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Alzheimer's disease (AD) is a common neurodegenerative disorder characterized by the accumulation of misfolded amyloid-β (Aβ) peptides, which is caused by an imbalance between Aβ production and clearance. Picroside II (Picr II), the principal active constituent of Picrorhizakurroa Royle ex Benth, possesses a range of pharmacological properties, such as anti-inflammatory, antioxidant, and antiapoptotic effects. However, the exact role of Picr II in AD is not yet fully understood. In this study, we investigated the effects of Picr II on AD and found that it inhibited amyloid precursor protein (APP) processing by enhancing autophagy rather than through the ubiquitin-proteasome pathway, resulting in a reduction in Aβ production. Furthermore, Picr II treatment improved the decline in autophagy, which was due not only to an increase in autophagic flux but also to an increase in the fusion of autophagosomes with lysosomes. Most importantly, daily treatment with Picr II (20 mg/kg, i.p.) throughout the experiment rescued the learning and memory in Aβ-treated mice. Taken together, these results suggest that Picr II may have a potent neuroprotective effect against Aβ-induced memory decline by enhancing autophagy, indicating that Picr II may be a promising therapeutic agent for AD.

Previous studies have provided solid evidence of social contagion under the item memory task as individuals would unconsciously incorporate the information that originated from their collaborators. Nonetheless, it is unknown whether social contagion also exists in source memory, another branch of episodic memory. Moreover, it remains controversial whether stimuli emotionality influences the degree to which individuals engage in social contagion. To address these issues, the current experiment adopted the modified social contagion paradigm and 90 participants were recruited to learn different emotional valence words of varied sharing levels (e.g., shared, partly shared, and unshared) and font source (e.g., Chinese Song and Kai typeface) at the encoding stage, then under the item and source retrieval tasks participants successively carried out a deep communication of a triad collaborative recall memory and followed individual memory. We monitored how collaborative memory affected subsequent individual memories including the items and their sources that were initially only encoded by collaborators. Three main findings emerged. (a) The social contagion not limited to item memory, even existed in source memory, strengthening the source monitoring framework and the dual-process model. (b) The emotionality of stimuli including positive and negative ones had a significant effect on individual contagious episodic memory, the phenomenon was creatively named the Emotionality Induced Social Contagion Effect (EISC). Notably, under the two retrieval tasks the social contagion of positive and negative stimuli was similar, but both were significantly higher than that of neutral ones, consistently showing a U-shape curve. (c) The social contagion was also sensitive to item types and font typeface, especially for partly shared and Song typeface items, along with a three-way interaction among item types, font typeface, and stimuli emotionality.

IntroductionInfantile spasms (IS) is a type of severe epileptic encephalopathy that occurs in infancy and early childhood. IS is characterised clinically by epileptic spasms, often accompanied by sleep disorder and abnormal circadian rhythm. The endogenous circadian rhythm disorder, in turn, can make spasms worse. Melatonin has also been found to have anticonvulsant and neuroprotective properties by adjusting the circadian rhythm. However, there are lack of relevant studies on controlling IS by using melatonin. This study aims to analyse the therapeutic effect of melatonin supplementation for the treatment of IS.Methods and analysisThis is a triple-blinded (trial participant, outcome assessor and the data analyst), prospective, randomised controlled trial to be conducted in the Department of Paediatrics, The First Medical Center of Chinese PLA General Hospital, Beijing, China from November 2020. Patients (n=70) aged 3 months to 2 years with IS will be recruited in this study after receiving written consent from their parents or guardians. Patients will be randomly divided into two equal groups and treated with a combination of adrenocorticotropic hormone, magnesium sulfate and either melatonin or placebo. Clinical data from the patients in the two groups before and after the treatment will be collected and compared. The primary outcome will be assessed 2 weeks later by seizure diaries and reported as the average reduced rate of spasms frequency. Secondary outcomes include the response rate (the rate of spasms-free), electroencephalogram hypsarrhythmia assessment and the psychomotor development assessment (Denver Developmental Screening Test). Sleep quality and safety will also be assessed.Ethics and disseminationThe protocol for this study was approved by the Ethics Committee of Chinese PLA General Hospital (reference number S2020-337-01) and was reported according to the Standard Protocol Items: Recommendations for Interventional Trials statement. Findings of this research will be disseminated through national and international meetings, conferences and peer-reviewed journals.Trial registration numberChiCTR2000036208.

High density, fine pitch wafer level bonding technology is necessary for heterogeneous integration. However, underfill would be a challenging when bonding pitch decreases. Hybrid bonding technology with metal-to-metal and passivation-to-passivation bonding simultaneously was considered as a feasible solution. In this work, a hybrid bonding scheme with asymmetric embedded Cu-Sn/Cu bumps structure was investigated. Photosensitive BCB used as passivation materials was coated on only one side of bonding pairs. This asymmetric structure allows for precise alignment. BCB curing and Cu-Sn Solid-liquid-diffusion bonding were both achieved under the temperature of 250°C. The thickness of BCB and metal was concisely designed considering the volume shrinking of polymer during bonding process. To achieve void free hybrid bonding interface, efforts were mainly paid to some key aspects. Surface pre-treatment were firstly introduced to ensure no oxidation and residual polymer on the surface of bumps before bonding. A combination of plasma and formic gas treatment were adopted. Furthermore, considering coefficients of thermal expansion (CTE) mismatch of BCB and embedded bumps, a compatible bonding process with well-matched temperature and stress was proposed and optimized to get a void-free bonding interface under the guidance of independent variable test result. Bonding interface microstructure was observed and mechanical test, electrical test of Kelvin and daisy-chain structure were performed to evaluate bonding quality, also reliability test which included HTST and TCT had been carried out. The results confirmed that embedded bump hybrid bonding would be a convincing technology for future 3D integration.

Metal-dielectric hybrid bonding is a critical technique of inter-chip connections for 3D integration. In this work, a chip-level embedded hybrid bonding structure using Cu-Sn/BCB Bonding is proposed and realized. This embedded microbump structure with photosensitive BCB coated on the single side could be successfully developed without chemical mechanical polishing (CMP), while the cure temperature (250°C) of BCB is the same as that of Cu-Sn thermal compression bonding or solid-liquid interdiffusion (SLID). The deviation of misalignment could be reduced effectively due to the embedded bonding structure. Consecutive and void-free interface could be got at BCB-metal and BCB-Si boundary with optimized scale design of BCB patterns while the diameter/pitch of microbumps is 50μm/100μm, the thickness of bonding layer is approximately 10μm. Also, key factors of process for realizing good quality of bonding is discussed. The electrical test shows that interface of metal microbumps could be improved further. Besides, the die shear test shows good mechanical strength of this hybrid bonding structure as the shear strength of hybrid bonding area and pure BCB bonding area are over 13MPa and 19MPa, respectively.

The Aristaless-related homeobox protein (ARX) is a transcription factor expressed in the developing forebrain, skeletal muscle, pancreas, testis, and a variety of other tissues. It is known to have context-dependent transcriptional activator and repressor activity, although how it can achieve these opposing functions remains poorly understood. We hypothesized phosphorylation status might play a role in pivoting ARX between functioning as an activator or repressor. To gain further mechanistic insight as to how ARX functions, we identified multiple phosphorylation sites on ARX. We further established PKA as the kinase that phosphorylates ARX at least at Ser266 in mice. Two other kinases, CK2α and CDK4/cyclin D1, were also identified as kinases that phosphorylate ARX in vitro. Unexpectedly, phosphorylation status did not change either the nuclear localization or transcriptional function of ARX.

In millimeter-wave devices, Fan-out Wafer Level Package for system integration is a popular trend to meet the requirements of high performance, small dimensions and low cost. Compared to conventional Fan-out Wafer Level Package with molding compound, embedded Silicon Fan-out (eSiFO) Wafer Level Package has become an attractive technology due to low cost, less warpage problem and fine-pitch redistribution layer (RDL) production capability. However, silicon substrate is usually of low resistivity, which limits the application of eSiFO technology in millimeter-wave field. Thus, high efficiency and low loss transmission line has become an important topic in eSiFO. In this paper, a design of coplanar waveguide (CPW) with shield line for eSiFO in millimeter-wave applications is proposed. Electrical modeling of transmission line structure was carried out by HFSS. The CPW with and without shield line were fabricated compatible with semiconductor processes. Measurement and test results shows the shield line can provide good impedance matching, suppress undesirable substrate loss, as well as decrease crosstalk from adjacent lines.

Objective Mutations in phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) complex have been associated with a broad spectrum of brain and organ overgrowth syndromes. For example, mutations in phosphatidylinositol-3-kinase regulatory subunit 2 (PIK3R2) have been identified in human patients with megalencephaly polymicrogyria polydactyly hydrocephalus (MPPH) syndrome, which includes brain overgrowth. To better understand the pathogenesis of PIK3R2-related mutations, we have developed and characterized a murine model. Methods We generated a knock-in mouse model for the most common human PIK3R2 mutation, p.G373R (p.G367R in mice) using CRISPR/Cas9. The mouse phenotypes, including brain size, seizure activity, cortical lamination, cell proliferation/size/density, interneuron migration, and PI3K pathway activation, were analyzed using standard methodologies. For human patients with PIK3R2 mutations, clinical data (occipitofrontal circumference [OFC] and epilepsy) were retrospectively obtained from our clinical records (published / unpublished). Results The PI3K-AKT pathway was hyperactivated in these mice, confirming the p.G367R mutation is an activating mutation in vivo. Similar to human patients with PIK3R2 mutations, these mice have enlarged brains. We found cell size to be increased but not cell numbers. The embryonic brain showed mild defects in cortical lamination, although not observed in the mature brain. Furthermore, electroencephalogram (EEG) recordings from mutant mice showed background slowing and rare seizures, again similar to our observations in human patients. Interpretation We have generated a PIK3R2 mouse model that exhibits megalencephaly and EEG changes, both of which overlap with human patients. Our data provide novel insight into the pathogenesis of the human disease caused by PIK3R2 p.G373R mutation. We anticipate this model will be valuable in testing therapeutic options for human patients with MPPH. ANN NEUROL 2020;88:1077-1094.

Aim: To explore the independent prognostic value of STK24 expression in terms of overall survival and recurrence-free survival and the potential mechanisms of its dysregulation in non-small-cell lung adenocarcinoma. Patients & methods: Data were from the Cancer Genome Atlas-lung adenocarcinoma. Results: Increased STK24 expression was an independent prognostic indicator of unfavorable overall survival (Hazard ratio: 1.478; 95% CI: 1.149–1.901; p

We made head-shaped Cu-Sn solder joints with diameters about 10 μm, 20 μm, and 50 μm and reflowed the samples at 240 °C, 260 °C, and 280 °C for 60 s to 600 s. In the 10 μm bumps, there is only one Cu6Sn5 grain after reflow, thus the classic model of scallop-type grain growth of Cu6Sn5 does not apply. Also, the grain growth in the small size bumps has the faster growth rate. We proposed a surface diffusion-controlled model to explain the new kinetics. According to our model, the diffusion activation energy is calculated to be QS = 0.18 ± 0.02 eV/atom and diffusion frequency factor to be DS0 = 5.65 × 10−3 cm2/s.

Early brain development requires a tight orchestration between neural tube patterning and growth. How pattern formation and brain growth are coordinated is incompletely understood. Previously we showed that aristaless-related homeobox (ARX), a paired-like transcription factor, regulates cortical progenitor pool expansion by repressing an inhibitor of cell cycle progression. Here we show that ARX participates in establishing dorsoventral identity in the mouse forebrain. In Arx mutant mice, ventral genes, including Olig2, are ectopically expressed dorsally. Furthermore, Gli1 is upregulated, suggesting an ectopic activation of SHH signaling. We show that the ectopic Olig2 expression can be repressed by blocking SHH signaling, implicating a role for SHH signaling in Olig2 induction. We further demonstrate that the ectopic Olig2 accounts for the reduced Pax6 and Tbr2 expression, both dorsal specific genes essential for cortical progenitor cell proliferation. These data suggest a link between the control of dorsoventral identity of progenitor cells and the control of their proliferation. In summary, our data demonstrate that ARX functions in a gene regulatory network integrating normal forebrain patterning and growth, providing important insight into how mutations in ARX can disrupt multiple aspects of brain development and thus generate a wide spectrum of neurodevelopmental phenotypes observed in human patients.

Importin 13 (IPO13) is a key member of the importin β superfamily, which can transport cargoes both into and out of the nucleus to contribute to a variety of important cellular processes. IPO13 is known to undergo phosphorylation, but the impact of this on function has not been investigated. Here, we show for the first time that IPO13 is phosphorylated by cAMP-dependent protein kinase A specifically at serine 193. Results from fluorescence recovery after photobleaching and fluorescence loss in photobleaching approaches establish that negative charge at serine 193 through phosphorylation or point mutation both reduces IPO13 nuclear import and increases its nuclear export. Importantly, phosphorylation also appears to enhance cargo interaction on the part of IPO13, with significant impact on localization, as shown for the Pax6 homeobox-containing transcription partner. This is the first report that IPO13 can be phosphorylated at Ser193 and that this modification regulates IPO13 subcellular localization and nucleocytoplasmic transport function, with important implications for IPO13's role in development and other processes.

Dynamin-related protein 1 (Drp1) is a key regulator of mitochondrial fission. Our previous studies proved that the inhibition of Drp1 may help attenuate traumatic brain injury (TBI)-induced functional outcome and cell death through maintaining normal mitochondrial morphology and inhibiting activation of apoptosis. However, the molecular mechanisms of Drp1 after TBI remain poorly understood. In this study, we investigated the role of mitochondrial division inhibitor 1 (Mdivi-1), a small molecule inhibitor of Drp1, in underlying mechanisms of general autophagy and mitochondria autophagy (mitophagy) after experimental TBI. In vivo, we found that autophagosomes accumulated in cortical neurons at 24h after TBI, owing to the enhanced autophagy indicated by the accumulation of LC3 and the decrease of p62; but Mdivi-1 reversed the enhancement. Mdivi-1 also alleviated the number of LC3 puncta and TUNEL-positive structures in cells, indicating that autophagy maybe involved in Mdivi-1's anti-apoptosis effects. Then, the expression level of mitochondrial dynamics related and mitophagy related proteins was assessed using the isolated mitochondria. The results showed that TBI-induced mitochondrial fission (represented by Drp1), mtDNA concentration down-regulation and PTEN induced putative kinase 1 (PINK1)-Parkin mediated mitophagy activation were all inhibited by Mdivi-1. In addition, TBI-induced blood-brain barrier (BBB) disruption and matrix metalloproteinases (MMP)-9 expression up-regulation were inhibited following Mdivi-1 treatment. In vitro, Mdivi-1 significantly alleviated the scratch injury-induced cell death, loss of mitochondrial membrane potential, reactive oxygen species (ROS) production and ATP reduction in primary cortical neurons (PCNs). Additionally, the lysosome inhibitor chloroquine (CQ) abrogated the Mdivi-1-induced decrease in autophagosomes accumulation and cell death at 24h both in the basal state and under the conditions of scratch cell injury. Together, these data demonstrate that Mdivi-1 mitigates TBI-induced BBB disruption and cell death at least in part by a mechanism involving inhibiting autophagy dysfunction and mitophagy activation.

Ca2+/calmodulin-dependent protein kinase IIα (CaMKIIα) is identified as a Ca2+-dependent kinase in brain involved in the activation of Tryptophan hydroxylase (TPH) acting through direct phosphorylation of TPH, and playing key roles in the signaling pathways initiated by various G protein-coupled 5-HT receptors. The goal of this study is to detect whether there are changes of CaM and CaMKIIα in dorsal raphe nucleus in the rats exposed to single-prolonged stress (SPS), which is a model employed in post-traumatic stress disorder (PTSD) study extensively. A total of 90 male Wistar rats were randomly divided into a normal control group and SPS groups of 7d, 14d. The changes of CaM/CaMKIIα were detected by immunohistochemistry, reverse transcription-polymerase chain reaction and western blot. Our results demonstrate that both expressions of CaM and CaMKIIα significantly increase (P < 0.001) in the SPS 7d group than that in the control group, and then decreased dramatically (P < 0.001) 14 days after SPS. Our results confirm that SPS induce changes of CaM/CaMKIIα in the dorsal raphe nucleus. Changes of CaM/CaMKIIα may be associated with the activation of 5-HT1A receptor, and may contribute to the progress of molecular mechanism of PTSD.

To evaluate the therapeutic effect and safety of rapamycin in treatment of children with tuberous sclerosis complex (TSC) complicated with epilepsy.This was an open-label, prospective, self-controlled study. From Sep. 2011 to Sep. 2013, 52 patients with the diagnosis of tuberous sclerosis complicated with epilepsy receiving rapamycin treatment for at least 24 weeks were enrolled.Of the 52 children, 34 were male and 18 female. The median age at onset of epilepsy was 4.8 months (4 days-49 months), the median age for treatment with rapamycin was 27 months (4.5-172.5 months). Ten children had a family history of TSC. In 24 children TSC gene detection was carried out, among whom TSC1 mutation was detected in 4 cases and TSC2 mutation in 20. Before rapamycin therapy, 59.62%, (31/52) patients took more than 3 antiepileptic drugs, of whom 10 cases even took more than 5 kinds of antiepileptic drugs. Fifty-two patients received rapamycin treatment for 24 weeks, seizure free rate was 25.00% (13 cases), the total effective rate was 73.08% (38 cases); 31 cases received treatment for 48 weeks, seizure free 6 cases, total effective 23 cases; 17 cases accepted treatment for 72 weeks, seizure free 5 cases, total effective 13 cases; 12 cases received treatment for 96 weeks, seizure free 3 cases, total effective 9 cases. With the decrease of seizure attacks, use of antiepileptic drug types were reduced simultaneously, they had a negative correlation. Before rapamycin therapy, the average frequency of seizures was 70.27 times/d, the number of antiepileptic drug kinds was 1.30. After 24, 48, 72, 96 weeks' treatment, the average seizure frequency was reduced to 1.94-2.80 times /d and the antiepileptic drugs were reduced to 0.83-0.97 kinds. On every visit during the follow-up, blood and urine routine tests, liver and kidney function test showed no abnormality in the 52 cases. The drug dosage was 1 mg/(m(2)×d), average 0.7 mg/d (0.35-1.20 mg/d). Blood concentrations of rapamycin remained below 10 µg/L (average 6.5 µg/L). The main side effect was oral ulcer which happened in 23.08% (12/52). The oral ulcer would disappeared 2-3 days later. 17.31% (9/52 cases) had upper respiratory infection.Rapamycin was effective in children with tuberous sclerosis and epilepsy with few adverse reactions. The daily dose of rapamycin for children patients is 1 mg/m(2), which has a certain effect on seizures and a good safety profile.

The authors sought to investigate the clinical features and characteristics of genetic mutation in patients with aspartylglucosaminuria.Clinical data of two pediatric siblings in a family were analyzed retrospectively and relative literature was reviewed in order to study the clinical features, imaging and enzymatic characteristics and genetic mutations.Case 1, the proband, male, he was hospitalized at 20 months of age because of fever and hepatosplenomegaly for nine days. This child was of moderate nutritional status and normal development. Blood tests showed hemoglobin 78.0 g/L, RBC3.18 × 10¹²/L, WBC 4.06 × 10⁹/L, neutrophils 0.236, lymphocytes 0.631, platelets 34 × 10⁹/L, C-reactive protein 17 mg/L. Blood biochemistry showed alanine aminotransferase 67.1 U/L, aspartate aminotransferase 74.1 U/L, serum albumin 32.8 g/L, direct bilirubin 10.5 µmol/L, lactate dehydrogenase 301.7 U/L. Bone marrow cytology showed reactive morphological changes in bone marrow cells. Atypical lymphocytes could be seen in both peripheral blood and bone marrow smears. Cranial MRI showed poor myelination. Aspartylglucosaminidase activity in peripheral leucocytes of the proband 5.7 nmol/(g × min) vs. normal control26.6 nmol/(g × min). On his AGA gene and that of his parents, a heterozygous mutation site located in exon 3, c.392CT (p.S131L), was identified as a novel mutation inherited from his father. The mutation from his mother has not been detected. The proband was not responsive to the anti-infectious medication, nutritional intervention and symptomatic treatment.He died one month after diagnosis.His elder brother, Case 2, showed fever, recurrent respiratory tract infection and progressive psychomotor regression with hepatosplenomegaly from the age of four years. Cranial MRI revealed extensive symmetrical leukodystrophy in bilateral cerebra, cerebellum and brainstem.He died at the age of six years.Related literature was summarized, and no Chinese AGU cases had been reported; 221 foreign cases were collected. The clinical and imaging characteristics were summarized. Delay in language development was one of the clinical symptoms that the majority of parents of AGU children first noted.Patients with aspartylglucosaminuria lack of specific symptoms.For children with unexplained delayed speech and progressive mental retardation, the possibility of AGU should be considered, and efforts be made for enzymatic and genetic diagnosis. c.392CT (p.S131L) was identified as a novel mutation of AGA gene.