Reorganization of intrinsic functional connectivity in early-stage Parkinson's disease patients with probable REM sleep behavior disorder.

REM sleep behavior disorder (RBD) symptoms in Parkinson's disease (PD) suggest both a clinically and pathologically malignant subtype. However, whether RBD symptoms are associated with alterations in the organization of whole-brain intrinsic functional networks in PD, especially at early disease stages, remains unclear. Here we use resting-state functional MRI, coupled with graph-theoretical approaches and network-based statistics analyses, and validated with large-scale network analyses, to characterize functional brain networks and their relationship with clinical measures in early PD patients with probable RBD (PD+pRBD), early PD patients without probable RBD (PD-pRBD) and healthy controls. Thirty-six PD+pRBD, 57 PD-pRBD and 71 healthy controls were included in the final analyses. The PD+pRBD group demonstrated decreased global efficiency (t = -2.036, P = 0.0432) compared to PD-pRBD, and decreased network efficiency, as well as comprehensively disrupted nodal efficiency and whole-brain networks (all eight networks, but especially in the sensorimotor, default mode and visual networks) compared to healthy controls. The PD-pRBD group showed decreased nodal degree in right ventral frontal cortex and more affected edges in the frontoparietal and ventral attention networks compared to healthy controls. Furthermore, the assortativity coefficient was negatively correlated with Montreal cognitive assessment scores in the PD+pRBD group (r = -0.365, P = 0.026, d = 0.154). The observation of altered whole-brain functional networks and its correlation with cognitive function in PD+pRBD suggest reorganization of the intrinsic functional connectivity to maintain the brain function in the early stage of the disease. Future longitudinal studies following these alterations along disease progression are warranted.

Ethylidene-Tethered Chromene-Pyrone Hybrids as Potential Plant-Growth Regulators from an Endolichenic Phaeosphaeria Species.

Phaeosphaeria sp., a lichen-associated fungus, produced six skeletally new dimeric spiciferones (1-6) and four known metabolites (7-10). The new structures were elucidated by spectroscopic analysis, and their absolute configurations were determined by electronic circular dichroism calculations. Compounds 1 and 3-6 represent the first examples of ethylidene-bridged dimers from the building blocks 4H-chromene-4,7(8H)-dione and α-pyrone, and 2 is a unique homodimer of spiciferone. Compounds 1, 2, and 5-9 significantly inhibited the growth of weed-like dicot Arabidopsis thaliana at 100.0 µM. Notably, 8 showed the strongest inhibitory activity against the fresh weight and root elongation of A. thaliana with the IC50 values of 32.04 and 26.78 µM, respectively, whereas 1, 8, and 9 stimulated the growth of A. thaliana at lower concentrations. Meanwhile, compounds 2 and 6 exhibited weak inhibitory effects on the root elongation of monocot rice, while 1 and 8 exhibited growth-promoting effects on the shoot and root elongation of rice in a roughly dose-dependent manner.

Chloroplast proteostasis: A story of birth, life, and death.

Protein homeostasis (proteostasis) is a dynamic balance of protein synthesis and degradation. Because of the endosymbiotic origin of chloroplasts and the massive transfer of their genetic information to the nucleus of the host cell, many protein complexes in the chloroplasts are constituted from subunits encoded by both genomes. Hence, the proper function of chloroplasts relies on the coordinated expression of chloroplast- and nucleus-encoded genes. The biogenesis and maintenance of chloroplast proteostasis are dependent on synthesis of chloroplast-encoded proteins, import of nucleus-encoded chloroplast proteins from the cytosol, and clearance of damaged or otherwise undesired "old" proteins. This review focuses on the regulation of chloroplast proteostasis, its interaction with proteostasis of the cytosol, and its retrograde control over nuclear gene expression. We also discuss significant issues and perspectives for future studies and potential applications for improving the photosynthetic performance and stress tolerance of crops.

Forensic Validation and Application Evaluation of IDentifier DNA Typing Kit (Yan-Huang34).

OBJECTIVES: To investigate the technical performance of IDentifier DNA typing kit (YanHuang34) and evaluate its forensic application value. METHODS: Following the Criterion of Forensic Science Human Fluorescence STR Multiplex Amplification Reagent (GB/T 37226-2018), IDentifier DNA typing kit (YanHuang34) was verified in 11 aspects of species specificity, veracity, sensibility, adaptability, inhibitor tolerance, consistency, balance, reaction condition verification, mixed samples, stability and inter batch consistency. The system efficiency of IDentifier DNA typing kit (YanHuang34) was compared with the PowerPlex® Fusion 6C System, VersaPlex® 27PY System and VeriFilerTM Plus PCR Amplification Kit. The IDentifier DNA typing kit (YanHuang34) was used to detect the swabs of biological samples in daily cases and the STR performances were observed. RESULTS: IDentifier DNA typing kit (YanHuang34) had good species specificity, veracity, adaptability, inhibitor tolerance and balance. The sensibility was up to 0.062 5 ng. It was able to detect different types of samples, degraded samples and inhibitor mixed samples. Complete DNA typing could be obtained for samples with the mixture ratio less than 4∶1. The system efficiency of IDentifier DNA typing kit (YanHuang34) was very high, with TDP up to 1-1.08×10-37, CPEtrio and CPEduo up to 1-5.47×10-14 and 1-6.43×10-9, respectively. For the touched biological samples in actual cases, the effective detection rate was 21.05%. The system efficiency of kinship, single parent and full sibling identifications was effectively improved. CONCLUSIONS: The IDentifier DNA typing kit (YanHuang34) is adaptive to the GB/T 37226-2018 requirements. It can be used for individual identification and paternity identification, and is suitable for application in the field of forensic science.

The effects of hyperglycemia on the biological behavior of human gingival fibroblasts on a titanium surface.

BACKGROUND: It is well known that diabetes mellitus is one of the high-risk factors for periodontitis and also for the failure of implant restorations. Usually, the success of an implant restoration depends on both the good osseointegration and the stable soft tissue interface on the implant neck. A good gingival interface of the implant neck is the barrier that enables implant to resist oral microorganisms and the site of initiation of peri-implantitis. This study sought to investigate the effects of hyperglycemia on the attachment and proliferation of human gingival fibroblasts (HGFs) on pure titanium surfaces. METHODS: HGFs were cultured in cell culture mediums with different glucose concentrations (i.e., 5.5, 8. 8, 10, and 15 mmol/L) for 7 d and seeded on pure titanium surfaces. The cells that were seeded on the titanium surfaces had been cultured in cell culture mediums with different glucose concentrations for 3 and 7 d. The attached HGFs on the titanium surfaces were counted for all groups using a blood cell counting plate, and the results were statistically analyzed. The morphologies of the attached HGFs on the titanium surfaces were observed for all the groups using a scanning electronic microscope. RESULTS: As the glucose concentrations increased, the number of attached HGFs on the titanium surfaces decreased. The numbers of attached cells in Groups A and B 7 d after being seeded on the titanium surfaces were more than those 3 d after being seeded (P<0.05). The numbers of attached cells in Groups C and D 3 d after being seeded on the titanium were more than those 7 d after being seeded (P<0.05). The scanning electronic microscope showed that the attached cells in Groups A and B proliferated well, and most cells grew one on top of another. Conversely, the attached cells in Groups C and D proliferated sparsely and the cell morphologies were not good. CONCLUSIONS: The attachment and proliferation of HGFs on pure titanium surfaces were inhibited by increases in glucose concentrations, and the inhibition was further enhanced by the passage of time.

Structurally Diverse Sesquiterpenoids with Anti-neuroinflammatory Activity from the Endolichenic Fungus Cryptomarasmius aucubae.

Two new sterpurane sesquiterpenoids named sterpurol D (1) and sterpurol E (2), and one skeletally new sesquiterpene, cryptomaraone (3), bearing a 5,6-fused bicyclic ring system, along with five known ones, sterpurol A (4), sterpurol B (5), paneolilludinic Acid (6), murolane-2α, 9ß-diol-3-ene (7) and (-)-10,11-dihydroxyfarnesol (8) were isolated from an endolichenic fungus Cryptomarasmius aucubae. The structures of the new compounds were elucidated by analysis of NMR spectroscopic spectra and HRESIMS data. The absolute configurations of 1 and 2 were established by spectroscopic data analysis and comparison of specific optical rotation, as well as the biosynthetic consideration. Additionally, compounds 1, 2, 4-6, and 8 showed significant nitric oxide (NO) production inhibition in Lipopolysaccharide (LPS)-induced BV-2 microglial cells with the IC50 values ranging from 9.06 to 14.81 µM.

Repetitive transcranial magnetic stimulation improves Parkinson's freezing of gait via normalizing brain connectivity.

Robust, effective treatments for Parkinson's freezing of gait remain elusive. Our previous study revealed beneficial effects of high-frequency rTMS over the supplementary motor area. The present study aims to explore the neural mechanisms of rTMS treatments utilizing novel exploratory multivariate approaches. We first conducted a resting-state functional MRI study with a group of 40 Parkinson's disease patients with freezing of gait, 31 without freezing of gait, and 30 normal controls. A subset of 30 patients with freezing of gait (verum group: N = 20; sham group: N = 10) who participated the aforementioned rTMS study underwent another scan after the treatments. Using the baseline scans, the imaging biomarkers for freezing of gait and Parkinson's disease were developed by contrasting the connectivity profiles of patients with freezing of gait to those without freezing of gait and normal controls, respectively. These two biomarkers were then interrogated to assess the rTMS effects on connectivity patterns. Results showed that the freezing of gait biomarker was negatively correlated with Freezing of Gait Questionnaire score (r = -0.6723, p < 0.0001); while the Parkinson's disease biomarker was negatively correlated with MDS-UPDRS motor score (r = -0.7281, p < 0.0001). After the rTMS treatment, both the freezing of gait biomarker (0.326 ± 0.125 vs. 0.486 ± 0.193, p = 0.0071) and Parkinson's disease biomarker (0.313 ± 0.126 vs. 0.379 ± 0.155, p = 0.0378) were significantly improved in the verum group; whereas no significant biomarker changes were found in the sham group. Our findings indicate that high-frequency rTMS over the supplementary motor area confers the beneficial effect jointly through normalizing abnormal brain functional connectivity patterns specifically associated with freezing of gait, in addition to normalizing overall disrupted connectivity patterns seen in Parkinson's disease.

High-frequency rTMS over the supplementary motor area improves freezing of gait in Parkinson's disease: a randomized controlled trial.

INTRODUCTION: Freezing of gait (FOG) contributes to falls in Parkinson's disease (PD), but robust, effective treatments remain elusive. There is evidence indicating that the supplementary motor area (SMA) plays an important role in the pathogenesis of FOG and may therefore be a potential neuromodulation target. The present study explored the clinical efficacy of high-frequency repetitive transcranial magnetic stimulation (rTMS) over the SMA on FOG in PD patients. METHODS: A group of 30 PD patients with FOG were enrolled in a randomized, double-blind, sham-controlled trial. Patients were randomly allocated 2:1 to receive ten sessions of either real (N = 20) or sham (N = 10) 10 Hz rTMS over SMA. The patients were assessed at baseline (T0), after the 5th (T1) and 10th (T2) sessions, and then 2 weeks (T3) and 4 weeks (T4) after the last session. The primary clinical outcome was the Freezing of Gait Questionnaire score (FOGQ), with the Movement Disorder Society-Unified Parkinson's Disease Rating Scale motor scores (MDS-UPDRS III) and Timed Up and Go test as secondary clinical outcomes. All the assessments were carried out at the "ON" state. RESULTS: With a four week's follow-up, there were significant interaction effects in the FOGQ (effect of group*time, p = 0.04), MDS-UPDRS III (p = 0.02) and several gait variables (total duration, p < 0.01; cadence, p = 0.04; turn duration, p = 0.01; and turn to sit duration, p = 0.02). Post-hoc analyses revealed a significantly decreased FOGQ score at T2 and T4, and significant improvements of MDS-UPDRS III and gait variables at T1, T2, T3 and T4 in the rTMS group. No significant improvements were found in the sham group. CONCLUSION: High-frequency rTMS over SMA may ultimately serve as an add-on therapy for alleviating FOG in PD patients.

Wide Awake Secondary Tendon Reconstruction.

The wide awake anesthesia technique is a useful tool in secondary tendon reconstruction. With active participation of the patient, the tendon repair can be adjusted appropriately to prevent repairs that are too tight or too loose. Areas of tendon scarring or triggering can be identified and released. Other advantages of active participation include reduction of gapping, ensuring adequate strength of repair, and avoiding tendon imbalances. Last, it allows intraoperative patient education and may therefore increase patient satisfaction. This article discusses how the technique can be applied to tenolysis, 2-stage tendon reconstruction, and tendon transfer.

MicroRNA­100­5p inhibits osteoclastogenesis and bone resorption by regulating fibroblast growth factor 21.

MicroRNAs (miRNAs/miRs) are post­transcriptional regulators that serve important roles in osteoclastogenesis and bone metabolism; however, the roles of miRNAs have not been completely clarified. The present study aimed to investigate the effects of miR­100­5p on the mechanism of liver­bone endocrine metabolism. A miRNA microarray analysis was conducted to evaluate the miRNA expression profile during receptor activator of nuclear factor­κB ligand­stimulated osteoclastogenesis. Hematoxylin and eosin and tartrate­resistant acid phosphatase staining were performed to analyze the trabecular bone microstructure and osteoclast differentiation. The mRNA and protein expression levels were assessed by reverse transcription­quantitative polymerase chain reaction and western blotting, respectively. The results revealed that in vitro osteoclast differentiation and in vivo bone resorption were suppressed by miR­100­5p overexpression. In vivo, a decrease in miR­100­5p and an increase in FGF21 were simultaneously observed in mice following ovariectomy (OVX). Bioinformatics analysis and experimental data confirmed that FGF21 was a direct target of miR­100­5p. Conversely, augmentation of miR­100­5p using a specific agomir in OVX­operated mice decreased the levels of FGF21 in the serum and liver, and prevented osteoclastogenesis and bone loss. The present study revealed that FGF21 may be a signal molecule associated with the mechanism of liver­bone endocrine metabolism and may be targeted by miR­100­5p. In addition, miR­100­5p may serve an important role in protecting against OVX­induced osteoporosis.

Author Correction: Altered resting-state brain activity in Parkinson's disease patients with freezing of gait.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.

Altered resting-state brain activity in Parkinson's disease patients with freezing of gait.

Freezing of gait (FOG) is a common and debilitating symptom in Parkinson's disease (PD). The current study investigated alterations of resting-state spontaneous brain activity in PD patients with FOG. A total of 29 patients with FOG, 28 patients without FOG and 31 controls were included. All subjects underwent resting-state functional MRI, and the amplitude of low-frequency fluctuation (ALFF) was calculated to measure the spontaneous brain activity. Between-group differences and correlations with FOG severity (both subjective and objective measures) were analyzed. Compared to those without FOG, patients with FOG showed increased ALFF in right anterior cingulate cortex (ACC) and left inferior parietal lobule (IPL), as well as decreased ALFF in right superior frontal gyrus (SFG), bilateral cerebellum and left thalamus. Correlation analyses demonstrated that ALFF within the right SFG, right ACC and bilateral pallidum were positively correlated with FOG; while ALFF within the thalamus, putamen, cerebellum and sensorimotor regions were negatively correlated. Our results indicate that FOG is associated with dysfunction within frontal-parietal regions, along with increased inhibitory outputs from basal ganglia. Additionally, altered activity of cerebellum implicates its role in the pathophysiology of FOG. These findings provide further insight into the underlying neural mechanisms of FOG in PD patients.

Levodopa Effect on Basal Ganglia Motor Circuit in Parkinson's Disease.

AIMS: To investigate the effects of levodopa on the basal ganglia motor circuit (BGMC) in Parkinson's disease (PD). METHODS: Thirty PD patients with asymmetrical bradykinesia and 30 control subjects were scanned using resting-state functional MRI. Functional connectivity of the BGMC was measured and compared before and after levodopa administration in patients with PD. The correlation between improvements in bradykinesia and changes in BGMC connectivity was examined. RESULTS: In the PD-off state (before medication), the posterior putamen and internal globus pallidus (GPi) had decreased connectivity while the subthalamic nucleus (STN) had enhanced connectivity within the BGMC relative to control subjects. Levodopa administration increased the connectivity of posterior putamen- and GPi-related networks but decreased the connectivity of STN-related networks. Improvements in bradykinesia were correlated with enhanced connectivity of the posterior putamen-cortical motor pathway and with decreased connectivity of the STN-thalamo-cortical motor pathway. CONCLUSION: In PD patients with asymmetrical bradykinesia, levodopa can partially normalize the connectivity of the BGMC with a larger effect on the more severely affected side. Moreover, the beneficial effect of levodopa on bradykinesia is associated with normalization of the striato-thalamo-cortical motor and STN-cortical motor pathways. Our findings inform the neural mechanism of levodopa treatment in PD.

Cost-effectiveness of microsurgical reconstruction for head and neck defects after oncologic resection.

BACKGROUND: Microvascular free tissue transfer has become the main technique used for head and neck reconstruction. We assessed the cost-effectiveness of free flap reconstruction for head and neck defects after oncologic resection for squamous cell carcinoma (SCC). METHODS: We developed a Markov model of the cost, quality of life, survival, and incremental cost-effectiveness of reconstruction with free tissue transfer compared with locoregional flaps. Health state probabilities and quality of life scores were determined from literature. Costs were determined from institutional experience. Outcomes included quality-adjusted life years, costs, and incremental cost-effectiveness ratio. RESULTS: Free flap reconstruction was more costly than pedicled flap but associated with greater quality of life with no survival benefit. A value <$50,000 per quality-adjusted life-year (QALY) was defined as cost-effective. The incremental cost-effectiveness for head and neck free flap reconstruction was below the threshold and, therefore, free flap reconstruction is cost-effective. Reconstruction was more cost-effective for patients with lower stage cancers: $4643 per QALY for stage I SCC, $8226 for stage II, $17,269 for stage III, and $23,324 for stage IV. Univariate sensitivity analysis showed the cost-effectiveness would remain <$50,000 for all stages of SCC for all variables except for QALY after locoregional reconstruction without complications. CONCLUSION: Microsurgical head and neck reconstruction is cost-effective compared with locoregional flaps, even more so in patients with early-stage cancer. This finding supports the current practice of free flap head and neck reconstruction. Screening and early detection are important to optimize costs. © 2016 Wiley Periodicals, Inc. Head Neck 39: 541-547, 2017.

The study of brain functional connectivity in Parkinson's disease.

Parkinson's disease (PD) is a neurodegenerative disorder primarily affecting the aging population. The neurophysiological mechanisms underlying parkinsonian symptoms remain unclear. PD affects extensive neural networks and a more thorough understanding of network disruption will help bridge the gap between known pathological changes and observed clinical presentations in PD. Development of neuroimaging techniques, especially functional magnetic resonance imaging, allows for detection of the functional connectivity of neural networks in patients with PD. This review aims to provide an overview of current research involving functional network disruption in PD relating to motor and non-motor symptoms. Investigations into functional network connectivity will further our understanding of the mechanisms underlying the effectiveness of clinical interventions, such as levodopa and deep brain stimulation treatment. In addition, identification of PD-specific neural network patterns has the potential to aid in the development of a definitive diagnosis of PD.

A rabbit pulmonary vein myocyte isolation method based on simultaneous heart and pulmonary vein perfusion.

Myocytes in the pulmonary veins (PV) play a pivotal role in the development of paroxysmal atrial fibrillation (AF). It is therefore important to understand physiological characteristics of these cells. Studies on these cells are, however, markedly impeded by the fact that single PV myocytes are very difficult to obtain due to lack of effective isolation methods. In this study, we described a novel PV myocyte isolation method. The key aspect of this method is to establish a combination of retrograde heart perfusion (via the aorta) and anterograde PV perfusion (via the pulmonary artery). With this simultaneous perfusion method, a better perfusion of the PV myocytes can be obtained. As results, the output and viability of single myocytes isolated by simultaneous heart and PV perfusion method were increased compared with those in conventional retrograde heart perfusion method.

Whole mitochondrial genome sequence and mutations of the hypertension model inbred rat strain (Muridae; Rattus).

We reported the complete mitochondrial genome sequencing of a important hypertension model inbred rat strain for the first time. The total length of the mitogenome was 16,310 bp. It harbored 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and 1 non-coding control region. The mutation events contained in this strain were also reported.

[Expression of CSN Complex in ATRA-induced APL Cell Differentiation and Its Clinical Significance].

OBJECTIVE: To investigate the expression of CSN complex (COP9 signal some subunits) in the patients with acute promyelocytic leukemia (APL) and its significance in the ATRA-induced APL differentiation. METHODS: Using the NB4 cells as a model, morphologic observation and myeloid differentiation marker CD11b detection were used to monitor ATRA-induced APL differentiation, the expression of CSN complex in cell differentiation was detected by Western blot and reverse transcription real time fluorescent quantitative PCR (RT-qPCR) method. RT-qPCR was also used to detect the relative expression level of COP9 signalosome subunits in the APL patients and remission after treatment. RESULTS: ATRA could obviously enhance CD11b expression; the cell morphology showed obvious differentiation characteristics. During the differentiation, the expression of COP9 signalosome subunits was down-regulated by ATRA. Meanwhile, the CSN expression level in newly diagnosed APL patients was much higher than that in controls (non-leukemia) (P < 0.05). The level of CSN expression was obviously down-regulated when APL patients achieved complete remission. CONCLUSION: The high CSN expression level in APL patients can be down-regulated by ATRA. CSN complex may have a significant effect on the pathogenesis and therapy of APL.

[Analysis and Discussion on Calculating Likelihood Ratio of DNA Mixture].

OBJECTIVE: To analyze and discuss four methods of calculating likelihood ratio of DNA mixture. METHODS: In the case with CNAS-T0757 proficiency testing in 2013, the likelihood ratios were calculated and compared among four methods, including unrestricted combinatorial method, Clayton's method, p2 principle method, and recommendations from ISFG. RESULTS: The likelihood ratios were maximum by Clayton's method and recommendations from ISFO, followed by result of the unrestricted combinational method. The minimum likelihood ratio was obtained by p2 principle. CONCLUSION: The unrestricted combinational method could give fUrthest consideration to both information preservation and appraiser protection.

[Dipylidium caninum infection in an infant: one case report].

This paper reports the diagnosis and therapy of one case of Dipylidium caninum infection in an infant.