534 gonads (236 females and 298 males) ranging from 95 to 206 cm in eye-to-fork length (LEF ) and 8 to 88 kg in round mass (MR ), were collected. Chi-square tests indicated sex ratios were homogeneous among months in 2004 and 2006-2008, but not in 2005, 2009 and 2010; and there were significant differences in sex ratio by size. The overall sex ratio (RS ) differed significantly from the expected 0·5. Kajikia audax are sexually dimorphic and the proportions of females increased with size between 140 and 210 cm LEF . Reproductive activity was assessed using a gonado-somatic index (IG ), external appearance of the gonads and histological examination and results indicated that the spawning season occurred from April to August with a peak in June to July. Based on histological observations and the distribution of oocyte diameters, K. audax are multiple spawners and their oocytes develop asynchronously. The estimated length-at-50% maturity (LEF50 ) was c. 181 cm (c. 4·8 years of age) for females. The proportion of reproductively active females in the spawning season with ovaries containing postovulatory follicles (0·27) indicated that they spawned every 3·7 days on average. The hydrated oocyte method estimated mean ± S.D. batch fecundity (FB ) to be 4·4 ± 2·02 million eggs; average relative fecundity was 53·6 ± 13·9 oocytes g-1 MR ; and the average annual fecundity was 181·3 ± 48·3 million eggs. The parameters estimated in this study are key information for stock assessments of K. audax in the north-western and central Pacific and will contribute to the conservation, management and sustainable yield of this species.

High sensitivity of spontaneous spike frequency to sodium leak current in a Lymnaea pacemaker neuron.

The spontaneous rhythmic firing of action potentials in pacemaker neurons depends on the biophysical properties of voltage-gated ion channels and background leak currents. The background leak current includes a large K+ and a small Na+ component. We previously reported that a Na+ -leak current via U-type channels is required to generate spontaneous action potential firing in the identified respiratory pacemaker neuron, RPeD1, in the freshwater pond snail Lymnaea stagnalis. We further investigated the functional significance of the background Na+ current in rhythmic spiking of RPeD1 neurons. Whole-cell patch-clamp recording and computational modeling approaches were carried out in isolated RPeD1 neurons. The whole-cell current of the major ion channel components in RPeD1 neurons were characterized, and a conductance-based computational model of the rhythmic pacemaker activity was simulated with the experimental measurements. We found that the spiking rate is more sensitive to changes in the Na+ leak current as compared to the K+ leak current, suggesting a robust function of Na+ leak current in regulating spontaneous neuronal firing activity. Our study provides new insight into our current understanding of the role of Na+ leak current in intrinsic properties of pacemaker neurons.

Downregulation of miR-361-5p associates with aggressive clinicopathological features and unfavorable prognosis in non-small cell lung cancer.

OBJECTIVE: To assess the impact of miR-361-5p expression levels on non-small-cell lung cancer (NSCLC) survival. PATIENTS AND METHODS: A total of 183 patients with NSCLC who underwent surgery between October 2007 and April 2010 were included in this study. Expression levels of miR-361-5p were detected by using qRT-PCR. The association of miR-361-5p expression with clinicopathologic characteristics of NSCLC patients was analyzed. Kaplan-Meier Plotter was performed to identify the prognostic roles of miR-361-5p in NSCLC patients. Finally, multivariate analysis was performed using the Cox proportional hazard analysis. RESULTS: Results indicated that miR-361-5p was lowly expressed in NSCLC compared with adjacent non-malignant tissues (p < 0.01). And low miR-361-5p expression in NSCLC was significantly correlated with TNM stage (p = 0.000), lymph node metastasis (p = 0.001) and lymphatic invasion (p = 0.032). Kaplan-Meier analysis with the log-rank test indicated that low miR-361-5p expression had a significant impact on overall survival (p < 0.001). Furthermore, Multivariate analyses indicated that miR-361-5p represented an independent predictor for overall survival of NSCLC (p = 0.007). CONCLUSIONS: Our work revealed that miR-361-5p played critical roles in NSCLC progression and could represent a novel prognostic marker in NSCLC patients.

Trophic size-structure of sailfish Istiophorus platypterus in eastern Taiwan estimated by stable isotope analysis.

To examine trophic dynamics over different size classes, an isotopic study of sailfish Istiophorus platypterus life-history stages was carried out. Samples were collected from eastern Taiwan and the South China Sea during April 2009 and February 2012. A total of 263 samples (111-245 cm, lower jaw fork length, LLJFL ) were examined for changes in trophic structure in relation to LLJFL by using stable isotope analysis of carbon (δ(13) C) and nitrogen (δ(15) N). The δ(15) N values for I. platypterus ranged from 7·51 to 14·19‰ (mean ± s.d. = 12·06 ± 1·16‰) and the δ(13) C values ranged from -22·04 to -15·48‰ (mean ± s.d. = -17·62 ± 1·10‰). The δ(15) N values were positively dependent on LLJFL (r(2) = 0·377), whereas δ(13) C were negatively dependent on LLJFL (r(2) = 0·063). There were significantly different seasonal changes in nitrogen and carbon isotopic concentration, but no significant differences in concentrations between eastern Taiwan and the South China Sea were reported. The trophic level (TL ) of each LLJFL class was correlated, starting from 2·84 TL for size class I (LLJFL < 140 cm) and reaching 5·03 TL for size class VI (LLJFL > 221 cm). The mean ± s.d. TL was 4·43 ± 0·19 for all samples. The results reveal that I. platypterus occupies a wide range of trophic levels and different size classes occupy different trophic positions in the pelagic ecosystem.

Delayed innocent bystander cell death following hypoxia in Caenorhabditis elegans.

After hypoxia, cells may die immediately or have a protracted course, living or dying depending on an incompletely understood set of cell autonomous and nonautonomous factors. In stroke, for example, some neurons are thought to die from direct hypoxic injury by cell autonomous primary mechanisms, whereas other so called innocent bystander neurons die from factors released from the primarily injured cells. A major limitation in identifying these factors is the inability of current in vivo models to selectively target a set of cells for hypoxic injury so that the primarily injured cells and the innocent bystanders are clearly delineated. In order to develop such a model, we generated transgenic Caenorhabditis elegans strains where 2-3% of somatic cells were made selectively sensitive to hypoxia. This was accomplished by cell type-specific wild-type rescue in either pharyngeal myocytes or GABAergic neurons of a hypoxia resistance-producing translation factor mutation. Surprisingly, hypoxic targeting of these relatively small subsets of non-essential cells produced widespread innocent bystander cell injury, behavioral dysfunction and eventual organismal death. The hypoxic injury phenotypes of the myocyte or neuron sensitized strains were virtually identical. Using this model, we show that the C. elegans insulin receptor/FOXO transcription factor pathway improves survival when activated only after hypoxic injury and blocks innocent bystander death.

Preliminary selection for potential probiotic Bifidobacterium isolated from subjects of different Chinese ethnic groups and evaluation of their fermentation and storage characteristics in bovine milk.

A total of 29 strains of Bifidobacterium were isolated from 18 samples of human feces in different ethnic minority regions of China. All isolates were identified as Bifidobacterium longum (9 strains) and Bifidobacterium pseudocatenulatum (20 strains) based on 16S rRNA gene sequencing and phylogenetic analysis. These strains were preliminarily tested for their suitability to become probiotics by assessing their ability to survive adequately at low pH conditions and their tolerance of different concentrations of bile salts and simulated gastrointestinal juices. In vitro tests were sequentially used to predict the survival of these strains in the simulated conditions in the human gastrointestinal tract. These strains were first exposed to pH 2.5 for 3h, and 7 out of the 29 strains were discriminated from the others by their high survival rates. Out of these 7 strains, 4 were found to grow and survive well at an even lower pH of 2.0 and in high bile salt concentration. Apart from the gastrointestinal survival capacity, both fermentation efficiency and storage characteristics are important criteria for selecting for suitable potential probiotic strains. Therefore, the fermentation efficiency in bovine milk and the bacterial viability during the storage in the resultant fermented milk were also evaluated for these 4 selected strains. In this study, we isolated and identified 29 novel Bifidobacterium strains. Based on our initial evaluation, at least 4 of them may serve as valuable resources for further dairy probiotic strain selection.

Reproductive biology of female bigeye tuna Thunnus obesus in the western Pacific Ocean.

The reproductive biology of female bigeye tuna Thunnus obesus was assessed by examining 888 fish (ranging from 84·9 to 174·4 cm fork length, LF ) caught by Taiwanese offshore longliners in the western Pacific Ocean from November 1997 to November 1998 and November to December 1999 and 258 gonad samples from these fish. The overall sex ratio of the catch during the sampling differed significantly from 0·5, but males were predominant in sizes >140 cm LF . Reproductive activity (assessed by histology), a gonado-somatic index, and the size-frequency distributions of whole oocytes indicated that spawning occurred throughout the year and the major spawning season appeared to be from February to September. The estimated sizes at 50% maturity (LF50 ) of females was 102·85 cm (95% c.i.: 90·79-110·21 cm) and the smallest mature female was 99·7 cm LF . They are multiple spawners and oocytes develop asynchronously. The proportion of mature (0·63) and reproductively active (0·70) females with ovaries containing postovulatory follicles indicated that they spawn almost daily. Batch fecundity for 15 females with the most advanced oocytes (>730 µm) ranged from 0·84 to 8·56 million eggs (mean ± s.d. = 3·06 ± 2·09). The relationships between batch fecundity (FB , in millions of eggs) and LF (cm) and round mass (MR , kg) were FB=9·91×10-14LF6·38 (r(2) = 0·84) and FB=8·89×10-4MR2·05 (r(2) = 0·80), respectively. The parameters estimated in this study are key information for stock assessments of T. obesus in the western Pacific Ocean and will contribute to the conservation and sustainable yield of this species.

Conditional survival and cause-specific mortality after autologous hematopoietic cell transplantation for hematological malignancies.

The probability of survival is conventionally calculated from autologous hematopoietic cell transplantation (aHCT). Conditional survival takes into account the changing probability of survival with time survived, but this is not known for aHCT populations. We determined disease- and cause-specific conditional survival for 2388 patients treated with aHCT over a period of 20 years at a single institution. A total of 1054 deaths (44% of the cohort) were observed: 78% attributed to recurrent disease; 9% to subsequent malignancies and 6% to cardiopulmonary disease. Estimated probability of relative survival was 62% at 5 years and 50% at 10 years from aHCT. On the other hand, the 5-year relative survival was 70, 75, 81 and 88% after having survived 1, 2, 5 and 10 years after aHCT, respectively. The cohort was at a 13.9-fold increased risk of death compared with the general population (95% confidence interval (CI)=13.1-14.8). The risk of death approached that of the general population for 10-year survivors (standardized mortality ratio (SMR)=1.4, 95% CI=0.9-1.9), with the exception of female Hodgkin's lymphoma patients transplanted before 1995 at age 40 years (SMR=6.0, 95% CI=1.9-14.0). Among those who had survived 10 years, nonrelapse-related mortality rates exceeded relapse-related mortality rates. This study provides clinically relevant survival estimates after aHCT, and helps inform interventional strategies.

Regional variation in otolith Sr:Ca ratios of African longfinned eel Anguilla mossambica and mottled eel Anguilla marmorata: a challenge to the classic tool for reconstructing migratory histories of fishes.

Otolith Sr:Ca ratios of the African longfinned eel Anguilla mossambica and giant mottled eel Anguilla marmorata from nine freshwater sites in four rivers of South Africa were analysed to reconstruct their migratory life histories between freshwater and saltwater habitats. For A. mossambica, the Sr:Ca ratios in the otolith edge differed significantly among rivers and had large effect sizes, but did not differ among sites within a river. Otolith Sr:Ca ratios did not differ among rivers for A. marmorata. When rivers were pooled, the edge Sr:Ca ratios of A. mossambica were not significantly different from those of A. marmorata. According to the river-specific critical Sr:Ca ratio distinguishing freshwater from saltwater residence, most A. mossambica and A. marmorata had saltwater habitat experience after settlement in fresh water. This was primarily during their elver stage or early in the yellow eel stage. During the middle and late yellow eel stage, freshwater residency was preferred and only sporadic visits were made to saltwater habitats. The data also suggest that regional variations in otolith Sr:Ca ratios affect the critical Sr:Ca value and are a challenge for the reconstruction of migratory life histories that should be explicitly considered to avoid bias and uncertainty.

Separation, culture and identification of SD rat corpus cavernosal endothelial cells.

The aim of the study is to investigate the methods of separation, culture and identification of Sprague Dawley (SD) rat corpus cavernosal vascular endothelial cells (CCECs). Cavernosal tissues were isolated from male SD rats. Enzymatic digestion was applied to separate CCECs. Purified cells were obtained using immunomagnetic beads and flow cytometric cell sorting and subcultured in EMG-2 medium. The growth curve of CCECs was measured by the tetrazolium salt 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The cells were identified by von Willebrand factor (vWF) using immunofluorescence, and the positive percentage of vWF expression was detected by flow cytometry. The monomorphic cobblestone-like cells were observed by microscopy. High purification was obtained using immunomagnetic beads. After 2 days of incubation, cells entered the logarithmic growth phase and reached a plateau on the fifth day. The vWF expression in cytoplasm was positive. The purity of cells was 95.8%, which was tested by flow cytometry. SD rat CCECs can be separated and cultured successfully by the method of enzymatic digestion, immunomagnetic beads and flow cytometric cell sorting.

Health behaviors and cancer screening practices in long-term survivors of hematopoietic cell transplantation (HCT): a report from the BMT Survivor Study.

Patients undergoing hematopoietic cell transplantation (HCT) are at increased risk of chronic health conditions, including second malignant neoplasms and cardiovascular disease. Little is known about health behaviors and cancer screening practices among HCT survivors that could moderate the risk of these conditions. The BM transplant survivor study examined health behaviors and cancer screening practices in individuals who underwent HCT between 1976 and 1998, and survived 2+ years. Health behavior was deemed as high risk, if an individual was a current smoker and if they reported risky alcohol intake (≥4 drinks per day (males), ≥3 drinks per day (females)) on days of alcohol consumption. Cancer screening assessment was per American Cancer Society recommendations. There were 1040 survivors: 42.7% underwent allogeneic HCT; 43.8% were female; median time from HCT: 7.4 years (range 2.0-27.7 years). Median age at study participation: 43.8 years (range 18.3-73.0 years). Multivariate regression analysis revealed younger age (<35 years) at study participation (Odds ratio (OR)=4.7; P<0.01) and lower education (

LL-37 expression in the skin in systemic lupus erythematosus.

The objective of this study was to investigate the expression and relationship of LL-37, plasmacytoid dendritic cells (pDCs) and interferon-alpha (IFN-α) in skin in systemic lupus erythematosus (SLE), and their role in SLE pathogenesis. Skin biopsies were taken from nine SLE patients and six healthy volunteers. Expression of LL-37, pDCs and IFN-α in skin specimens and consecutive sections of skin was detected with an immunohistochemical technique (IH); the expression of LL-37 and pDCs in the samples was detected with in situ hybridization (ISH). The expression levels of LL-37, pDCs and IFN-α were significantly higher in SLE skin than in that of healthy controls (p < 0.001) with either the IH or the ISH technique, and the location of positive expression in consecutive sections was similar. Correlation analysis showed that the expression levels of LL-37, pDCs and IFN-α correlated positively with each other. In conclusion, the expression of LL-37, pDCs and IFN-α was increased in the skin of patients with active SLE. It is necessary to study further the role of LL-37 in the pathogenesis of SLE, and the exact relationship among LL-37, pDCs and IFN-α.

Late effects in survivors of acute leukemia treated with hematopoietic cell transplantation: a report from the Bone Marrow Transplant Survivor Study.

The Bone Marrow Transplant Survivor Study is a retrospective cohort study in which participants who received hematopoietic cell transplantation (HCT) between 1974 and 1998 and survived for 2 years completed a 255-item questionnaire on late effects occurring after HCT. There were 281 survivors with acute myeloid leukemia (AML) and 120 with acute lymphoblastic leukemia (ALL). Siblings of participants (n=319) were recruited for comparison. Median age at interview was 36.5 years for survivors and 44 years for siblings. Median follow-up after HCT was 8.4 years. Conditioning included total body irradiation in 86% of AML and 100% of ALL subjects. The frequencies of late effects did not differ between ALL and AML survivors. Compared with siblings, survivors had a higher frequency of diabetes, hypothyroidism, osteoporosis, exercise-induced shortness of breath, neurosensory impairments and problems with balance, tremor or weakness. In multivariable analysis, the risk of these outcomes did not differ by diagnosis. Survivors after allogeneic HCT had higher odds of diabetes (odds ratio (OR)=3.9, P=0.04), osteoporosis (OR=3.1, P=0.05), abnormal sense of touch (OR=2.6, P=0.02) and reported their overall health as fair or poor (OR=2.2, P=0.03). Ongoing surveillance for these late effects and appropriate interventions are required to improve the health status of ALL and AML survivors after HCT.