Background:Guillain-Barré syndrome (GBS) is a severe acute autoimmune neuropathy with limited established therapeutic options. Efgartigimod, a human IgG antibody Fc fragment, can increase IgG degradation, which thus may be a promising therapeutic medicine for GBS. The aims of this study were to evaluate the clinical effectiveness and safety of efgartigimod in GBS patients. Methods:A retrospective study was conducted on GBS patients at the first affiliated hospital of Zhengzhou university and Nanyang central hospital from February 2024 to June 2025. The medical records of patients received IVIg, efgartigimod, or IVIg sequential efgartigimod (ISE) were reviewed. Disease severity was evaluated using GBS disability score (GBS-DS) and Inflammatory Neuropathy Cause and Treatment (INCAT) disability score at baseline, week1, week2, week3, week4, and the final follow-up. The primary outcome was the proportion of good improvement, defined as a reduction of at least 2 points in the GBS-DS score compared to the baseline. All adverse events occurred during the treatment period were documented. Results:52 patients were enrolled and received IVIg (n=20), efgartigimod (n=16), or ISE (n=16). Good improvement was higher in the efgartigimod group than IVIg group both at week 2 (37.5% vs. 0%, p<0.01) and the final visit (81.3% vs. 40.0%, p=0.02). Throughout follow-up, the proportion with GBS-DS ≤1 in the efgartigimod group was consistently higher compared to the other two groups. Although not statistically significant, the mean time to reach GBS-DS ≤1 was shorter in the efgartigimod group (2.6± 1.2 weeks) compared to the IVIg (3.1 ± 1.1 weeks), In the ISE group, the average time to achieve GBS-DS ≤ 1 was 2.8 ± 1.2 weeks. Regarding safety, the incidence of treatment-related adverse events (TEAEs) was lower in the efgartigimod group18.8% (3/16) than in the IVIg50.0% (10/20) and ISE 43.8% (7/16) groups. Conclusions:Based on the preliminary finding, the retrospective real-world data suggesting a potential signal of benefit for efgartigimod in GBS, with no new safety concerns identified. However, this study cannot establish comparative efficacy given the methodological limitations, and large-scale well-controlled head-to-head trials remain imperative.

The benefits of intravenous thrombolysis in patients with acute minor stroke remain controversial. For the aim of providing a better therapeutic strategy, high-quality trials are required to validate the efficacy of thrombolytic medicine other than intravenous recombinant tissue plasminogen and tenecteplase. In the trial, we evaluate the efficacy and safety of urokinase (UK) in acute minor stroke. This multicenter, open-label, blinded-endpoint, randomized controlled clinical trial enrolled patients with minor stroke within 6 h of symptom onset, with a NIHSS score ≤ 5. The trial was conducted at 25 hospitals in China between October 2020 and February 2023. Eligible patients were randomized to the UK group (1,000,000 U) or the best medicine treatment group. The responsible investigator recommended and implemented the best medicine treatment based on guidelines. The primary endpoint was an excellent functional outcome, defined as a modified Rankin scale (mRS) score of 0–1 at 90 days. The primary safety outcome was symptomatic intracranial hemorrhage (sICH) within 36 h. A total of 999 patients were enrolled in the trial, the median age was 64 years, 371 (36.9

Alzheimer's disease (AD) is the most common cause of dementia, with global case numbers projected to reach 153 million in 2050 1 . AD is highly heritable, with twin-based heritability estimates of 60-80% 2 . While 1,200 causal loci are predicted to exist for AD 3 , approximately 80 have been associated with AD in two recent studies 4,5 , suggesting that many loci remain to be discovered 6 . Here, we analyzed data from 183,620 AD cases and 2.6 million controls from diverse ancestries, identifying 118 loci in a multi-ancestry analysis and 9 additional loci in ancestry-specific analyses, 48 of which are new. We identified new AD risk genes, prioritized potential drug targets, and identified microglia and, for the first time, several neuronal cell types enriched for AD-associated genetic risk. Moreover, we improved polygenic prediction and estimated a single-nucleotide polymorphism (SNP) heritability of 19%. Together, our findings offer insights into the genetic architecture and potential pathobiology of AD, as well as specific targets for future drug development research.

Background: Intravenous thrombolysis is increasingly used in patients following acute minor ischaemic stroke, but its benefit is not yet clear. Urokinase as the first-generation intravenous thrombolytic drug, which was recommend UK for intravenous thrombolysis in ischaemic stroke patients in China guidelines. However, the efficacy and safety of UK for acute minor ischaemic stroke remains unknown.Methods: This multicentre, open-label, blinded-endpoint, randomised clinical trial was conducted across 25 hospitals in China. Patients who presented with minor ischaemic stroke (National Institutes of Health Stroke Scale [NIHSS] ≤5) within 6 h of symptom onset were randomly assigned (1:1) to receive intravenous thrombolysis (1,000,000 U UK dissolved in 100 mL of saline, given as a continuous intravenous infusion over 30 min) or antiplatelet therapy. The primary endpoint was an excellent functional outcome, defined as a score of 0–1 on the modified Rankin scale (mRS) at 90 days. The primary safety outcome was symptomatic intracranial haemorrhage (sICH) within 36 h. The endpoints were assessed in a blinded manner. The TRUST trial is registered with ClinicalTrials.gov, NCT 04420351.Findings: Between October 2020 and November 2022, 999 participants were enrolled (503 patients were assigned to the UK intravenous thrombolysis group, and 496 patients were assigned to the oral antiplatelet therapy group). At 90 days, the primary endpoint occurred in 427 patients (84.9%) in the UK group and 425 patients (85.7%) in the antiplatelet group (adjusted risk ratio [RR] 1.00, 95%CI 0.96–1.05). Three UK-treated patients (0.6%) vs. 1 antiplatelet-treated patient (0.2%) had sICH (RR 1.83, 95%CI (0.16–20.27).Interpretation: For patients with minor acute ischaemic stroke presenting within 6 h of symptom onset, treatment with UK was not found to be superior to antiplatelet therapy on excellent functional outcome at 90 days. The risk of sICH did not differ between the two treatment groups.Trial Registration: The TRUST trial is registered with ClinicalTrials.gov, NCT 04420351.Funding: The trial was funded by the National Key R&D Program of China (2017YFC1308202).Declaration of Interest: All authors declare no competing interests.Ethical Approval: Approved by the ethics committees of the First Affiliated Hospital of Zhengzhou University and of each participating site. Written informed consent for participation in the trial was provided by the patients or their representatives. The trial was analysed by independent academic statisticians following a protocol and statistical analysis plan available in the appendix.

Objective To investigate the efficacy and safety of ofatumumab in the treatment of neuromyelitis optica spectrum disorders (NMOSDs). Methods The clinical data of 25 patients with NMOSDs hospitalized in The First Affiliated Hospital of Zhengzhou University from April 2022 to March 2023 and treated with ofatumumab were retrospectively analyzed. They were divided into 2 groups, Group A with 12 cases receiving regular medication, and Group B with 13 cases receiving medication based on the percentage of CD19+B lymphocytes. Annual relapse rate (ARR), the number of relapses, the frequency and time of relapses, and the symptoms of relapses were calculated before and at one year of medication. The neurological function was assessed by Expanded Disability Status Scale (EDSS) before and at one year of medication. The adverse drug reactions were recorded during medication. Results After one year treatment with ofatumumab, 3 patients (12%) experienced recurrence. One patient in Group A experienced recurrence twice, one month and 5 months after treatment, respectively. At the second recurrence, the percentage of CD19+B lymphocytes was 2.80%. Two patients in Group B experienced recurrence, one patient relapsed twice at 2 and 6 months after medication. The percentage of CD19+B lymphocytes was 0.80% and 0.21%, respectively; another patient relapsed once at 2 months after medication. At the time of recurrence, the percentage of CD19+B lymphocytes was 0.70%. There was a statistically significant difference in ARR before and after one year treatment (F = 29.061, P = 0.000). After one year's treatment, the ARR of Group A (t = 13.215, P = 0.001) and Group B (t = 19.259, P = 0.000) was decreased than before. There were 8 cases of the injection site pain, 3 cases of fever after injection, and one case of headache after injection. There were no serious adverse reactions such as disability and death, and EDSS scores were not increased. Conclusions Continuous administration of ofatumumab every 3 months or based on the percentage of CD19+B lymphocyte can reduce recurrence. Combination with corticosteroids should be alert to the risk of infection.

The extracellular matrix plays a crucial role in the growth of human neural stem cells (hNSCs) by forming a stem cell niche, bothin vitroandin vivo. The demand for defined synthetic substrates has been increasing recently in stem cell research, reflecting the requirements for precise functions and safety concerns in potential clinical approaches. In this study, we tested the adhesion and expansion of one of the most representative hNSC lines, the ReNcell VM Human Neural Progenitor Cell Line, in a pure-synthesized short peptide-basedin vitroniche using a previously established integrin-binding peptide array. Spontaneous cell differentiation was then induced using two differentin vitroapproaches to further confirm the multipotent features of cells treated with the peptides. Twelve different integrin-binding peptides were capable of supporting hNSC adhesion and expansion at varied proliferation rates. In the ReNcell medium-based differentiation approach, cells detached in almost all peptide-based groups, except integrinα5β1 binding peptide. In an altered differentiation process induced by retinoic acid containing neural differentiation medium, cell adhesion was retained in all 12 peptide groups. These peptides also appeared to have varied effects on the differentiation potential of hNSCs towards neurons and astrocytes. Our findings provide abundant options for the development ofin vitroneural stem cell niches and will help develop promising tools for disease modeling and future stem cell therapies for neurological diseases.

The discovery of skin intranuclear inclusions and GGC repeat expansion of NOTCH2NLC has greatly promoted the diagnosis of neuronal intranuclear inclusion disease (NIID). With highly heterogeneous clinical manifestations, NIID patients tend to be underdiagnosed at early stages. This study comprehensively studied clinical manifestations, magnetic resonance imaging (MRI), and peripheral nerve conduction in 24 NIID and 166 other neurodegenerative disease (ND) subjects. The nomogram was plotted using the “rms” package, and the t-distributed stochastic neighbor embedding algorithm was performed. Associations between skin intranuclear inclusions and NOTCH2NLC GGC repeats were further analyzed. The clinical, MRI, and peripheral nerve conduction features seriously overlapped in NIID and ND patients; they were assigned variables according to their frequency and specificity in NIID patients. A nomogram that could distinguish NIID from ND was constructed according to the assigned variables and cutoff values of the above features. The occurrence of skin intranuclear inclusions and NOTCH2NLC GGC repeats ≥ 60 showed 100

Amyloid-β (Aβ) plays an important role in the neuropathology of Alzheimer's disease (AD), but some factors promoting Aβ generation and Aβ oligomer (Aβo) neurotoxicity remain unclear. We here find that the levels of ArhGAP11A, a Ras homology GTPase-activating protein, significantly increase in patients with AD and amyloid precursor protein (APP)/presenilin-1 (PS1) mice. Reducing the ArhGAP11A level in neurons not only inhibits Aβ generation by decreasing the expression of APP, PS1, and β-secretase (BACE1) through the RhoA/ROCK/Erk signaling pathway but also reduces Aβo neurotoxicity by decreasing the expressions of apoptosis-related p53 target genes. In APP/PS1 mice, specific reduction of the ArhGAP11A level in neurons significantly reduces Aβ production and plaque deposition and ameliorates neuronal damage, neuroinflammation, and cognitive deficits. Moreover, Aβos enhance ArhGAP11A expression in neurons by activating E2F1, which thus forms a deleterious cycle. Our results demonstrate that ArhGAP11A may be involved in AD pathogenesis and that decreasing ArhGAP11A expression may be a promising therapeutic strategy for AD treatment.

Both specific and innate immune responses play important roles in autoimmune encephalitis (AE). We aimed to explore the predictive value of the systemic inflammation index (SII) at admission as a peripheral biomarker of treatment response of AE. A total of 146 patients diagnosed with AE in the First Affiliated Hospital of Zhengzhou University from January 1, 2018 to September 22, 2020 were retrospectively and consecutively analyzed as per the inclusion criteria and divided into two groups according to their response to immunotherapy after 30 days. The predictive value of the SII as a peripheral biomarker for AE treatment response was calculated using the receiver operating characteristic curve analysis, which showed that the best SII cut-off value for predicting poor response to AE treatment was 863.3; the area under the curve was 0.75, with 83.0% sensitivity and 72.0% specificity. The risk factors for poor response to AE treatment were analyzed; univariable analysis showed that the rate of decreased level of consciousness, rate of cognitive or mental behavior abnormality, cerebrospinal fluid pressure, blood neutrophils, platelets, time until treatment initiation, neutrophil to lymphocyte ratio, platelet to lymphocyte ratio, and SII were significantly higher in patients with poor response to AE immunotherapy after 30 days than in patients with good response. Meanwhile, the blood lymphocyte counts and Glasgow Coma Scale (GCS) scores in patients with poor response were significantly lower than those in patients with good response (all p < 0.05), and multivariable binary logistic regression with backward stepwise method showed that decreased levels of consciousness, time until treatment initiation and SII were associated with poor response to immunotherapy. Moreover, the SII ≤ 863.3 group had lower rates of decreased consciousness levels, admission to the intensive care unit, and mechanical ventilation; lower cerebrospinal fluid pressure, blood neutrophil count, and platelet count; and higher blood lymphocyte count and GCS scores. The SII was associated with worsened disease severity and poor response to treatment after 30 days of the initially diagnosed AE, and patients with an SII > 863.3 were more likely to have poor response to immunotherapy.

Recent studies demonstrated cutaneous phosphorylated α synuclein (p-syn) deposition in idiopathic and some monogenetic Parkinson disease (PD) patients, suggesting synucleinopathy identical to that in the brain. Although the LRRK2 Gly2385Arg (G2385R) variant is a common PD risk factor in the Chinese population, the pathogenesis of PD with G2385R variant has not been reported. We investigated whether synucleinopathy and small fiber neuropathy (SFN) are associated with the G2385R variant. We performed genotyping in 59 PD patients and 30 healthy controls from the skin biopsy database. The scale of SFN was assessed, as well as bright-field immunohistochemistry against antiprotein gene product 9.5 (PGP9.5) and double-labeling immunofluorescence with anti-PGP9.5 and anti-p-syn. (1) p-syn deposited in the skin nerve fibers of G2385R carrier PD patients, which was a different pattern from noncarriers, without no difference observed between proximal and distal regions; (2) decreased distal intraepidermal nerve fiber density was found in both the G2385R carrier and the noncarrier PD group, and was negatively correlated with composite autonomic symptom score-31 item (COMPASS-31) scores; (3) PD patients with the G2385R variant showed a more peculiar clinical profile than noncarriers with a higher nonmotor symptoms scale, COMPASS-31 score, and levodopa equivalent dose, in addition to an increased prevalence of certain autonomic symptoms or rapid eye movement sleep behavior disorders. Synucleinopathy is related to the LRRK2 G2385R genotype and implies a different pathogenesis in G2385R variant carriers and noncarriers. This study also extended the clinical profiles of PD patients with the G2385R variant.

RATIONALE:Minor ischemic stroke attack has taken a significant part of cerebrovascular disease burden. Benefits of thrombolysis in minor stroke is under debates and the use of urokinase in developing countries needs to be further explored. AIM:TRUST (ThRombolysis of Urokinase for minor STroke) trial was designed to evaluate the efficacy and safety of intravenous urokinase for the treatment of acute minor ischemic stroke. SAMPLE SIZE ESTIMATES:To reach a double-sided type I error rate of 0.05 to test our hypothesis, with β = 0.80, sample size of 1002 subjects were determined after further adjustment to account for up to 5% nonadherence. METHODS AND DESIGN:TRUST trial was developed with PROBE design, as a multicenter, randomized, open label, single-blind clinical trial with the stage of phase 3b. STUDY OUTCOMES:The proportion of patients retaining full ability of independent living, which is defined as patients scoring 0-1 on modified Rankin Scale score at 90 days. DISCUSSION:TRUST trial may potentially provide promising and affordable thrombolysis for acute minor ischemic stroke in the developing parts of the world.

Background: Recent studies have found deposition of phosphorylated α-synuclein (p-syn) in Parkinson disease (PD) patients' skin, indicating p-syn may be a potential biomarker of PD. However, the sensitivity of the p-syn detection varied largely from 5. 3 to 100%, this influenced the clinical use of this detection method to some extent. Objective: This study aimed to optimize the skin biopsy method for detecting p-syn deposition in patients with PD. Methods: Ninety PD patients and 30 healthy controls underwent skin biopsies at 2-3 of the following sites: the distal leg, thigh, cervical region, or forearm. Skin biopsy samples were cut to 50- and 15-μm thickness sections. Deposition of p-syn were detected by using double immunofluorescence labeling of protein gene production 9.5 (PGP9.5) /p-syn. Statistical data analysis was performed using SPSS 25.0 software. Results: Deposition of p-syn were found in 75/90 PD patients but not in healthy controls (p < 0.001). The positive deposition rate of p-syn in the single cervical site was significantly higher than that in the distal leg, thigh, and forearm site. Two samples from the cervical region had a higher p-syn positive rate compared to single cervical site (90.5 vs. 66.7%, p = 0.037). There was no significant difference between the p-syn positive rate of samples from the distal leg/cervical sites and 2 samples from cervical region (80 vs. 90.5%, p = 0.261). Next, the p-syn positive deposition rate of 2-biopsy samples including distal leg/cervical sites and double samples in the cervical site were comparable to the 3-biopsy samples. The 50-μm section had a significantly higher p-syn positive rate than the 15-μm section (p = 0.049). Conclusions: Two biopsy sites (cervical/distal leg) or 2 samples from the cervical site were considered to be priority biopsy sites for detecting p-syn in PD patients. Thick sections may provide a higher p-syn positive rate than thin sections for skin biopsies. These findings provide an optimized p-syn detection method, indicate the valuable pathology biomarker of PD and will promote the clinical use of skin biopsy in the future.

Simple and pure synthetic coating substrates are needed to overcome the disadvantages of traditional coating products like animal derived Matrigel in stem cell research. Since integrins are of great importance in cell adhesion and cell-ECM communication, in this study, a commercially available integrin array established by synthetic integrin binding peptides is used to screen coating substrates for iPSCs and NEPs. The results showed that binding peptides of integrin α5β1, αVβ1, αMβ2 and αIIbβ3 supported cell adhesion of iPSCs, while α5β1, αVβ1 and αIIbβ3 binding peptides supported NEPs adhesion. Additionally, integrin α5β1 binding peptide was revealed to support rapid expansion of iPSCs and iPSC-derived NEPs, as well as the process of NEPs generation, with equal efficiency as Matrigel. In this work, we demonstrated that by supporting stem cell growth in an integrin dependent manner, the integrin array and coating system has the potential to develop more precise and efficient systems in neurological disease modeling.

Spinocerebellar ataxia type 3 (SCA3) is a dominantly inherited neurodegenerative disease caused by a CAG repeat expansion in the region of the ATXN3 gene. The main feature of SCA3 is progressive ataxia, which affects balance, gait, and speech. Urine cells (UCs) of a SCA3 patient were successfully translated to induced pluripotent stem cells (iPSCs) by using the Sendai virus delivery system. ZZUi004-A cell line may provide a robust platform for further study of SCA3 pathogenesis as well as drug testing and gene therapy research.

Non-spherical particles of different shapes have unique properties potentially beneficial in self-assembly, biosensing, therapeutic delivery and optical applications. Forming particles with precisely controlled physical and chemical characteristics, particularly using bioinspired or bio-derived materials can open up applications inaccessible to synthetic polymers. Here, a high throughput fabrication process of different shapes of protein based particles at high resolution using photolithography is demonstrated. In contrast to synthetic polymers, the particles shown herein are comprised of the two silk proteins fibroin and sericin. The demonstrated technique of silk protein lithography allows fabrication of monodisperse biopolymer particles with precise geometries ranging from a few to hundreds of microns. Large numbers of particles of controllable aspect ratios can be easily formed, collected and mixed. The particles themselves are mechanically robust and biocompatible, but can be proteolytically degraded over a period of weeks. Owing to the facile fabrication technique that uses benign solvents, bioactive molecules can be encapsulated within these protein matrices. By control of shape, size, thickness and surface properties, particles that may be harvested for optics, delivery or presentation of biologically functional agents, among other applications. (C) 2016 Elsevier Ltd. All rights reserved.

MiR-198 is involved in tumorigenesis, migration, invasion, and metastasis of various malignant cancers. However, the exact expression levels of miR-198 and the molecular mechanism underlying its role in lung adenocarcinoma require further exploration. In this study, quantitative real-time PCR was applied to study miR-198 and serine hydroxymethyltransferase 1 (SHMT1) expression in 47 paired lung adenocarcinoma tissues and adjacent nontumor lung tissues. Clinicopathological characters were analyzed. Pearson’s correlation analysis was used to detect the relationship between miR-198 and SHMT1 expression. The function of miR-198 was explored by measuring cell proliferation, cell apoptosis, and the cell-cycle in vitro and in vivo. The target gene of miR-198 was certified using dual luciferase report assay. We found that in lung adenocarcinoma, miR-198 was significantly downregulated and SHMT1 was inversely upregulated. A strong negative correlation was noticed between miR-198 and SHMT1 expression. Further analysis revealed that miR-198 expression was associated with TNM stage and lymph node metastasis. Upregulated miR-198 could inhibit cell proliferation, enhance cell apoptosis, and lead to cell-cycle arrest in lung adenocarcinoma, which showed a more effective alteration than SHMT1 siRNA. Moreover, we identified SHMT1 as a target gene of miR-198. In conclusion, miR-198 suppressed proliferation of lung adenocarcinoma cells both in vitro and in vivo by directly targeting SHMT1. miR-198 may be a potential therapeutic target for lung adenocarcinoma in the near future.

MiR-198 is involved in tumorigenesis, migration, invasion, and metastasis of various malignant cancers. However, the exact expression levels of miR-198 and the molecular mechanism underlying its role in lung adenocarcinoma require further exploration. In this study, quantitative real-time PCR was applied to study miR-198 and serine hydroxymethyltransferase 1 ( SHMT1 ) expression in 47 paired lung adenocarcinoma tissues and adjacent nontumor lung tissues. Clinicopathological characters were analyzed. Pearson’s correlation analysis was used to detect the relationship between miR-198 and SHMT1 expression. The function of miR-198 was explored by measuring cell proliferation, cell apoptosis, and the cell-cycle in vitro and in vivo. The target gene of miR-198 was certified using dual luciferase report assay. We found that in lung adenocarcinoma, miR-198 was significantly downregulated and SHMT1 was inversely upregulated. A strong negative correlation was noticed between miR-198 and SHMT1 expression. Further analysis revealed that miR-198 expression was associated with TNM stage and lymph node metastasis. Upregulated miR-198 could inhibit cell proliferation, enhance cell apoptosis, and lead to cell-cycle arrest in lung adenocarcinoma, which showed a more effective alteration than SHMT1 siRNA. Moreover, we identified SHMT1 as a target gene of miR-198. In conclusion, miR-198 suppressed proliferation of lung adenocarcinoma cells both in vitro and in vivo by directly targeting SHMT1 . miR-198 may be a potential therapeutic target for lung adenocarcinoma in the near future.

Cerebral hemorrhage (ICH) is a serious stroke subtype, currently lacking effective treatment. Recent research has shown that CD4(+)CD25(+)FOXP3(+) regulatory T cells (Tregs) play a key role in the immune response of ischemic stroke. However, Tregs in human hemorrhagic stroke are poorly investigated. In this study, a total of 90 ICH patients and 60 healthy controls were recruited. The frequency of circulating Tregs, plasma levels of TGF-β and IL-10, and the severity of neural dysfunction in ICH patients were investigated at different time points post ICH. We found that the peripheral frequency of Tregs in ICH patients was significantly increased, accompanied by boosted activated T cells. Importantly, the elevation of circulating Tregs in patients with severe dysfunction was much higher than that in less-severe patients, suggesting that disease severity affects circulating Tregs to exert regulatory function. Furthermore, both TGF-β and IL-10 that are related to the function of Tregs, were also increased in the peripheral blood of ICH patients. Our results demonstrate that Tregs-mediated immune imbalance might affect the development and severity of ICH, and suggest that Tregs may be used as tools and targets of cellular immunotherapy to effectively treat acute hemorrhagic stroke.

Background: Progressive pseudorheumatoid dysplasia (PPD) is an extremely rare autosomal recessive genetic disease caused by mutation of the Wnt1-inducible signaling pathway protein 3 (WISP3) gene. Here, we characterize the clinical manifestations and features of PPD and screen for WISP3 mutations.Materials and methods: We performed genetic testing for PPD in a Chinese family, after investigating the clinical particulars and family history, in addition to 200 healthy individuals, who served as the controls for this study. All 5 exons and the exon-intron boundaries of the WISP3 gene were amplified by polymerase chain reaction (PCR) and sequenced directly.Results: We identified a missense mutation (c.667T>G, p.C223G) in the maternal allele and a nonsense mutation (c.756C>A, p.C252X) in the paternal allele in the two affected individuals. To our knowledge, the mutation c.756C>A has not been reported previously. In these patients, there was a specific period when their condition markedly improved after having been very serious. Moreover, severe compression of lumbar spinal cord led to conspicuous spinal disorders in the proband.Conclusions: Our study suggests that novel C223G and C252X mutations in exon 4 of the WISP3 gene are responsible for PPD in Chinese patients. Furthermore, we report certain unique phenotypic characteristics in our patients. (C) 2015 Elsevier B.V. All rights reserved.

The element of oriS as a necessary origin sequence for Herpes Simplex Virus type 1derived amplicon vector replicating consists of 45 bp oriS core region and flanking sequences. The flanking sequences of oriS located between gene US12/US1 and RS1 in HSV-1 genome were previously demonstrated to enhance oriS-dependent DNA replication, but the direct evidence of different oriS-flanking sequences in the replication of amplicon virions is still lacked. In this study, we performed a functional analysis of OriS-flanking sequences in replication of HSV-1 based amplicon virions by comparing the yields of virions packaged from amplicon vectors with various deletions of oriS-flanking sequences. The results showed that yields of amplicon virions were significantly reduced by deleting a AgeI/RsrII digested 181bp- fragment between the locus of 333bp-152bp upstream the oriS core region and a NcoI/StyI digested 314bp- fragment between the locus of 23bp-337bp downstream the oriS core region. Furthermore, deletion of a 87bp NcoI/SphI fragment at the locus of 250bp-337bp downstream oriS core region led to a dramatically decrease. Our study further demonstrated that oriS-flanking sequences played an important enhancement role in the amplicon vector replication evidenced at virion level.