Predictive model for deep venous thrombosis caused by closed lower limb fracture after thromboprophylactic treatment.

OBJECTIVE: Currently, there are still no convincing clinical models predicting closed lower extremity fracture-associated deep vein thrombosis in patients treated through thromboprophylactic methods. We aimed at using two retrospective cohorts to develop and externally verify a clinical prediction model for deep vein thrombosis in patients treated with anticoagulants after suffering closed lower extremity fractures. PATIENTS AND METHODS: We evaluated the patients' pre- and post-operatively, to accurately determine the predictive power of the biomarkers and clinical risk factors. Two retrospective cohorts were used for the development and external verification of a pre-operative clinical prediction model (development: n = 2,253; verification: n = 833) and post-operative clinical prediction model (development: n = 1,422; verification: n = 449), respectively. RESULTS: The C-indices were used to show the predicted incidence of objective thrombosis at the pre- and post-operative stage, which were then compared with the observed incidence of thrombosis in both cohorts. Biomarkers and clinical indicators were included in pre- and post-operative nomograms, which were adequately calibrated in both cohorts. The cross-validated C-indices of the pre- and post-operative clinical prediction models in the verification cohort were 0.706 (95% Cl, 0.67-0.74) and 0.875 (95% Cl, 0.84-0.91), respectively. CONCLUSIONS: We present our findings of novel pre- and post-operative nomograms for the prediction of deep venous thrombosis in patients who received thromboprophylaxis after suffering closed lower extremity fractures.

Wernekinck Commissure Syndrome with Holmes Tremor: A Report of Two Cases and Review of Literature.

Wernekinck commissure syndrome is a rare midbrain infarction, it consists of several symptoms including bilateral cerebellar ataxia, ophthalmoplegia, and palatal tremor. Holmes tremor is a rare clinical syndrome characterized by a combination of resting, postural, and action tremors. We describe two cases of Wernekinck commissure syndrome with Holmes tremor. To the best of our knowledge, it has been rarely reported in the literature to date. Both of the cases were presented with acute onset of bilateral cerebellar ataxia, dysarthria, and Holmes tremor. In the treatment, one patient was given "clonazepam and benheisol," the other was received acupuncture therapy, both of them showed a marked improvement in ataxia and tremor.

High accuracy measurement of the ground-state hyperfine splitting in a 113Cd+ microwave clock: erratum.

We present an erratum to our Letter [Opt. Lett.40, 4249 (2015)OPLEDP0146-959210.1364/OL.40.004249]. This erratum corrects the nuclear Lande factor gI in Eq. (2). After correcting the error, the final ground-state hyperfine splitting frequency of the 113Cd+ ion is determined to be 15199862855.0287(10) Hz.

Spatiotemporal distribution, source apportionment and combined pollution of antibiotics in natural waters adjacent to mariculture areas in the Laizhou Bay, Bohai Sea.

The spatiotemporal distribution, source apportionment and combined pollution of 14 antibiotics in natural waters adjacent to mariculture farms of Laizhou Bay in the Bohai Sea were studied. The contribution proportion and quantity of each potential pollution source to antibiotics in natural water bodies were quantitatively described. The correlations between heavy metals and antibiotics and their underlying mechanisms in natural and aquaculture water environment were analyzed. Fourteen antibiotics were detected in natural water and sediment in the coastal area of Laizhou Bay. The maximum concentrations of sulfamethazine and trimethoprim in water reached tens or even hundreds of µg/L in winter. Trimethoprim was the main antibiotic in natural water bodies in winter and summer, and enrofloxacin was the principal antibiotic in sediments. Enrofloxacin, ciprofloxacin and oxytetracycline were detected in all underground water samples; thus, control of these antibiotics needs to be made a priority to mitigate groundwater contamination. PCA-MLR revealed that the potential sources of antibiotics in natural waters of Laizhou Bay include the mariculture wastewater (18.3%), the domestic sewage (63.3%) and the livestock wastewater (18.4%). Therefore, the antibiotic burden of Laizhou Bay was principally from the domestic sewage. In natural water, the concentration of Cu was positively correlated with antibiotics, which might be related to the common sources, the competitive adsorption in sediments and the easy complexation characteristic of Cu and antibiotics. Positive correlations among antibiotics and heavy metals were observed in mariculture sediments, while negative relationships were observed in natural sediments.

Antibiotics in marine aquaculture farms surrounding Laizhou Bay, Bohai Sea: Distribution characteristics considering various culture modes and organism species.

This study mainly investigated the distribution characteristics and risk assessment of 14 antibiotics in typical marine aquaculture farms surrounding the Bohai Sea. The effects of various culture modes (outdoor pond culture, recirculating water culture, greenhouse pond culture, raft culture, cage culture and bottom sowing culture), and diverse cultured organism species such as fish (grouper, bass, pike and turbot), mollusk (oyster, scallop, conch and mussel) and sea cucumber on the distribution of antibiotics in different mariculture pond matrices (seawater, sediment/biofilm and organism) were studied. In addition, antibiotic pollution levels in various matrices (water, sediment, organism and feed) from different mariculture areas surrounding the Bohai Sea and the Yellow Sea were compared. The biofilm on the inner wall of greenhouse pond was more capable of accumulating antibiotics than the biofilm attached to the rope for raft culture and net for cage culture, and other culture sediments. The antibiotic concentration level in the culture matrices (water, sediment/biofilm and organism) was the highest under greenhouse pond culture mode, and that under the industrial recirculating water culture mode was the lowest. Antibiotic concentration in culture matrices of fish ponds was higher than that of sea cucumber ponds and mollusk ponds. The levels of antibiotics in water and sediment from marine aquaculture farms in Laizhou (Bohai Sea coast) were higher than those in Haiyang and Jimo (Yellow Sea coast). Enrofloxacin in turbot might cause considerable harm to human health, and the risk of antibiotics in other seafood could be ignored. Antibiotic ecological risks and resistance risks were generally low in water. Fluoroquinolones posed medium to high ecological risks in the natural receiving water around the mariculture farm. Trimethoprim and enrofloxacin showed relatively high antibiotic resistance risks in mariculture water and natural water, which might exert selective pressure on the bacterial community in the environment.

Clinical epidemiology and outcome of HIV-associated talaromycosis in Guangdong, China, during 2011-2017.

OBJECTIVES: Talaromycosis is an invasive mycosis endemic to Southeast Asia. This study aimed to investigate the epidemiology, clinical features and prognostic factors of HIV-associated talaromycosis in Guangdong, China. METHODS: We retrospectively evaluated HIV patients hospitalized with histopathology- or culture-confirmed talaromycosis between 2011 and 2017. Factors associated with poor prognosis were identified using logistic regression. RESULTS: Overall, 1079 patients with HIV-associated talaromycosis were evaluated. Both the number and prevalence of talaromycosis among HIV admissions increased from 125 and 15.7% in 2011 to 253 and 18.8% in 2017, respectively, reflecting the increase in HIV admissions. Annual admissions peaked during the rainy season between March and August. Common clinical manifestations included fever (85.6%), peripheral lymphadenopathy (72.3%), respiratory symptoms (60.8%), weight loss (49.8%), skin lesions (44.5%) and gastrointestinal symptoms (44.3%). Common laboratory abnormalities were hypoalbuminaemia (98.6%), anaemia (95.6%), elevated aspartate aminotransferase level (AST) (76.9%), elevated alkaline phosphatase level (55.8%) and thrombocytopenia (53.7%). The median CD4 count was 9 cells/µL. Talaromyces marneffei was isolated from blood and bone marrow cultures of 66.6% and 74.5% of patients, respectively. The rate increased to 86.6% when both cultures were performed concurrently. At discharge, 14% of patients showed worsening conditions or died. Leucocytosis, thrombocytopenia, elevated AST, total bilirubin, creatinine and azole monotherapy independently predicted poor prognosis. CONCLUSIONS: The incidence of HIV-associated talaromycosis has increased in Guangdong with the high HIV burden in China. Skin lesions were seen in less than half of patients. Induction therapy with azole alone is associated with higher mortality. Findings from this study should help to improve treatment of the disease.

[Long-term oncological outcomes of laparoscopic versus abdominal surgery in stage â a1 （LVSI+）-â b1 cervical cancer patients with different tumor size: a big database in China].

Objective: To compare the long-term oncological outcomes between laparoscopic and abdominal surgery in stage Ⅰa1 （lymph-vascular space invasion-positive, LVSI+）- Ⅰb1 cervical cancer patients with different tumor sizes. Methods: Based on the Big Database of Clinical Diagnosis and Treatment of Cervical Cancer in China (1538 project database), patients with stage Ⅰa1 （LVSI+）-Ⅰb1 cervical cancer who treated by laparoscopic or abdominal surgery were included. The 5-year overall survival (OS) and 5-year disease-free survival (DFS) between the two surgical approaches were compared under 1∶1 propensity score matching (PSM) in different tumor diameter stratification. Results: (1) A total of 4 891 patients with stage Ⅰa1 （LVSI+）-Ⅰb1 cervical cancer who underwent laparoscopy or laparotomy from January 1, 2009 to December 31, 2016 were included in the 1538 project database. Among them, 1 926 cases in the laparoscopic group and 2 965 cases in the abdominal group. There were no difference in 5-year OS and 5-year DFS between the two groups before matching. Cox multivariate analysis suggested that laparoscopic surgery was associated with lower 5-year DFS (HR=1.367, 95%CI: 1.105-1.690, P=0.004). After 1∶1 PSM matching, 1 864 patients were included in each group, and there was no difference in 5-year OS between the two groups （94.1% vs 95.4%, P=0.151）. While, the inferior 5-year DFS was observed in the laparoscopic group (89.0% vs 92.3%, P=0.004). And the laparoscopic surgery was associated with lower 5-year DFS (HR=1.420, 95%CI: 1.109-1.818, P=0.006). (2) In stratification analysis of different tumor sizes, and there were no difference in 5-year OS and 5-year DFS between the laparoscopic group and abdominal group in tumor size ≤1 cm, >1-2 cm and >2-3 cm stratification (all P>0.05). Cox multivariate analysis showed that laparoscopic surgery were not related to 5-year OS and 5-year DFS (P>0.05). In the stratification of tumor size >3-4 cm, there was no difference in 5-year OS between the two groups (P>0.05). The 5-year DFS in the laparoscopic group was worse than that in the abdominal group (75.7% vs 85.8%, P=0.025). Cox multivariate analysis suggested that laparoscopic surgery was associated with lower 5-year DFS (HR=1.705, 95%CI: 1.088-2.674, P=0.020). Conclusions: For patients with stage Ⅰa1 （LVSI+）-Ⅰb1 cervical cancer, laparoscopic surgery is associated with lower 5-year DFS, and the adverse effect of laparoscopic surgery on oncology prognosis is mainly reflected in patients with tumor size >3-4 cm. For patients with tumor sizes ≤1 cm, >1-2 cm and >2-3 cm, there are no difference in oncological prognosis between the two surgical approaches.

Distribution, combined pollution and risk assessment of antibiotics in typical marine aquaculture farms surrounding the Yellow Sea, North China.

This study focused on the distribution, combined pollution, potential source and risk assessment of 17 antibiotics in an aquaculture ecosystem surrounding the Yellow sea, North China. Antibiotics were detected in various matrices (seawater, sediment/biofilm, organism and feed) in different aquaculture modes (greenhouse and outdoor aquaculture) during the wet and dry seasons in coastal areas of Shandong province. The innovation points of the study were as follows: (1) To the best of our knowledge, this study was one of the few to investigate the occurrence and distribution of antibiotics in mariculture environments along the Yellow Sea coast; (2) Biofilms, a focus of the study, might act as a sink for antibiotics in the aquaculture ecosystem; and (3) The correlation of heavy metals and antibiotic concentrations was proved, which could correspondingly be used as an indicator for antibiotic concentrations in the studied area. The levels of antibiotics in water were observed to be relatively low, at the ng/L level. Trimethoprim was the most prevalent antibiotic, and was detected in all water samples. Oxytetracycline was detected at high concentrations in biofilms (up to 1478.29 ng/g). Moreover, biofilms exhibited a higher antibiotic accumulation capacity compared to sediments. Concentrations of oxytetracycline and doxycycline were high in feed, while other antibiotics were almost undetected. Tetracycline was widely detected and the concentration of enrofloxacin was highest in organisms. Correlation analysis demonstrated that environmental parameters and other coexisting contaminants (e.g. heavy metals) significantly affected antibiotic concentrations. In addition, the concentration of Zn was significantly correlated with the total antibiotic concentration and was proportional to several antibiotics in water and sediment (biofilm) samples (p < 0.01). High Mn concentrations were closely related to total and individual (e.g. sulfadiazine, sulfamethazine and enrofloxacin) antibiotic levels, which may result in the combined contamination of the environment. Antibiotics in estuaries and groundwater generally originated from aquaculture wastewater and untreated/treated domestic sewage. Most of the detected antibiotics posed no risk to the environment. Ciprofloxacin and enrofloxacin found in water may present high ecological and resistance risks, while the two antibiotics observed to accumulate in fish may pose a considerable risk to human health through diet consumption. All antibiotics detected in seafood were lower than the respective maximum residue limits. This study can act as a reference for the government for the determination of antibiotic discharge standards in aquaculture wastewater and the establishment of a standardized antibiotic monitoring and management system.

Mechanistic understanding of compression-compression fatigue behavior of functionally graded Ti-6Al-4V mesh structure fabricated by electron beam melting.

In recent years, mesh structures have attracted significant interest for structural and functional applications. However, the mechanical strength and energy absorption ability of uniform mesh structured materials degrade with density. To address this challenge, we propose the concept of functionally graded mesh structures. The objective of the proposed research is to fundamentally understand the compressive behavior of graded mesh structures. The compression-compression fatigue behavior of functionally graded Ti-6Al-4V mesh structure under identical bulk stress condition is studied here. During cyclic deformation, it was observed that the local stress distribution in the struts was not uniform because of inhomogeneous mechanical properties of the constituents. Fatigue cracks first initiated in the lowest strength constituent, and then propagated until structural failure occurred. However, no obvious damage was observed in other constituents during the entire process. In contrast with iso-strain state, the fatigue life of graded structure is mainly determined by the constituent with the lowest strength.

Preconditioning of adipose-derived stem cells by phosphodiesterase-5 inhibition enhances therapeutic efficacy against diabetes-induced erectile dysfunction.

BACKGROUND: Adipose-derived stem cells have been considered as a promising therapy for erectile dysfunction. However, the therapeutic efficacy of adipose-derived stem cell-based therapy requires improvement. OBJECTIVE: To determine whether the inhibition of phosphodiesterase type 5 in adipose-derived stem cells would improve stem cell therapy for rats with diabetes-induced erectile dysfunction. MATERIALS AND METHODS: A phosphodiesterase type 5 siRNA was incorporated into lentiviral vectors and transduced into adipose-derived stem cells. The mRNA and protein levels of phosphodiesterase type 5 were evaluated. Three days after transduction, the adipose-derived stem cell supernatant was collected to determinate the levels of insulin-like growth factor 1 and vascular endothelial growth factor. Streptozotocin-induced diabetic rat models were established and used for comparative analysis of 1- and 2-week treatment regimens with intracavernosal injection of adipose-derived stem cells or Lv-siPDE5-modified adipose-derived stem cells. RESULTS: Lv-siPDE5-ADSCs secreted more insulin-like growth factor 1 and vascular endothelial growth factor in supernatants than unmodified adipose-derived stem cells. Preconditioned Adipose-derived stem cells-treated diabetic rats showed consistently superior erectile function when compared with non-preconditioned adipose-derived stem cells after 2 weeks of treatment. Lv-siPDE5-ADSCs provided additional benefits in recovery of cavernous structures with rapid effects (1 week) when compared to plain adipose-derived stem cells. These features were associated with the significantly increased levels of insulin-like growth factor 1 and vascular endothelial growth factor in Lv-siPDE5-ADSC-treated diabetic rats. CONCLUSIONS: Adipose-derived stem cell therapy could serve as an alternate approach for diabetes-induced erectile dysfunction, albeit with a long onset period. In vitro preconditioning of adipose-derived stem cells could accelerate the functional and structural recovery in vivo, indicating that preconditioning by inhibition of phosphodiesterase type 5 may improve adipose-derived stem cells therapy following diabetes-induced erectile dysfunction.

Ground-to-satellite time and frequency synchronization link with active carrier phase compensation.

In this paper, a synchronization link between one ground station and one geostationary satellite is established. The ground station receives retransmitted signals from the satellite, measures phase delay along the propagation route, and actively compensates back to its sending signals, realizing real-time phase fluctuation compensation. The transmitted signal contains two frequencies to eliminate common-mode phase noise. The difference between their carrier phase delays is measured. Different modes of carrier phase variation are separated and compensated, achieving a remaining time jitter of ±200 ps. Major sources of error are analyzed, and potential methods for improvement are discussed. The proposed ground-to-satellite link and active compensation method has potential applications in frequency standard dissemination to remote receivers (including ground stations or satellites). These potential applications justify further study of this system.

Structural features associated with multiferroic behavior in the RX3(BO3)4 system.

The magnetoelectric effect in the RX3(BO3)4 system (R = Ho, Eu, Sm, Nd, Gd; X = Fe, Al) varies significantly with the cation R despite very similar structural arrangements. Our structural studies reveal a symmetry reducing tilting of the BO3 planes and of the FeO6 polyhedra in the systems exhibiting low magnetic field induced electric polarization. Neutron scattering measurements reveal a lack of magnetic ordering indicating the primary importance of the atomic structure in the multiferroic behavior of this system.

Neural correlates of unstructured motor behaviors.

OBJECTIVE: We studied the relationship between uninstructed, unstructured movements and neural activity in three epilepsy patients with intracranial electroencephalographic (iEEG) recordings. APPROACH: We used a custom system to continuously record high definition video precisely time-aligned to clinical iEEG data. From these video recordings, movement periods were annotated via semi-automatic tracking based on dense optical flow. MAIN RESULTS: We found that neural signal features (8-32 Hz and 76-100 Hz power) previously identified from task-based experiments are also modulated before and during a variety of movement behaviors. These movement behaviors are coarsely labeled by time period and movement side (e.g. 'Idle' and 'Move', 'Right' and 'Left'); movements within a label can include a wide variety of uninstructed behaviors. A rigorous nested cross-validation framework was used to classify both movement onset and lateralization with statistical significance for all subjects. SIGNIFICANCE: We demonstrate an evaluation framework to study neural activity related to natural movements not evoked by a task, annotated over hours of video. This work further establishes the feasibility to study neural correlates of unstructured behavior through continuous recording in the epilepsy monitoring unit. The insights gained from such studies may advance our understanding of how the brain naturally controls movement, which may inform the development of more robust and generalizable brain-computer interfaces.

Direct observation of magnetic contrast obtained by photoemission electron microscopy with deep ultra-violet laser excitation.

Magnetic circular dichroism (MCD) and magnetic linear dichroism (MLD) have been investigated in a photoemission electron microscopy (PEEM) system excited by a deep ultra-violet (DUV) laser (with λ = 177.3 nm and hυ = 7.0 eV) for the first time. High resolution PEEM magnetic images (down to 43.2 nm) were directly obtained on a (001)-oriented magnetic FePt film surface with a circularly-polarized light under normal incidence. Furthermore, a stepped Cr seeding layer was applied to induce the formation of large-area epitaxial FePt films with (001) and (111) two orientations, where MLD with large asymmetry was observed in the transition area of two phases. It demonstrates that DUV laser can be a powerful source for high resolution magnetic imaging in the laboratory in absence of synchrotron facilities.

[Clinical investigation of ultrasound-guided percutaneous nephrolithotomy accessed by SVOF-principle and two-step puncture techniques].

Objective: To evaluate the safety and effectiveness of ultrasound-guided percutaneous nephrolithotomy (PCNL) accessed by SVOF-principle and two-step puncture techniques. Methods: A total of 838 cases with upper urinary stones underwent percutaneous nephrolithotomy successfully accessed by ultrasound-guided between June 2007 and December 2015 at Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. Of all cases were divided in two groups: hydronephrosis calyces puncture group include 425 cases and SVOF-principle puncture group include 413 cases. The access establishment time, operation time, stone free rate (SFR), postoperative complications, and postoperative hospitalization time between the two groups we compared by t test or χ2 test. Results: Statistically significant differences were observed between hydronephrosis calyces puncture group and SVOF-principle puncturegroup in the first access establishment time ((16.5±8.4) minutes vs. (11.2±5.9) minutes, t=3.931, P=0.013), one-stage SFR (74.3% vs. 85.7%, χ2=16.868, P=0.000), postoperative hospitalization time ((6.4±2.1) days vs. (4.8±1.8)days, t=4.574, P=0.000), transfusion rate (7.1% vs. 2.9%, χ2=8.027, P=0.006), and embolization rate (3.3% vs. 1.0%, χ2=5.390, P=0.020). There were no statistically significant differences in operation time, total SFR, postoperative fever and sever infection between these two groups (all P>0.05). In both two groups, no serious complications such as peripheral organ injury and death occurred. Conclusions: PCNL accessed guided by ultrasound with SVOF-principle and two-step puncture techniques has advantages of quick puncture location, high stone free rate, fewer complications and fast recovery. This technique is an effective and safe treatment option for upper urinary stones and deserved promotion and application in clinic.

Note: An atomic self-sustaining magnetic gradiometer with a 1/τ uncertainty property based on Larmor precession.

We demonstrate an atomic magnetic gradiometer based on self-sustaining Larmor precession. By coherent optical pumping, we measure the phase of the Larmor precession directly and observe that the gradiometer shows a 1/τ improvement in magnetic field gradient uncertainty over time τ. Since the measurement gives frequency signals, the gradiometer can be easily implemented by mixing and filtering the different frequency signals from two adjacent magnetometers. A gradient sensitivity of 186 fT/Hz/cm-1) is realized, which is close to the shot-noise limit. In a noisy environment, the gradiometer can still maintain its 1/τ behavior by suppressing 90% of the common-mode noise. This method should be widely applicable to the measurement of magnetic field gradients owing to its simplicity and outstanding performance.

[Clinical application of combined hepatic artery resection and reconstruction in surgical treatment for hilar cholangiocarcinoma].

Objective: To clarify whether the surgical treatment for hilar cholangiocarcinoma combined with artery reconstruction is optimistic to the patients with hilar cholangiocarcinoma with hepatic artery invasion. Methods: There were 384 patients who received treatment in the First Affiliated Hospital to Army Medical University from January 2008 to January 2016 analyzed retrospectively. There were 27 patients underwent palliative operation, 245 patients underwent radical operation, radical resection account for 63.8%. Patients were divided into four groups according to different operation method: routine radical resection group(n=174), portal vein reconstruction group (n=47), hepatic artery reconstruction group (n=24), palliative group(n=27). General information of patients who underwent radical operation treatment was analyzed by chi-square test and analysis of variance. The period of operation time, blood loss, the length of hospital stay and hospitalization expenses of the radical operation patients were analyzed by one-way ANOVA. Comparison among groups was analyzed by LSD-t test. Results: The follow-up ended up in June first, 2016. Each of patients followed for 6 to 60 months, the median follow-up period was 24 months. 1-, 3-, and 5-year survival rates were 81.3%, 44.9% and 13.5% of routine radical operation group, and were 83.0%, 44.7% and 15.1% of portal vein reconstruction group, and were 70.8%, 27.7% and 6.9% of hepatic artery reconstruction group, respectively. And 1-, 3-, and 5-year survival rates of hepatic artery reconstruction group was lower than routine radical group and portal vein reconstruction group significantly (P<0.05). However, the rate of postoperative complications of the hepatic artery reconstruction group and the routine radical operation group and the portal vein reconstruction group were 62.5%(15/24), 55.3%(96/174) and 51.5%(24/47), respectively. There was no significant difference among them (P>0.05). The data shows that the ratio of lymphatic metastasis in hepatic artery reconstruction group (70.8%) is much higher than them in routine radical operation group (20.1%) and portal vein reconstruction group (19.1%) significantly (P<0.05). The presented data also indicate that hepatic artery resection prolongs survival time comparing with patients undergoing palliative therapy for hilar cholangiocarcinoma. Cox regression analysis indicate that hepatic artery resection and reconstruction is a protective factor compare with palliative therapy (RR=0.38, 95%CI: 0.22-0.67). The significant reason for shorter survival time is a positive correlation between hepatic artery invasion and lymph node metastasis. Conclusion: Hepatic artery resection and reconstruction has beneficial impact on oncologic long-term outcome in patients with advanced stage hilar cholangiocarcinoma.

Regulatory functions of trehalose-6-phosphate synthase in the chitin biosynthesis pathway in Tribolium castaneum (Coleoptera: Tenebrionidae) revealed by RNA interference.

RNA interference (RNAi) is a very effective technique for studying gene function and may be an efficient method for controlling pests. Trehalose-6-phosphate synthase (TPS), which plays a key role in the synthesis of trehalose and insect development, was cloned in Tribolium castaneum (Herbst) (TcTPS) and the putative functions were studied using RNAi via the injection of double-stranded RNA (dsRNA) corresponding to conserved TPS and trehalose-6-phosphate phosphatase domains. Expression analyses show that TcTPS is expressed higher in the fat body, while quantitative real-time polymerase chain reaction results show that the expression of four trehalase isoforms was significantly suppressed by dsTPS injection. Additionally, the expression of six chitin synthesis-related genes, such as hexokinase 2 and glutamine-fructose-6-phosphate aminotransferase, was suppressed at 48 and 72 h post-dsTPS-1 and dsTPS-2 RNA injection, which were two dsTPS fragments that had been designed for two different locations in TcTPS open reading frame, and that trehalose content and trehalase 1 activity decreased significantly at 72 h post-dsRNA injection. Furthermore, T. castaneum injected with dsTPS-1 and dsTPS-2 RNA displayed significantly lower levels of chitin and could not complete the molting process from larvae to pupae, revealing abnormal molting phenotypes. These results demonstrate that silencing TPS gene leads to molting deformities and high mortality rates via regulation of gene expression in the chitin biosynthetic pathway, and may be a promising approach for pest control in the future.

Connexin36 localization to pinealocytes in the pineal gland of mouse and rat.

Several cell types in the pineal gland are known to establish intercellular gap junctions, but the connexin constituents of those junctions have not been fully characterized. Specifically, the expression of connexin36 (Cx36) protein and mRNA has been examined in the pineal, but the identity of cells that produce Cx36 and that form Cx36-containing gap junctions has not been determined. We used immunofluorescence and freeze fracture replica immunogold labelling (FRIL) of Cx36 to investigate the cellular and subcellular localization of Cx36 in the pineal gland of adult mouse and rat. Immunofluorescence labelling of Cx36 was visualized exclusively as puncta or short immunopositive strands that were distributed throughout the pineal, and which were absent in pineal sections from Cx36 null mice. By double immunofluorescence labelling, Cx36 was localized to tryptophan hydroxylase-positive and 5-hydroxytryptamine-positive pinealocyte cell bodies and their large initial processes, including at intersections of those processes and at sites displaying a confluence of processes. Labelling for the cell junction marker zonula occludens-1 (ZO-1) either overlapped or was closely associated with labelling for Cx36. Pinealocytes thus form Cx36-containing gap junctions that also incorporate the scaffolding protein ZO-1. FRIL revealed labelling of Cx36 at ultrastructurally defined gap junctions between pinealocytes, most of which was at gap junctions having reticular, ribbon or string configurations. The results suggest that the endocrine functions of pinealocytes and their secretion of melatonin is supported by their intercellular communication via Cx36-containing gap junctions, which may now be tested by the use of Cx36 null mice.

Effects of mutant TDP-43 on the Nrf2/ARE pathway and protein expression of MafK and JDP2 in NSC-34 cells.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that affects motor neurons and lacks an effective treatment. The disease pathogenesis has not been clarified at present. Pathological transactive response DNA-binding protein 43 (TDP-43) plays an important role in the pathogenesis of ALS. Nuclear translocation of nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2) is found in a mutant TDP-43 transgenic cell model, but its downstream antioxidant enzyme expression is decreased. To elucidate the specific mechanism of Nrf2/ARE (antioxidant responsive element) signaling dysfunction, we constructed an ALS cell model with human mutant TDP-43 using the NSC-34 cell line to evaluate the impact of the TDP-43 mutation on the Nrf2/ARE pathway. We found the nuclear translocation of Nrf2, but the expression of total Nrf2, cytoplasmic Nrf2, and downstream phase II detoxifying enzyme (NQO1) was decreased in NSC-34 cells transfected with the TDP-43-M337V plasmid. Besides, TDP-43-M337V plasmid-transfected NSC-34 cells were rounded with reduced neurites, shortened axons, increased levels of intracellular lipid peroxidation products, and decreased viability, which suggests that the TDP-43-M337V plasmid weakened the antioxidant capacity of NSC-34 cells and increased their susceptibility to oxidative damage. We further showed that expression of the MafK protein and the Jun dimerization protein 2 (JDP2) was reduced in TDP-43-M337V plasmid-transfected NSC-34 cells, which might cause accumulation of Nrf2 in nuclei but a decrease in NQO1 expression. Taken together, our results confirmed that TDP-43-M337V impaired the Nrf2/ARE pathway by reducing the expression of MafK and JDP2 proteins, and provided information for further research on the molecular mechanisms of TDP-43-M337V in ALS.