Divergent electroencephalogram resting-state functional network alterations in subgroups of autism spectrum disorder: a symptom-based clustering analysis.

Autism spectrum disorder (ASD) is characterized by etiological and phenotypic heterogeneity. Despite efforts to categorize ASD into subtypes, research on specific functional connectivity changes within ASD subgroups based on clinical presentations is limited. This study proposed a symptom-based clustering approach to identify subgroups of ASD based on multiple clinical rating scales and investigate their distinct Electroencephalogram (EEG) functional connectivity patterns. Eyes-opened resting-state EEG data were collected from 72 children with ASD and 63 typically developing (TD) children. A data-driven clustering approach based on Social Responsiveness Scales-Second Edition and Vinland-3 scores was used to identify subgroups. EEG functional connectivity and topological characteristics in four frequency bands were assessed. Two subgroups were identified: mild ASD (mASD, n = 37) and severe ASD (sASD, n = 35). Compared to TD, mASD showed increased functional connectivity in the beta band, while sASD exhibited decreased connectivity in the alpha band. Significant between-group differences in global and regional topological abnormalities were found in both alpha and beta bands. The proposed symptom-based clustering approach revealed the divergent functional connectivity patterns in the ASD subgroups that was not observed in typical ASD studies. Our study thus provides a new perspective to address the heterogeneity in ASD research.

Inhibition of NKCC1 Ameliorates Anxiety and Autistic Behaviors Induced by Maternal Immune Activation in Mice.

Autism spectrum disorder (ASD) is thought to result from susceptibility genotypes and environmental risk factors. The offspring of women who experience pregnancy infection have an increased risk for autism. Maternal immune activation (MIA) in pregnant animals produces offspring with autistic behaviors, making MIA a useful model for autism. However, how MIA causes autistic behaviors in offspring is not fully understood. Here, we show that NKCC1 is critical for mediating autistic behaviors in MIA offspring. We confirmed that MIA induced by poly(I:C) infection during pregnancy leads to autistic behaviors in offspring. We further demonstrated that MIA offspring showed significant microglia activation, excessive dendritic spines, and narrow postsynaptic density (PSD) in their prefrontal cortex (PFC). Then, we discovered that these abnormalities may be caused by overexpression of NKCC1 in MIA offspring's PFCs. Finally, we ameliorated the autistic behaviors using PFC microinjection of NKCC1 inhibitor bumetanide (BTN) in MIA offspring. Our findings may shed new light on the pathological mechanisms for autism caused by pregnancy infection.

Effects of the low Fowler's sleep position and methazolamide treatment on sleep bruxism: A randomized controlled trial.

Intracranial pressure is one of the determinants of sympathetic activities, and sleep bruxism is associated with increased sympathetic activities. This study aimed to investigate effects of the low Fowler's sleep position and methazolamide treatment on the occurrence of rhythmic masticatory muscle activities/sleep bruxism episodes in patients with sleep bruxism in a randomized controlled trial. Polysomnographic recordings were performed on the patients with sleep bruxism sleeping in the low Fowler's (15°-30°) or supine position (n = 11), and with methazolamide or placebo treatment (100 mg, 3-4 hr before bedtime, P.O., n = 9), and changes in sleep variables and heart rate variance during sleep in the low Fowler's position or with methazolamide treatment were determined. Sleep bruxism index, number of masseter muscle electromyographic bursts per hour of sleep, ratio of rhythmic masticatory muscle activities/sleep bruxism duration to the total sleep duration, index of total limb movements, index of limb movements with rhythmic masticatory muscle activities, and number of sleep bruxism clusters per hour of sleep in the low Fowler's position and after methazolamide intake were significantly smaller (p < 0.05-0.001) than those in the supine position and after placebo intake, respectively. The low-frequency heart rate variance powers during non-rapid eye movement sleep stage 2 (N2) in the low Fowler's position and with methazolamide treatment were significantly lower (p < 0.05) than those during sleep in the supine position and with placebo treatment, respectively. In conclusion, sleep in the low Fowler's position and methazolamide treatment were associated with significant decreases in the occurrence of rhythmic masticatory muscle activities/sleep bruxism episodes, which might be due to a reduction in intracranial pressure and sympathetic activities mainly during non-rapid eye movement sleep stage 2.

Study on the Consistency Between Automatic Measurement Based on Convolutional Neural Network Technology and Manual Visual Evaluation in Intracavitary Ultrasonic Cine of Anterior Pelvic.

OBJECTIVES: This study was to evaluate the application of automatic measurement based on convolutional neural network (CNN) technology in intracavitary ultrasound cine of anterior pelvic. METHODS: A total of 500 patients who underwent pelvic floor ultrasound examination at Peking University Shenzhen Hospital from July 2021 to February 2022 were retrospectively retrieved by the picture archiving and communication system (PACS) system, and 300 cases were used as a training set. The training set was labeled by three experienced ultrasound physicians to train CNN models and develop an automatic measurement software. The remaining 200 cases were used as a test set. Automatic measurement software identified relevant anatomical structures frame by frame and determined the two frames with the greatest difference, calculated the bladder neck descent (BND), urethral rotation angle (URA), and retrovesical angle (RA). Meanwhile, two experienced ultrasound physicians evaluated the resting frame and the maximum Valsalva frame on the cines by manual visual evaluation, labeled the anatomical structures in the corresponding frame, such as the inferoposterior margin of pubic symphysis, the mid-axis of pubic symphysis, bladder contour, and urethra in the front, and calculated BND, URA, and RA. Considering that the residual urine volume (RUV) in the bladder may affect the results, enrolled patients were grouped according to the RUV (10-50 mL, 50-100 mL, and >100 mL). The consistency of the results by automatic measurement and manual visual evaluation was evaluated using the intraclass correlation coefficient (ICC) and the Bland-Altman graph. RESULTS: Of the 200 cases in the test set, 120 cases were successfully identified by the CNN automatic software with a 60% recognition rate. In the case of successful identification, the ICC of manual visual evaluation measurement and automatic measurement was 0.936 (BND), 0.911 (URA), 0.756 (RA in rest), and 0.877 (RA at maximum Valsalva), respectively. In addition, the RUV had a negligible effect on the consistency. The Bland-Altman plot shows the proportion of samples outside the limit was below 5%. CONCLUSIONS: CNN-based automatic measurement software exhibited high reliability in anterior pelvic measurement, which results in a significantly enhanced measurement efficiency.

The Prevalence and Risk Factors of Blepharoptosis in an Elderly Asian Population.

BACKGROUND: Age-related blepharoptosis, or ptosis, affects vision and appearance. Associations with age, gender, BMI, and diabetes have been explored, but the link to blood lipids remains unclear. The impact on refraction also lacks consensus. This study addresses gaps by investigating ptosis prevalence and factors in a representative Chinese population, aiming for a comprehensive understanding. METHODS: A cross-sectional study was conducted among individuals aged 50 and above who were willing to participate in comprehensive systemic check-ups, behavioral questionnaires, and ophthalmic examinations at Yaoxi Community Health Center in Wenzhou City, Zhejiang Province. RESULTS: The prevalence of blepharoptosis among the elderly participants at this health center was 27.16%. Individuals with blepharoptosis tended to be older, male, exhibited slightly higher body mass index, wider waist circumference, engaged in lower exercise frequency, and had a higher prevalence of hypertension, diabetes, and with-the-rule astigmatism compared to their counterparts without these conditions. Adjusting for all other confounding variables, older age, being male, higher fasting plasma glucose (FPG), and lower exercise frequency displayed statistically significant relationships with blepharoptosis. After examining the distribution of blepharoptosis degrees within relevant factor subgroups, we noted a higher prevalence of severe ptosis in subgroups associated with older age, male gender, higher FPG, and against-the-rule astigmatism. CONCLUSION: The notable associations with age, gender, FPG, and exercise level suggest a multifactorial etiology for blepharoptosis. The observed link between with-the-rule astigmatism and blepharoptosis implies a potential contributory role in the refractive aspect of blepharoptosis. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Feasibility of a kinect-based system in assessing physical function of the elderly for home-based care.

BACKGROUND: With concerns about accurate diagnosis through telehealth, the Kinect sensor offers a reliable solution for movement analysis. However, there is a lack of practical research investigating the suitability of a Kinect-based system as a functional fitness assessment tool in homecare settings. Hence, the objective of this study was to evaluate the feasibility of using a Kinect-based system to assess physical function changes in the elderly. METHODS: The study consisted of two phases. Phase one involved 35 young healthy adults, evaluating the reliability and validity of a Kinect-based fitness evaluation compared to traditional physical examination using the intraclass correlation coefficient (ICC). Phase two involved 665 elderly subjects, examining the correlation between the Kinect-based fitness evaluation and physical examination through Pearson's correlation coefficients. A Kinect sensor (Microsoft Xbox One Kinect V2) with customized software was employed to capture and compute the movement of joint centers. Both groups performed seven functional assessments simultaneously monitored by a physical therapist and the Kinect system. System usability and user satisfaction were assessed using the System Usability Scale (SUS) and Questionnaire for User Interface Satisfaction (QUIS), respectively. RESULTS: Kinect-based system showed overall moderate to excellent within-day reliability (ICC = 0.633-1.0) and between-day reliability (ICC = 0.686-1.0). The overall agreement between the two devices was highly correlated (r ≧ 0.7) for all functional assessment tests in young healthy adults. The Kinect-based system also showed a high correlation with physical examination for the functional assessments (r = 0.858-0.988) except functional reach (r = 0.484) and walking speed(r = 0.493). The users' satisfaction with the system was excellent (SUS score = 84.4 ± 18.5; QUIS score = 6.5-6.7). CONCLUSIONS: The reliability and validity of Kinect for assessing functional performance are generally favorable. Nonetheless, caution is advised when employing Kinect for tasks involving depth changes, such as functional reach and walking speed tests for their moderate validity. However, Kinect's fundamental motion detection capabilities demonstrate its potential for future applications in telerehabilitation in different healthcare settings.

Effect of Flowering Stages on the Content of Active Ingredients and Antioxidant Capability of Bletilla striata Flowers.

Bletilla striata (Thunb.) Reichb.f. is a perennial herb with abundant active ingredients. Previous research mainly focused on its tubers, however, the study on flowers, especially the variation of active ingredient contents at different flowering stages, was rarely seen. This study analyzed the total phenols, flavonoids, polysaccharides, anthocyanins, and cyanidin-3-O-glucoside content of B. striata flowers which were in cultivated in Herb Garden of Zhejiang A&F University and collected in May, 2019, in order to investigate the changes in active ingredients and antioxidant capacity among different flowering stages (bud, initial, and full bloom). Changes in radical scavenging capability of DPPH (1,1-Diphenyl-2-picrylhydrazyl radical), ABTS (2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonate)), and hydroxy were analyzed. Significant differences in active ingredient content of flowers were detected among different flowering stages. The total phenolic content increased continuously during the entire flowering stage. The contents of total flavonoid, total polysaccharide, and cyanidin-3-O-glucoside reached peaks at the initial blooming stage and then fell as the flowering process continued. The antioxidant activity in initial stage was the highest than in any other flowering stages. Therefore, we conclude that the initial blooming stage is the best harvesting stage of B. striata flowers. This study provides a robust basis for the harvest and utilization of B. striata flowers in food, medical, and cosmetic industries.

RNA-Seq Analysis Reveals an Essential Role of the cGMP-PKG-MAPK Pathways in Retinal Degeneration Caused by Cep250 Deficiency.

Usher syndrome (USH) is characterised by degenerative vision loss known as retinitis pigmentosa (RP), sensorineural hearing loss, and vestibular dysfunction. RP can cause degeneration and the loss of rod and cone photoreceptors, leading to structural and functional changes in the retina. Cep250 is a candidate gene for atypical Usher syndrome, and this study describes the development of a Cep250 KO mouse model to investigate its pathogenesis. OCT and ERG were applied in Cep250 and WT mice at P90 and P180 to access the general structure and function of the retina. After recording the ERG responses and OCT images at P90 and P180, the cone and rod photoreceptors were visualised using an immunofluorescent stain. TUNEL assays were applied to observe the apoptosis in Cep250 and WT mice retinas. The total RNA was extracted from the retinas and executed for RNA sequencing at P90. Compared with WT mice, the thickness of the ONL, IS/OS, and whole retina of Cep250 mice was significantly reduced. The a-wave and b-wave amplitude of Cep250 mice in scotopic and photopic ERG were lower, especially the a-wave. According to the immunostaining and TUNEL stain results, the photoreceptors in the Cep250 retinas were also reduced. An RNA-seq analysis showed that 149 genes were upregulated and another 149 genes were downregulated in Cep250 KO retinas compared with WT mice retinas. A KEGG enrichment analysis indicated that cGMP-PKG signalling pathways, MAPK signalling pathways, edn2-fgf2 axis pathways, and thyroid hormone synthesis were upregulated, whereas protein processing in the endoplasmic reticulum was downregulated in Cep250 KO eyes. Cep250 KO mice experience a late-stage retinal degeneration that manifests as the atypical USH phenotype. The dysregulation of the cGMP-PKG-MAPK pathways may contribute to the pathogenesis of cilia-related retinal degeneration.

2019 Global NAFLD Prevalence: A Systematic Review and Meta-analysis.

BACKGROUND & AIMS: The increasing rates of obesity and type 2 diabetes mellitus may lead to increased prevalence of nonalcoholic fatty liver disease (NAFLD). We aimed to determine the current and recent trends on the global and regional prevalence of NAFLD. METHODS: Systematic search from inception to March 26, 2020 was performed without language restrictions. Two authors independently performed screening and data extraction. We performed meta-regression to determine trends in NAFLD prevalence. RESULTS: We identified 17,244 articles from literature search and included 245 eligible studies involving 5,399,254 individuals. The pooled global prevalence of NAFLD was 29.8% (95% confidence interval [CI], 28.6%-31.1%); of these, 82.5% of included articles used ultrasound to diagnose NAFLD, with prevalence of 30.6% (95% CI, 29.2%-32.0%). South America (3 studies, 5716 individuals) and North America (4 studies, 18,236 individuals) had the highest NAFLD prevalence at 35.7% (95% CI, 34.0%-37.5%) and 35.3% (95% CI, 25.4%-45.9%), respectively. From 1991 to 2019, trend analysis showed NAFLD increased from 21.9% to 37.3% (yearly increase of 0.7%, P < .0001), with South America showing the most rapid change of 2.7% per year, followed by Europe at 1.1%. CONCLUSIONS: Despite regional variation, the global prevalence of NAFLD is increasing overall. Policy makers must work toward reversing the current trends by increasing awareness of NAFLD and promoting healthy lifestyle environments.

Effects of dietary dandelion extract on the growth performance, serum biochemical parameters, liver histology, and immune and apoptosis-related genes expression of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) at different feeding period.

The study explored on the effects of dietary 0.4% dandelion extract on the growth performance, serum biochemical parameters, liver histology and the expression levels of immune and apoptosis-related genes in the head kidney and spleen of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) at different feeding period. The results showed that the weight gain rate (WGR) of the hybrid grouper were significantly increased at the second and fourth weeks (P < 0.05), but there was no significant difference in WGR at the eighth week (P > 0.05). Compared with the control group, dietary dandelion extracts supplementation improve lipid metabolism, reduce lipid accumulation in liver and maintain normal liver histology at the second and fourth weeks. At the end of the second week, the relative expression levels of antioxidant related genes (MnSOD, GPX and GR) in the head kidney of hybrid grouper fed with dandelion extract increased significantly; at the end of week 4 and week 8, the relative expression levels of antioxidant related genes other than MnSOD did not change significantly. However, in the spleen of hybrid grouper, the expression of these antioxidant genes showed the opposite trend. At the end of the eighth week, dietary dandelion extract supplementation significantly increased the expression of inflammatory response related genes in head kidney of hybrid grouper, but showed the opposite trend in spleen. In conclusion, the short-term (2 or 4 weeks) application of 0.4% dandelion extract in feed had the effects of growth improvement, liver protection and immune stimulation on hybrid grouper due to its antioxidant and anti-inflammatory activities. The beneficial effect of dandelion extract on hybrid grouper was time-dependent, and its action time on different immune organs of hybrid grouper was not synchronous.

Characteristics of the optic disc in young people with high myopia.

PURPOSE: This study aimed to investigate the characteristics of the optic disc in adolescents and young adults with high myopia by applying optical coherence tomography angiography. METHODS: A total of 112 patients with high myopia (spherical equivalent refraction (SER) ≤ -6.00 D) aged 12 to 30 years old were enrolled in this cross-sectional study. Parapapillary atrophy (PPA) and ovality index from scanning laser ophthalmoscopy images and the degree of optic disc tilt from the optic nerve head (ONH) OCT B-scans were analysed using ImageJ and MATLAB software. Peripapillary retinal nerve fibre layer thickness (pRNFLT) and radial peripapillary capillary vessel density (RPC VD) around the optic disc were obtained from the images of the optic disc angiography scan. RESULTS: In young high myopia patients, the PPA area was positively correlated with age, axial length (AL) and pRNFLT (all p < 0.05) and negatively correlated with SER (r = -0.222, p = 0.020). The degree of the optic disc tilt was associated with increasing AL and pRNFLT (all p < 0.05). The disc area was positively correlated with AL, pRNFLT, and RPC VD (all p < 0.05). In the multivariate regression analysis, PPA area was independently associated with the degree of optic disc tilt and disc area. The degree of optic disc tilt was affected by AL and PPA area while the change of disc area was influenced by PPA area and pRNFLT (all p < 0.05). CONCLUSION: In young patients with high myopia, PPA area, the degree of optic disc tilt and disc area increased with AL and pRNFLT, while decreased with SER. The association between these factors was slightly different in the adolescent and young adult groups. The degree of the optic disc tilt was more associated with AL and SER in the adolescent group while disc area showed more correlated with AL and SER in the young adult group.

The effects of dietary compound plant extracts on growth performance, liver and intestine health, and immune related genes expression in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀).

The study explored on the effect of dietary compound plant extract supplementation on the growth performance, serum biochemical indicators, liver and intestinal morphological and gene expression levels in the head kidney and spleen of the hybrid grouper (Epinephelus lanceolatusâ™‚× Epinephelus fuscoguttatus♀). The compound plant extracts (BDG) was a mixture of Bupleurum edulis extract, dandelion extract and Ginkgo biloba extract in a ratio of 1:4:1. Basal diets supplemented with BDG at 0, 0.75, 1.5, 3 and 6 g/kg were fed hybrid grouper for 8 weeks. The results showed that dietary 0.75 and 1.5 g/kg BDG supplementation could significantly increase the WGR and SGR of hybrid grouper (P < 0.05). And dietary 0.75 g/kg BDG could also significantly decrease serum aspartate aminotransferase, glucose and lactate dehydrogenase in hybrid grouper (P < 0.05). Dietary BGD supplementation protected the integrity of liver and intestinal morphological structure, reduced the accumulation of liver fat. Dietary BDG supplementation might enhance the immunity of hybrid grouper by regulating the expression of antioxidant and inflammation-related genes in head kidney and spleen of hybrid grouper. Our study demonstrated that the growth promoting effect of Bupleurum extract, dandelion extract and Ginkgo biloba extract in the ratio of 1:4:1 as a compound feed additive was better than any of them as a feed additive alone, and the dosage was less. The optimal additive dosage of BDG was 0.75 g/kg in hybrid grouper diets.

Associations between optic disc characteristics and macular choroidal microvasculature in young patients with high myopia.

BACKGROUND: This study aimed to examine changes to optic disc characteristics and macular choroidal microvasculature, and their relationships in young patients with high myopia (HM). METHODS: A total of 90 patients were enrolled in this cross-sectional study. Based on their refractive power, the patients were divided into three groups: 27 in the control group, 34 in the HM group and 29 in the extremely high myopia group. Images of each patient's macula and optic disc were taken by ocular coherence tomography angiography. The macular choroidal and retinal thickness, capillary vessel density and capillary flow area were measured using Matlab software. Parapapillary atrophy (PPA) and the ovality index (OI) obtained from the scanning laser ophthalmoscopy images and the degree of optic disc tilt obtained from the optic nerve head ocular coherence tomography B-scans were analysed by Image J and Matlab software. RESULTS: The PPA area, OI and degree of optic disc tilt were significantly different among the three groups (all p ≤ 0.001). The macular choroidal thickness and microvasculature were significantly different among the three groups (all p < 0.05). Macular choroidal thickness was significantly correlated with PPA area and the degree of optic disc tilt (r = -0.331, p = 0.003; r = -0.394, p = 0.001, respectively). Macular choroidal capillary vessel density and choriocapillaris flow area were associated with PPA area (r = -0.251, p = 0.047; r = -0.326, p = 0.009, respectively). CONCLUSIONS: PPA area, OI and the degree of optic disc tilt were increased in patients with HM, and these changes were correlated with macular choroidal thickness and choroidal microvasculature.

Experiences with cancer survey in China.

BACKGROUND: China accounts for approximately 27% of global cancer deaths. However, to the authors' knowledge, the lasting effects of cancer and cancer treatments on patients have not been investigated in China. The authors developed a questionnaire, the China Survey of Experiences with Cancer, for Chinese cancer survivors. This article introduces the study design and domains covered in the questionnaire. METHODS: The Cancer Survivorship Supplement of the Medical Expenditure Panel Survey (MEPS) was used as a reference to develop the questionnaire. The final in-person surveys were conducted in 2015 and 2016. Samples were chosen through multistage sampling. The authors described the characteristics of the study participants and their cancer experiences. RESULTS: At the time of last follow-up, a total of 1166 patients had completed at least 1 component of the survey. Approximately 59% of the cancer survivors were aged ≥60 years. Greater than one-half of the participants had an elementary education level or less (51%) and a yearly family income of <$3174. Chinese cancer survivors were more likely to retire earlier than planned compared with American cancer survivors (37% vs 9%). The majority of Chinese cancer survivors (84%) reported that their work abilities were hindered by their cancer or cancer treatments. Approximately one-half of patients in China had to incur debt because of cancer, whereas <10% of patients in the United States reported having incurred debt. CONCLUSIONS: The survey provides information regarding the burden of cancer in China that to the authors' knowledge currently is unavailable from other sources, including medical care use, financial impacts, employment patterns, and life experience after a cancer diagnosis for survivors and their families.

THREE-DIMENSIONAL CHARACTERISTICS OF FOUR MACULAR INTRARETINAL LAYER THICKNESSES IN SYMPTOMATIC AND ASYMPTOMATIC CARRIERS OF G11778A MUTATION WITH LEBER'S HEREDITARY OPTIC NEUROPATHY.

PURPOSE: To characterize by spectral domain optical coherence tomography the three-dimensional thicknesses of four macular intraretinal layers in symptomatic and asymptomatic carriers of G11778A mutation with Leber's hereditary optic neuropathy. METHODS: Twenty-five eyes (7 symptomatic eyes and 18 asymptomatic eyes) of patients with Leber's hereditary optic neuropathy from one Chinese family and 16 normal eyes were enrolled. Macular radial scans by spectral domain optical coherence tomography and custom software produced intraretinal three-dimensional thickness maps. The macula was divided into nine regions, and each region included four intraretinal layers: nerve fiber layer, ganglion cell layer and inner plexiform layer, inner nuclear layer and outer plexiform layer, and the outer retinal layer. RESULTS: Nerve fiber layer in the symptomatic eyes was significantly thinner than in normal eyes for most of the macular regions; however in the asymptomatic eyes, it was increased in three regions. Ganglion cell layer and inner plexiform layers in all regions of symptomatic eyes were significant thinner than in asymptomatic eyes and controls. Inner nuclear layer and outer plexiform layers in six regions of symptomatic and asymptomatic eyes were significantly thicker than in controls. The outer retinal layer of asymptomatic eyes was thicker than in most control regions. CONCLUSION: Intraretinal thickness changes in asymptomatic patients could be prodromal events that indicate the imminent conversion to symptomatic patients with Leber's hereditary optic neuropathy.

Immunogenicity of oral vaccination with Lactococcus lactis derived vaccine candidate antigen (UreB) of Helicobacter pylori fused with the human interleukin 2 as adjuvant.

Helicobacter pylori (H. pylori) infection remains a significant global public health problem. Vaccine, especially edible vaccine, is considered to be effective in the management of H. pylori infections. By using recombinant technology, Lactococcus lactis (L. lactis) could serve as an antigen-delivering vehicle for the development of edible vaccine. The aim of this study was to produce edible UreB (urease B) vaccine derived from L. lactis against H. pylori. The UreB subunit is the most effective and common immunogen of all strains of H. pylori. The UreB was produced as a chimeric protein fused with IL-2 (human interleukin 2) as the mucosal adjuvant. Mucosal immunization of mice with recombinant L. lactis NZ9000 containing the UreB-IL-2 protein elicited more anti-UreB antibody that specifically bounded to the purified bacterial UreB protein and more cytokines such as IFN-γ, IL-4, and IL-17, and had a lower H. pylori burden and urease activity than control mice. These results suggest that the recombinant L. lactis expressing UreB-IL-2 can be potentially used as an edible vaccine for controlling H. pylori infection.

Infantile epileptic spasm syndrome as a new NR2F1 gene phenotype.

INTRODUCTION: NR2F1 pathogenetic variants are associated with the Bosch-Boonstra-Schaaf optic atrophy syndrome (BBSOAS). Recent studies indicate that BBSOAS patients not only have visual impairments but may also have developmental delays, hypotonia, thin corpus callosum and epileptic seizures. However, reports of BBSOAS occurrence along with infantile epileptic spasm syndrome (IESS) are rare. METHODS: Here, we report three cases involving children with IESS and BBSOAS caused by de novo NR2F1 pathogenetic variants and summarize the genotype, clinical characteristics, diagnosis and treatment of them. RESULTS: All three children experienced epileptic spasms and global developmental delays, with brain Magnetic Resonance Imaging (MRI) suggesting abnormalities (thinning of the corpus callosum or widened extracerebral spaces) and two of the children exhibiting abnormal visual evoked potentials. CONCLUSIONS: Our findings indicate that new missense NR2F1 pathogenetic variants may lead to IESS with abnormal visual evoked potentials. Thus, clinicians should be aware of the Bosch-Boonstra-Schaaf optic atrophy syndrome and regular monitoring of the fundus, and the optic nerve is necessary during follow-up.

Replacement of fish meal with cottonseed protein concentrate in Chinese mitten crab (Eriocheir sinensis): Nutrient digestibility, growth performance, free amino acid profile, and expression of genes related to nutrient metabolism.

This study aimed to investigate the application of cottonseed protein concentrate (CPC) in Chinese mitten crabs (Eriocheir sinensis). First, the apparent digestibility coefficient (ADC) of CPC, fish meal and soybean meal were compared in crabs (21.72 ± 0.33 g). The protein ADC of CPC was 90.42%, which was significantly higher than that of soybean meal (83.16%) (P < 0.05). The ADC of Phe, Cys and Glu of CPC were significantly higher than those of fish meal, while the ADC of Ile, Leu, Lys, Met, Thr and Ala of CPC were significantly lower (P < 0.05). Second, we investigated the effects of fish meal substitution by CPC on growth performance, free amino acid profile, and expression of genes related to nutrient metabolism in crabs. Six diets were formulated by replacing 0%, 15%, 30%, 45%, 60% and 75% fish meal with CPC, namely FM, CPC15, CPC30, CPC45, CPC60, and CPC75. A total of 630 crabs (1.68 ± 0.00 g) were randomly divided into 18 tanks (3 tanks per group) and fed 3 times daily for 9 weeks. Results showed that CPC75 group significantly reduced growth performance, feed conversion efficiency, and free Ile, Leu, Lys, Met, and Thr contents in muscle (P < 0.05). The contents of free amino acids (Arg, His, Ile, Leu, Lys, Met, Phe, Thr, Val, Ala, Cys, Glu, Gly, Ser and Tyr) in hepatopancreas decreased linearly with the increase of dietary CPC level (P < 0.05). The substitution of more than 45% fish meal with CPC significantly decreased the concentration of delicious amino acids (Ala, Glu and Gly) in hepatopancreas (P < 0.05), which might adversely affect crab flavor. The expression of genes related to antioxidant capacity, protein transport, TOR pathway and lipid metabolism was significantly downregulated by increasing dietary CPC level (P < 0.05). In conclusion, based on the quadratic regression analysis of FCR and PER, the optimal replacement levels of fish meal with CPC in crab diet containing 35% fish meal were 32.36% and 35.38%, respectively. It is recommended that Ile, Leu and Thr be supplemented in addition to Met and Lys in the application of CPC.

Expanding the clinical phenotype and genetic spectrum of GEMIN5 disorders: Early-infantile developmental and epileptic encephalopathies.

BACKGROUND: Several biallelic truncating and missense variants of the gem nuclear organelle-associated protein 5 (GEMIN5) gene have been reported to cause neurodevelopmental disorders characterized by cerebellar atrophy, intellectual disability, and motor dysfunction. However, the association between biallelic GEMIN5 variants and early-infantile developmental and epileptic encephalopathies (EIDEEs) has not been reported. PURPOSE: This study aimed to expand the phenotypic spectrum of GEMIN5 and explore the correlations between epilepsy and molecular sub-regional locations. METHODS: We performed whole-exome sequencing in two patients with EIDEE with unexplained etiologies. The damaging effects of variants were predicted using multiple in silico tools and modeling. All reported patients with GEMIN5 pathogenic variants and detailed neurological phenotypes were analyzed to evaluate the genotype-phenotype relationship. RESULTS: Novel biallelic GEMIN5 variants were identified in two unrelated female patients with EIDEE, including a frameshift variant (Hg19, chr5:154284147-154284148delCT: NM_015465: c.2551_c.2552delCT: p.(Leu851fs*30)), a nonsense mutation (Hg19, chr5:154299603-154299603delTinsAGA: NM_015465: c.1523delTinsAGA: p.(Leu508*)), and two missense variants (Hg19, chr5:154282663T > A: NM_015465: c.2705T > A: p.(Leu902Gln) and Hg19, chr5:154281002C > G: NM_015465: c.2911C > G: p.(Gln971Glu)), which were inherited from asymptomatic parents and predicted to be damaging or probably damaging using in silico tools. Except p.Leu508*, all these mutations are located in tetratricopeptide repeat (TPR) domain. Our two female patients presented with seizures less than 1 month after birth, followed by clusters of spasms. Brain magnetic resonance imaging suggests dysgenesis of the corpus callosum and cerebellar hypoplasia. Video electroencephalogram showed suppression-bursts. Through a literature review, we found 5 published papers reporting 48 patients with biallelic variants in GEMIN5. Eight of 48 patients have epilepsy, and 5 patients started before 1 year old, which reminds us of the relevance between GEMIN5 variants and EIDEE. Further analysis of the 49 GEMIN5 variants in those 50 patients demonstrated that variants in TPR-like domain or RBS domain were more likely to be associated with epilepsy. CONCLUSIONS: We found novel biallelic variants of GEMIN5 in two individuals with EIDEE and expanded the clinical phenotypes of GEMIN5 variants. It is suggested that the GEMIN5 gene should be added to the EIDEE gene panel to aid in the clinical diagnosis of EIDEE and to help determine patient prognosis.