WSSV early protein WSSV004 enhances viral replication by suppressing LDH activity.

White spot syndrome virus (WSSV) is known to upregulate glycolysis to supply biomolecules and energy for the virus's replication. At the viral genome replication stage, lactate dehydrogenase (LDH), a glycolytic enzyme, shows increased activity without any increase in expression. In the present study, yeast 2-hybrid screening was used to identify WSSV proteins that interacted with LvLDH isoform 1 and 2, and these included the WSSV early protein WSSV004. The interaction between WSSV004 and LvLDH1/2 was confirmed by co-immunoprecipitation. Immunofluorescence showed that WSSV004 co-localized with LvLDH1/2 in the cytoplasm. dsRNA silencing experiments showed that WSSV004 was crucial for WSSV replication. However, although WSSV004 silencing led to the suppression of total LvLDH gene expression during the viral late stage, there was nevertheless a significant increase in LvLDH activity at this time. We also used affinity purification-mass spectrometry to identify cellular proteins that interact with WSSV004, and found a total of 108 host proteins and 3 WSSV proteins with which it potentially interacts. Bioinformatics analysis revealed that WSSV004 and its interacting proteins might be responsible for various biological pathways during infection, including vesicular transport machinery and RNA-related functions. Collectively, our study suggests that WSSV004 serves as a multifunctional modulator to facilitate WSSV replication.

A combined bioinformatics and experimental approach identifies RMI2 as a Wnt/ß-catenin signaling target gene related to hepatocellular carcinoma.

BACKGROUND: The Wnt/ß-catenin signaling pathway plays an important role in embryogenesis and tumorigenesis. In human cancer, abnormal activity of Wnt/ß-catenin signaling pathway induces overexpressed of downstream genes, and initiate oncogene. There are several target genes known to be key players in tumorigenesis, such as c-myc, cyclin D1, MMPs or survivin. Therefore, identifying the target genes of Wnt/ß-catenin signaling pathway is important to understanding Wnt/ß-catenin-mediated carcinogenesis. In this study, we developed a combined bioinformatics and experimental approach to find potential target genes. METHODS: Luciferase reporter assay was used to analyze the promoter activity of RMI2. WST1 cell proliferation assays and transwell assays were performed to determine the proliferation and migration capacities of RMI2 overexpressing or knockdown stable hepatic cells. Finally, xenograft experiments were performed to measure the tumor formation capacity in vivo. RESULTS: The results showed that RMI2 mRNA was upregulated after LiCl treatment and Wnt3a-conditioned medium in a culture of SK-hep-1 cell lines. A chromatin immunoprecipitation (ChIP) assay showed that the ß-catenin/T cell-specific factor (TCF) complex binds to the putative TCF binding site of the RMI2 promoter. We then found a TCF binding site at - 333/- 326 of the RMI2 promoter, which is crucial for ß-catenin responsiveness in liver cell lines. RMI2 was overexpressed in hepatoma tissue and cell lines, and it promoted the migration and invasion of HCC cells. Moreover, RMI2 upregulated the expression of epithelial-mesenchymal transition (EMT) markers and the Wnt3a/ß-catenin-related genes, but silencing RMI2 had the opposite effects. Notably, the expression of RMI2 was positively correlated with the clinical data of HCC patients who had significantly shorter overall survival (OS) and disease-free survival (DFS) (Both: P < 0.05). In addition, a total of 373 HCC patients' data from the Caner Genome Atlas project (TCGA) were used to validate our findings. CONCLUSIONS: Taking all these findings together, we determined that RMI2 was a new target gene of the Wnt/ß-catenin signaling pathway. We also found that RMI2 promotes EMT markers, HCC cell invasion, and metastasis, which indicated that RMI2 is a potential target for preventing or at least mitigating the progression of HCC.

White spot syndrome virus (WSSV) modulates lipid metabolism in white shrimp.

In addition to the Warburg effect, which increases the availability of energy and biosynthetic building blocks in WSSV-infected shrimp, WSSV also induces both lipolysis at the viral genome replication stage (12 hpi) to provide material and energy for the virus replication, and lipogenesis at the viral late stage (24 hpi) to complete virus morphogenesis by supplying particular species of long-chain fatty acids (LCFAs). Here, we further show that WSSV causes a reduction in lipid droplets (LDs) in hemocytes at the viral genome replication stage, and an increase in LDs in the nuclei of WSSV-infected hemocytes at the viral late stage. In the hepatopancreas, lipolysis is triggered by WSSV infection, and this leads to fatty acids being released into the hemolymph. ß-oxidation inhibition experiment reveals that the fatty acids generated by WSSV-induced lipolysis can be diverted into ß-oxidation for energy production. At the viral late stage, WSSV infection leads to lipogenesis in both the stomach and hepatopancreas, suggesting that fatty acids are in high demand at this stage for virion morphogenesis. Our results demonstrate that WSSV modulates lipid metabolism specifically at different stages to facilitate its replication.

Effect of thermocycling-induced stress on properties of orthodontic NiTi wires.

Background/purpose: In orthodontic applications, NiTi wires are under continuous bending stress and exposed to fluctuations in temperature over long durations. The sensitivity of NiTi to temperature can have a considerable influence on its mechanical properties. This study investigated the effects of deflected NiTi wire, presented in stress-induced (detwinned) martensite microstructure, combined with thermal cycle on the microstructure and mechanical properties. Materials and methods: We tested four types of as-received orthodontic NiTi: (1) Nitinol Classic (3 M Unitek), (2) Sentalloy (Tomy), (3) 27 °C CuNiTi (Ormco) and (4) 40 °C CuNiTi (Ormco). Each group of specimens was subjected to three different testing conditions: (1) temperature fluctuations (5000 cycles) between 5 and 55 °C, (2) continuous three-point bending force and (3) combination of thermal cycling and bending stress. Results: The specimens that underwent thermocycling as well as loading exhibited a substantial narrowing in stress hysteresis, which may be attributed to crystallinity lower than that of as-received NiTi wires. Reduced crystallinity can manifest in a number of imperfections, such as dislocations and internal stress, as well as a less-organized structure. Micro X-ray diffraction (XRD) analysis revealed the existence of martensite phase in Sentalloy wires subject to thermal and stress conditions. Under loading conditions, stress-induced martensite of NiTi wires exposed to temperature fluctuations of 5-55 °C also induced cyclic changes in bending stress. In a simulated intra-oral environment, the stability of austenite↔martensite transformation decreased. Conclusion: This study determined that bending stress in conjunction with repeated temperature fluctuations can greatly affect the microstructure and mechanical properties of NiTi wires.

Antipsychotic prescription patterns and associated factors among the elderly with psychiatric illnesses.

Prescribing rate of antipsychotics in elderly patients with psychiatric illnesses has been increasing all over the world. However, there is a lack of research examining the use of antipsychotics at psychiatric hospitals. We aim to find out long-term trends in antipsychotic prescriptions and factors associated with the use of antipsychotics in the elderly population. All outpatient visits with patients aged over 65 years between 2006 and 2015 in a psychiatric hospital were included in the analysis. Demographic and clinical data, including patient age, sex, National Health Insurance status, psychiatric diagnosis and antipsychotic prescription, were retrieved through the electronic medical information system. In this study, we found that prescribing rate of antipsychotics has increased around 5.07% through the study period (from 57.25% in 2006 to 60.15% in 2015, P < 0.001). Among all antipsychotics, there was an increase in the use of second-generation antipsychotics with a simultaneous decline in the use of first-generation antipsychotics. Logistic regression analysis showed advanced age, female gender and certification for catastrophic illnesses were positively associated with the use of antipsychotics. Furthermore, patients most likely to be prescribed an antipsychotic were those with psychotic disorders, followed by dementia, and then affective or neurotic disorders. The results of this study were consistent with the worldwide trend of increasing prescriptions of second-generation antipsychotics among elderly patients with psychiatric illnesses. Despite the potential benefits of these medications on certain psychiatric symptoms, clinicians need to exercise due caution as they may also cause potentially serious side effects among the elderly population.

Progesterone receptor membrane component 1 as a potential prognostic biomarker for hepatocellular carcinoma.

AIM: To investigate the clinicopathological significance of progesterone receptor membrane component 1 (PGRMC1) and PGRMC2 in hepatocellular carcinoma (HCC). METHODS: We performed immunohistochemical staining to evaluate the estrogen receptor (ER), progesterone receptor (PR), PGRMC1, and PGRMC2 in a clinical cohort consisting of 89 paired HCC and non-tumor liver samples. We also analyzed HCC data (n = 373) from The Cancer Genome Atlas (TCGA). We correlated the expression status of PGRMC1 and PGRMC2 with clinicopathological indicators and the clinical outcomes of the HCC patients. We knocked down or overexpressed PGRMC1 in HCC cell lines to evaluate its biological significance in HCC cell proliferation, differentiation, migration, and invasion. RESULTS: We found that few HCC cases expressed ER (5.6%) and PR (4.5%). In contrast, most HCC cases expressed PGRMC1 (89.9%) and PGRMC2 (100%). PGRMC1 and PGRMC2 exhibited significantly lower expression in tumor tissue than in non-tumor tissue (P < 0.001). Lower PGRMC1 expression in HCC was significantly associated with higher serum alpha-fetoprotein expression (P = 0.004), poorer tumor differentiation (P = 0.045) and liver capsule penetration (P = 0.038). Low PGRMC1 expression was an independent predictor for worse disease-free survival (P = 0.002, HR = 2.384, CI: 1.377-4.128) in our cases, as well as in the TCGA cohort (P < 0.001, HR = 2.857, CI: 1.781-4.584). The expression of PGRMC2 did not relate to patient outcome. PGRMC1 knockdown promoted a poorly differentiated phenotype and proliferation of HCC cells in vitro, while PGRMC1 overexpression caused the opposite effects. CONCLUSION: PGRMC1 is a non-classical hormonal receptor that negatively regulates hepatocarcinogenesis. PGRMC1 down-regulation is associated with progression of HCC and is a poor prognostic indicator.

Highly sensitive and selective sensor for sunset yellow based on molecularly imprinted polydopamine-coated multi-walled carbon nanotubes.

Polydopamine (PDA) can be formed by monomeric self-polymerization in water. This convenient behavior was exploited to prepare a molecularly imprinted polymer (MIP) layer on the surface of multi-walled carbon nanotubes (MWCNTs) with sunset yellow (SY) as a template molecule. The prepared nanocomposites were characterized, and their electrochemical behavior towards SY was investigated. Under the optimized conditions, a glassy carbon electrode modified with the imprinted nanocomposite showed a highly selective and ultrasensitive electrochemical response to SY compared with the performance of control electrodes and previously reported electrochemical sensors for SY. The improved behavior of the developed sensor can be attributed to its superficial highly matched imprinted cavities on the excellent electrocatalytic matrix of MWCNTs and the electronic barrier of the non-imprinted PDA to outside molecules. The fabricated sensor expressed a linear relationship to SY concentrations from 2.2nM to 4.64µM with a detection limit of 1.4nM (S/N = 3). The sensor also exhibited excellent selectivity for SY over its structural analogs, good stability, and adequate reproducibility. The prepared sensor was successfully used to detect SY in real spiked samples. This methodology has potential application value and may be readily adapted to design other PDA-based MIP sensors.

[Negative HBsAg and HBsAb in Women of Child-bearing Age in Mianyang].

OBJECTIVES: To investigate the distribution of women with negative HBsAg and HBsAb at child-bearing age in Mianyang. METHODS: A total of 62 551 women aged 15-49 yr. were selected randomly using a multistage sampling strategy in Mianyang to participate in a questionnaire survey. Blood samples were collected during the survey. HBsAg and HBsAb were detected by enzyme-linked immunosorbent assay (ELISA). Those who were both HBsAg and HBsAb negative were deemed as susceptible to future infection of HBV. RESULTS: Data from 62 035 participants were valid for analysis: 28 460 (45.88%) were both HBsAg and HBsAb negative. Those aged 15-20 yr. had the lowest negative rate. Higher HBsAg and HBsAb negative rates were found in Han (lowest in Tibetan), rural residents, widowed/divorced (lowest in married), peasants (lowest in medical workers), those with a family history of Hepatitis B and without vaccination ( P<0.005). CONCLUSIONS: A high proportion of women at child-bearing age in Mianyang was found to be HBsAg and HBsAb negative. They should be monitored and vaccinated as a priority population in the prevention of mother-to-children infection of hepatitis B.

Trends, correlates, and disease patterns of antidepressant use among elderly persons in Taiwan.

PURPOSE: The population-based National Health Insurance database was used to investigate the trends, correlates, and disease patterns for elderly people in Taiwan who use antidepressants. METHODS: The National Health Research Institute provided a database of 1000,000 random subjects for study. We created a sample of subjects who were older than 65 years from 1997 to 2005. Trends, prevalence, and associated factors of antidepressant use were detected. We also examined the proportion of antidepressant use for psychiatric and medical disorders. RESULTS: The one-year prevalence of antidepressant use in elderly persons increased from 5.8 % in 1997 to 9.8 % in 2005. The one-year prevalence rates of tricyclic antidepressant (TCA), selective serotonin reuptake inhibitor (SSRI), serotonin-norepinephrine reuptake inhibitor (SNRI), serotonin modulator, and other antidepressant use in 2005 were 5.3, 2.6, 0.4, 2.9, and 0.6 %, respectively. Overall antidepressant use was higher for those in the 75- to 84-year-old age group, females, and those with higher Charlson Comorbidity Index scores. Among subjects using TCAs, 77.6 % users did not have a psychiatric diagnosis. Psychiatric disorders were commonly found in most SSRI and SNRI users (85.1 and 90.1 %, respectively). Subjects using SSRIs and SNRIs had higher proportions of psychiatric disorders such as neurotic depression, major depression, senile and presenile organic psychotic conditions, and anxiety. CONCLUSION: The prevalence of antidepressant use among elderly persons increased greatly from 1997 to 2005. SSRIs, SNRIs, and other antidepressants were used mostly by subjects with psychiatric disorders, whereas TCAs were used mostly by subjects with nonpsychiatric disorders.

Lin28B is an oncofetal circulating cancer stem cell-like marker associated with recurrence of hepatocellular carcinoma.

By using an expressed sequence tag bioinformatic algorithm, we identified that Lin28 homolog B (Lin28B) may have an oncofetal expression pattern which may facilitate detecting cancer cells in adults. It is also reported to be a potential marker for cancer stem cells. Therefore, we sought to verify oncofetal-stemness characters of Lin28B and test its potential as a circulating cancer stem cell-like marker in adult HCC patients. Lin28B mRNA was examined in a panel of fetal tissue, adult tissue and tumors. Lin28B was over-expressed or knocked down in HepG2 cells to evaluate its potential as a stem cell-like marker. RT-qPCR for Lin28B was performed in the peripheral blood mononuclear cells from patients with HCC receiving surgery (n=96) and non-HCC controls (n=60) and analyzed its clinical significance. Lin28B showed an oncofetal expression pattern. Its overexpression could upregulate stemness markers (OCT4, Nanog and SOX2) and enhance tumorsphere formation in vitro. Lin28B knockdown had opposite effects. Circulating Lin28B was detected in peripheral blood mononuclear cells in 3 cases (5%) of non-HCC controls and 32 cases (33.3%) of HCC patients. In HCC patients, circulating Lin28B was associated with high tumor grade (P=0.046), large size (P=0.005), high AJCC stage (P=0.044) and BCLC stage (P=0.017). Circulating Lin28B was significantly associated with decreased recurrence-free survival (P<0.001). Circulating Lin28B separated early stage HCC into 2 recurrence-free survival curves (P=0.003). In multivariate analysis, circulating Lin28B was an independent variable associated with early recurrence (P=0.045) and recurrence in early stage HCC (P=0.006). In conclusion, the oncofetal gene Lin28B is a potential oncofetal cancer-stem-cell-like circulating tumor cell marker that correlates with HCC recurrence after hepatectomy. Circulating Lin28B could refine early AJCC stages. Our finding supports the possible use of a TNMC (C for circulating tumor cells) staging system in HCC.

[Effects of bromocriptine on the treatment of prolactinomas in male patients and its influences on their sexual function].

OBJECTIVE: To explore the effect of bromocriptine in the treatment of male patients with prolactinoma and its impacts on their sexual function. METHODS: The clinical data of 29 male patients with prolactinomas treated with Bromocriptine were analysed, International Index of Erectile Function-5 (IIEF-5) was used to assess the sexual function of married patients before and after the treatment of Bromocriptine. RESULTS: The main clinical symptoms of male patients with prolactinomas were sexual dysfunction, headache and hypopsia, which were released significantly at 6 months after Bromocriptine therapy, with the decrease of serum prolactin (PRL) level (P < 0.05) and the improvement of basal testosterone (T) level (P < 0.05). The total normalization rate of PRL was 82.8%, and total effective rate of Bromocriptine therapy was 100%. According to the assessment of IIEF-5, all the male patients had their sexual function improved in various degree. CONCLUSION: Bromocriptine can improve the clinical symptoms of male patients with prolactinoma and their sexual function.

[Isolation and preliminary identification of stem-like cells in pituitary adenoma].

OBJECTIVE: To isolate and identify the pituitary adenoma stem-like cells from pituitary adenoma tissue. METHODS: Total RNA was prepared with 42 cases of pituitary adenoma tissue samples frozen in liquid nitrogen, the expression of Nestin was detected by RT-PCR. Tumor spheres were cultured in serum-free conditions and the immunefluorescence were used to detect the expression of stem cell markers such as Nestin, Sox2. RESULTS: The positive rate of Nestin mRNA expression was 88.1% (37/42). The cultured tumor spheres were found positive for Nestin, Sox2 by immunefluorescence detection. CONCLUSION: Stem cell markers are expressed in the pituitary adenoma tissue and the pituitary adenoma stem-like cells can be cultured in serum-free condition.

Trends, correlates, and disease patterns of antipsychotic use among children and adolescents in Taiwan.

PURPOSE: We used Taiwan's population-based National Health Insurance database to investigate the trends, correlates, and disease patterns of antipsychotic use among children and adolescents. METHODS: The National Health Research Institutes provided a database of 1,000,000 random subjects for study. We chose subjects who were aged 18 years or younger during 1997-2005. In this sample, subjects who were given at least one antipsychotic prescription, including first-generation antipsychotics (FGAs) or second-generation antipsychotics (SGAs), were identified. Trends, prevalence, and associated factors of antipsychotic use were determined. The proportion of antipsychotic use for psychiatric and medical disorders was also analyzed. RESULTS: The 1-year prevalence of SGA use increased from 0.00 % in 1997 to 0.09 % in 2005, whereas the 1-year prevalence of FGA use ranged from 2.24 to 3.43 % during this same period, with no significant change. Age and male gender were associated with higher SGA use. Among SGA users, the greatest proportion suffered from psychiatric disorders, including tics, hyperkinetic syndrome of childhood, schizophrenia, affective disorders, and autism. Among FGA users, a larger proportion was for medical conditions, including diseases of the digestive and respiratory systems. CONCLUSION: The prevalence of pediatric SGA use increased greatly from 1997 to 2005. Among pediatric subjects using antipsychotics, SGAs were mostly used for psychiatric disorders, whereas FGAs were mostly prescribed for medical conditions. Future research will focus on indication, dosage, frequency, duration, adverse effects, and off-label use of antipsychotics in the pediatric population.

Trends, correlates, and disease patterns of antidepressant use among children and adolescents in Taiwan.

The authors used a population-based database to investigate antidepressant use among children and adolescents in Taiwan. The National Health Research Institutes provided a database of 1 000 000 random subjects for study. The authors adopted this sample of subjects who were younger than 18 years during 1997 to 2005. Subjects with at least 1 antidepressant prescription were identified. Trends, prevalence, associated factors, and disease patterns of antidepressant use were detected. The 1-year prevalence of pediatric antidepressant use increased from 0.27% in 1997 to 0.47% in 2005. The 1-year prevalence of tricyclic antidepressant, selective serotonin reuptake inhibitor, and other antidepressant use among pediatric population was 0.23%, 0.20%, and 0.08%, respectively, in 2005. The prevalence of pediatric antidepressant use increased from 1997 to 2005. Among pediatric subjects with antidepressant use, selective serotonin reuptake inhibitors, and other antidepressants were used the most for psychiatric disorders, whereas tricyclic antidepressant was used the most for nonpsychiatric disorders.

[Repair and reconstruction of diaphragmatic defect during transcranial pituitary adenoma surgery].

OBJECTIVE: To investigate the application value of titanium-clip and artificial menigeal in the sella diaphragm repair and reconstruction during transcranial pituitary adenoma surgery. METHODS: Thirty eight patients undergoing transcranial pituitary adenoma surgery received sella diaphragmatic defect repair and reconstruction after the removal of tumor, intraoperative diaphragmatic defect was classified as three grades, titanium-clip, artificial menigeal and EC glue were used for the sella diaphragmatic repair and reconstruction. The short-term and long-term complications were compared with those in the control group which contains 74 patients without diaphragmatic reconstruction. RESULTS: As 14 to 46 months' follow-up in the diaphragmatic reconstruction group, there were no leakage of cerebrospinal fluid(CSF), infections, empty sella syndrome (ESS), optic nerve damage, and any other severe complications was found. Only one inter-sella hemorrhage without optic nerve damage was found 3 hours after surgery by CT scan. In the group without diaphragmatic reconstruction, more secondary empty sella, cerebrospinal fluid rhinorrhea and intra-cranial infection were found after the following transsphenoidal surgery (P < 0.05), there was one case underwent the 2nd time transcranial surgery 1 hours later due to severe inter-sella hemorrhage resulting in vision damage and conscious disturbance. There was no significant difference in pituitary function and vision improvement between the two groups as the occurrence of diabetes insipidus (P > 0.05). CONCLUSION: It is important to repair the sellae diaphragm defect during transcranial pituitary adenoma surgery, which could reduce short-term and long-term complication.