Effects of green tea powder on production performance, egg quality, and blood biochemical parameters in laying hens.

The paper aimed to evaluate the effects of dietary inclusion of green tea powder (GTP) on laying performance, egg quality, and blood biochemical parameters of laying hens. A total of 240 Jingfen No. 6 laying hens (age, 24 wk) were randomly allocated into 4 groups: control group (CON, basal diet), GTP0.5, GTP0.75, and GTP1.0 (basal diet included 0.5, 0.75, and 1.0% GTP, respectively). Each group has 5 replicates with 12 birds each. The feeding trial lasted 8 wk. The results showed that the hen-day egg production rate in GTP0.5 and GTP 0.75 group was higher than that of GTP1.0 group (P < 0.05), hen-day egg production rate in the GTP1.0 group was lower compared to the CON group (P > 0.05), the feed conversion ratio (FCR) in the GTP0.75 group was lower than that in CON and GTP 1.0 group (P < 0.05) during the entire experimental period. Albumen height and Haugh unit were higher in the GTP0.75 and GTP1.0 group compared to the CON group at d 56 (P < 0.05). At the end of experiment, plasma TG content in the GTP0.75 and GTP1.0 group was lower than that in the CON group (P < 0.05), the T-CH concentration in the GTP0.5 and GTP0.75 group was lower compared to the CON group (P < 0.05), plasma LDL-C and CORT concentrations were decreased by dietary GTP supplementation (P < 0.05), the HDL-C and BUN concentrations in the GTP0.75 and GTP1.0 group were higher than that in the CON group (P < 0.05). The antibody titers of H5N1 in the GTP0.75 and GTP1.0 group, and H7N9 in the GTP1.0 group were lower than that in the CON group (P < 0.05). In conclusion, dietary GTP inclusion could affect laying performance, regulate lipid metabolism, and have no favorable influence on antibody titers of H5N1 and H7N9, herein, dietary 0.5% GTP inclusion is suggested for Jingfen No. 6 laying hens during the peak laying period.

Permanent Shape Reconfiguration and Locally Reversible Actuation of a Carbon Nanotube/Ethylene Vinyl Acetate Copolymer Composite by Constructing a Dynamic Cross-Linked Network.

Given the rapid developments in modern devices, there is an urgent need for shape-memory polymer composites (SMPCs) in soft robots and other fields. However, it remains a challenge to endow SMPCs with both a reconfigurable permanent shape and a locally reversible shape transformation. Herein, a dynamic cross-linked network was facilely constructed in carbon nanotube/ethylene vinyl acetate copolymer (CNT/EVA) composites by designing the molecular structure of EVA. The CNT/EVA composite with 0.05 wt % CNT realized a steady-state temperature of ∼75 °C under 0.11 W/cm2 light intensity, which gave rise to remote actuation behavior. The dynamic cross-linked network along with a wide melting temperature offered opportunities for chemical and physical programming, thus realizing the achievement of the programmable three-dimensional (3D) structure and locally reversible actuation. Specifically, the CNT/EVA composite exhibited a superior permanent shape reconfiguration by activating the dynamic cross-linked network at 140 °C. The composite also showed a high reversible deformation rate of 11.1%. These features endowed the composites with the capability of transformation to 3D structure as well as locally reversible actuation performance. This work provides an attractive guideline for the future design of SMPCs with sophisticated structures and actuation capability.

GABAergic neurons in the ventral lateral geniculate nucleus and intergeniculate leaflet are involved in itch processing in mice.

Itch and pain are two closely related sensations that receiving similar encodings at multiple levels. Accumulated evidences suggest that activation of the ventral lateral geniculate nucleus and intergeniculate leaflet (vLGN/IGL)-to-lateral and ventrolateral periaqueductal gray (l/vlPAG) projections mediates the antinociceptive effects of bright light therapy. Clinical study showed that bright light therapy may ameliorate cholestasis-induced pruritus. However, the underlying mechanism and whether this circuit participates in itch modulation remains unclear. In this study, chloroquine and histamine were utilized to induce acute itch models in mice. Neuronal activities in vLGN/IGL nucleus were evaluated with c-fos immunostaining as well as fiber photometry. Optogenetic manipulations were performed to activate or inhibit GABAergic neurons in the vLGN/IGL nucleus. Our results showed that the expressions of c-fos in vLGN/IGL were significantly increased upon both chloroquine- and histamine-induced acute itch stimuli. GABAergic neurons in vLGN/IGL were activated during histamine and chloroquine-induced scratching. Optogenetic activation of the vLGN/IGL GABAergic neurons exerts antipruritic effect, while inhibiting these neurons exerts pruritic effect. Our results provide evidence that GABAergic neurons in vLGN/IGL nucleus might play a crucial role in modulating itch, which may provide clue for application of bright light as an antipruritic treatment in clinic.

[Clinical application and evaluation of health economics for non-invasive prenatal testing of fetuses in Tianjin].

OBJECTIVE: To assess the clinical efficacy and health economic value of non-invasive prenatal testing (NIPT) for the prenatal screening of common fetal chromosomal aneuploidies. METHODS: 10 612 pregnant women from October 2017 to December 2019 presented at the antenatal screening clinic of the General Hospital of Tianjin Medical University were selected as the study subjects. Results of NIPT and invasive prenatal diagnosis and follow-up outcome for the 10 612 pregnant women were retrospectively analyzed and compared. Meanwhile, NIPT data for two periods were analyzed for assessing the health economic value of NIPT as the second- or first-tier screening strategy for the prenatal diagnosis of fetal trisomies 21, 18 and 13. RESULTS: The NIPT was successful in 10 528 (99.72%) subjects, with the sensitivity for fetal trisomies 21, 18 and 13 being 100%, 92.86% and 100%, and the positive predictive value (PPV) being 89.74%, 61.90% and 44.44%, respectively. The PPV of NIPT for sex chromosome aneuploidies was 34.21%. Except for one false negative case of trisomy 18, the negative predictive value for trisomy 21, trisomy 13 and other chromosomal abnormalities were 100%. For pregnant women with high risk by serological screening, advanced maternal age or abnormal ultrasound soft markers, NIPT has yielded a significantly increased high risk ratio. There was no statistical difference in the PPV of NIPT among pregnant women from each subgroup. NIPT would have higher health economic value as a second-tier screening until 2019, while compared to 2015 ~ 2017, its incremental cost-effectiveness ratio as a first-tier screening had declined clearly. CONCLUSION: The screening efficacy of NIPT for trisomies 21, 18 and 13 for a mixed population is significantly better than conventional serological screening, but it is relatively low for sex chromosomal abnormalities. NIPT can also be recommended for populations with relatively high risks along with detailed pre- and post-test genetic counselling. From the perspective of health economics, except for open neural tube defects, it is possible for NIPT to replace the conventional serological screening in the future as its cost continues to decrease.

Effects of Different-Sized Cages on the Production Performance, Serum Parameters, and Caecal Microbiota Composition of Laying Hens.

The effects of four different-sized cages­huge (HC), large (LC), medium (MC), and small (SC) cages­on the productive performance, serum biochemical indices, and caecal microbiota composition of Roman laying hens were investigated. At 44 weeks of age, a total of 450 hens were selected and allocated to the four groups, with six replicates each. Equal stocking density (0.054 m2 per bird) was maintained among the four groups throughout the experiment, and number of birds/cage changed for each treatment. After 2 weeks of preliminary trial, the formal experiment was performed from 46 to 60 weeks of age. The laying rate and feed conversion ratio (FCR) were determined daily, antibody titres were measured every 3 weeks, and serum biochemical parameters and caecal microbiota composition were analysed at 60 weeks of age. Compared to HC and SC, the higher laying rate and lower FCR in MC and LC indicated positive effects on egg production and feed efficiency, while SC showed the highest body weight gain (p < 0.05). With increasing cage size, the serum triglycerides (TG) and total cholesterol (T-CH) levels were reduced, and serum glutathione peroxidase (GSH-Px) activity improved, where birds raised in HCs had the lowest serum TG and T-CH and the highest GSH-Px activity. Twenty-nine different phyla and 301 different genera were detected in the caecal microbiota of birds in the four groups. Methanobrevibacter was significantly higher in the SC than in the other groups (p < 0.05). Faecalibacterium was most abundant in the MC compared with the other groups (p < 0.05) and was significantly positively correlated with serum GSH-Px concentration (R = 0.214, p = 0.0017). Lactobacillus was significantly less abundant in the LC and MC than in the HC and SC groups (p < 0.05) and was significantly positively correlated with body weight (R = 0.350, p = 0.0009) but negatively correlated with laying rate and FCR. In conclusion, MC were superior to HC and LC in improving feed conversion efficiency and caecal microflora composition compared to the SC. An appropriate increase in cage size is beneficial to laying hen production and health.

Non-photosynthetic chemoautotrophic CO2 assimilation microorganisms carbon fixation efficiency and control factors in deep-sea hydrothermal vent.

Non-photosynthetic chemoautotrophic microorganisms in deep-sea hydrothermal vent can obtain energy by oxidation reducing substances and synthesize CO2 into organic carbon, and the development and utilization of microbial resources in this environment for CO2 fixation under ordinary environmental conditions is of great significance to understand the carbon cycle and microbial carbon fixation in deep-sea hydrothermal vent. In this study, a set of spiral-stirred bioreactor (SSB) was developed to cultivate a group of non-photosynthetic chemoautotrophic CO2 assimilation microorganisms (NPCAM), mainly Sphingomonadaceae (unclassified, the mean of which was 31.16 %), from deep-sea hydrothermal vent sediments, which have the characteristics of halophilic, acid-base and heavy metal resistant. The maximum carbon fixation efficiency (calculated by CO2) was 6.209 mg·CO2/(L·h) after 96 h of incubation in the presence of mixed electron donors (MEDs, 0.46 % NaNO2, 0.50 % Na2S2O3 and 1.25 % Na2S, w/v), mixed inorganic carbon sources (CO2, Na2CO3 and NaHCO3) and aerobic conditions. The detection of NPCAM synthetic organic fraction in SSB system, the study of single bacteria culturability and carbon fixation efficiency, the analysis of CO2 fixation pathway and the development of coupled carbon fixation technology are the prospective works that need to be further developed.

Evaluation of the practical applications of fluorescence in situ hybridization in the prenatal diagnosis of positive noninvasive prenatal screenings.

OBJECTIVE: To investigate the application value and limitations of fluorescence in situ hybridization (FISH) in prenatal diagnosis of positive results for trisomies 13, 18, 21 (T13, T18, T21) and sex chromosome aneuploidies (SCAs) indicated by noninvasive prenatal screening (NIPS). METHODS: Samples from women who underwent prenatal diagnosis for the indication of positive NIPS of T13, T18, T21, and SCAs were collected. Each sample was split into two for both karyotype analysis and FISH analysis. The efficiency and consistency of FISH were assessed for the detection of chromosome abnormalities in the indications of positive NIPS results compared with karyotyping. RESULTS: A total of 649 pregnant women who scored positive for clinical significance of fetal chromosome abnormalities by NIPS were enrolled in our study, including T 13 (6%), T18 (14.3%), T21 (44.7%), SCAs (35.0%). From the following diagnostic test, the positive predictive value (PPV) of NIPS for T13, T18, T21, and SCAs was 17.9, 60.2, 89.3, and 43.6% respectively. FISH analysis was successful in all samples. Compared with karyotyping, the sensitivity and specificity were 98.3 and 100%, respectively. 95.7% (621/649) were fully concordant with karyotyping. 3.2% (21/649) cases were incompletely concordant with the karyotyping, among these cases, the FISH analysis identified all the aneuploidies, but karyotyping analysis provided more information about the chromosomal structure. There were 7 cases (1.1%, 7/649) of anomalies diagnosed by karyotype but missed out by FISH, all of which occurred in cases with the indication of SCAs. If the indications were confined to cases with a positive NIPS of T13, T18, T21, the diagnostic consistency of the two methods almost perfectly agree, and all the aneuploidies were detected by the FISH assay. FISH analysis was highly consistent in determining whether the fetus was euploid or not in the prenatal diagnosis for the patients with positive NIPS results compared with karyotyping (kappa= 0.976, p < .01). CONCLUSION: For the prenatal diagnostic indications of positive NIPS of T13, T18, T21, and SCAs, FISH was equally efficacious in identifying aneuploidies and provided a quick diagnosis to alleviate anxiety. However, the missed risk of FISH analysis for structural chromosomal abnormalities should be taken seriously and fully informed during genetic counseling.

Effects of dietary fermented peony seed dreg on the laying performance, albumen quality, antioxidant capacity, and n-3 PUFA-enriching property of laying hens.

Introduction: It is of great importance to seek agro-industrial byproducts that can serve as unconventional or alternative feedstuffs for poultry, especially those that are substitutive sources of n-3 polyunsaturated fatty acids (PUFAs), as this will contribute to alleviating feed shortage pressure and improving poultry performance and product quality. In this study, the effects of dietary fermented peony seed dreg (FPSD) on the production performance of hens and fatty acid composition of meat and egg yolk were evaluated. Methods: A total of 480 54-week-old Xinyang chickens were divided into 5 groups: control (basal diet); 5% peony seed dreg (PSD); and 5%, 7% and 9% FPSD, and each group consisted of 6 replicates with 16 birds per replicate. Production performances were recorded daily, and egg quality, serum parameters, antibody titers and fatty acid profile in the muscle and yolk were measured at 64 weeks of age. Results and discussion: Egg production, feed conversion ratio and egg albumen quality were improved (p < 0.05) by 5% and 7% FPSD groups compared to 5% PSD and the control diet. Immune status was not influenced by PSD, but antibody titres against H7N9 and ND were improved by FPSD diets during most of the experimental periods. Dietary 5% and 7% FPSD increased (p < 0.05) serum high density lipoprotein concentrations and glutathione peroxidase actives. Total n-3 polyunsaturated fatty acids (PUFA) in meat and yolk increased gradually, while the total saturated fatty acids (SFA) and the n-6/n-3 PUFA ratio decreased gradually with increasing dietary FPSD levels. In conclusion, up to 7% dietary FPSD has the potential use to be utilized as a supplement in Xinyang laying hen diets to produce n-3 PUFA-enriched meat and eggs and positively affect production performance and health status.

Different Non-cage Housing Systems Alter Duodenal and Cecal Microbiota Composition in Shendan Chickens.

Housing systems are among the most important non-genetic factors affecting hen production performance and intestinal microbes. With increased interest in animal welfare, cage-free laying hen housing systems have become common, providing behavioral freedom and health benefits. The present study aimed to compare the effects of plastic net housing system (NRS) and floor litter housing system (LRS) on the composition and function of the duodenal and cecal microbiota in Shendan chicken, one of the most popular laying hen strains in China. The associations between the differential microbiota abundance and production traits and intestinal morphological parameters were determined. Compared with the LRS, the NRS improved the laying rate (p < 0.05) and increased the villus height (VH) of the duodenum (p < 0.05) and the VH-to-crypt depth ratio (VCR) of the cecum (p < 0.05). Alpha diversity analysis showed that LRS chickens had a significantly higher diversity and richness than NRS chickens. Beta diversity analysis demonstrated differences in the microbiota composition based on housing systems. Within the cecum, Proteobacteria and Kiritimatiellaeota were significantly more abundant in the LRS than in the NRS (p < 0.05), while Bacteroidetes were significantly less abundant in the LRS (p < 0.05). Phascolarctobacterium and Ruminococcaceae_UCG-005 were significantly less abundant in the LRS (p < 0.05) compare to the NRS. Within the duodenum, Lactobacillus was significantly less abundant in the LRS (p < 0.05) than in the NRS, while Pseudomonas was significantly more abundant in the LRS (p < 0.05). Cecal Phascolarctobacterium and Ruminococcaceae_UCG-005 were significantly positively correlated with eggshell strength (R = 0.608, p < 0.01) and egg weight (R = 0.526, p < 0.05), respectively. Duodenal Lactobacillus was significantly positively correlated with VH and VCR (R = 0.548 and 0.565, p < 0.05), while Pseudomonas was significantly negatively correlated with the Haugh unit (R = -0.550, p < 0.05). In conclusion, there are differences in the cecal and duodenal microbiota compositions of Shendan laying hens reared in different non-cage housing systems, and the NRS was superior to the LRS in improving the laying performance and intestinal morphology and microecological environment.

Determination of bacterial abundance and communities in the nipple drinking system of cascading cage layer houses.

Water quality is critical for egg production and animal health in commercial layer housing systems. To investigate microbial contamination in nipple drinking system in layer houses, the bacterial abundance and communities in water pipes and V-troughs on different tiers (e.g., 1st, 3rd, 5th, and 7th tiers) of a layer house with 8 overlapping cage tiers were determined using qRT-PCR and 16S rRNA sequencing. The water bacterial abundance (i.e., genome 16S rDNA copy number, WBCN) in water pipes and V-troughs did not significantly differ among tiers, but they were 46.77 to 1905.46 times higher in V-troughs than that in water pipes (P < 0.05) for each tier. Illumina sequencing obtained 1,746,303 effective reads from 24 water samples in V-troughs of 4 tiers (six samples from each tier). Taxonomic analysis indicated that the 1st and 5th tiers were predominated by Proteobacteria, Actinobacteria, Firmicutes and Bacteroidetes, while the 3rd and 7th tiers were predominated by Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria. The top four genera were Acinetobacter, Streptococcus, Rothia and Comamonas among measured tiers. The high bacterial abundance and bacterial OTUs of water in the V-troughs reflect poor water quality, which may adversely affect growth and health of laying hens. Therefore, it is suggested that water quality in the V-tough should be checked more frequently in commercial layer houses.

Effects of Different Non-Cage Housing Systems on the Production Performance, Serum Parameters and Intestinal Morphology of Laying Hens.

This study investigated the effects of plastic-net housing system (NRS) and floor-litter housing system (LRS) on the production performance, serum parameters and intestinal morphology of Shendan laying hens. A total of 1200 30-week-old hens were randomly allocated to the NRS and LRS groups, each of which included five replicates with 120 chickens in each replicate. The experiment was conducted from 32 to 40 weeks of age. Indoor airborne parameters were measured every 2 weeks, and indoor ground contamination was measured monthly. The laying rate and mortality of hens were recorded daily, and egg quality traits and serum parameters were measured every 2 weeks. At 40 weeks of age, four birds per replicate from each experimental group were selected for intestinal morphological observation. The results showed that the airborne bacteria number in the LRS was significantly higher than that in the NRS (p < 0.05) for most of the experimental period (except at 32 and 38 weeks of age), and the bacterial numbers on the surfaces of the floor and floor eggs in the LRS were approximately 10 times higher than those in the NRS (p < 0.05). Compared with the LRS, the NRS improved the laying rate (p < 0.05), reduced serum malondialdehyde (MDA) (p < 0.05) and corticosterone (CORT) concentrations and increased serum glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities, indicating favourable effects on antioxidative status. The NRS was significantly associated with an increased villus height (VH), villus height to crypt depth ratio (VCR) in the small intestine (p < 0.05) and increased VCR in the caecum (p < 0.05). Overall, the lower rate of bacterial contamination in the NRS than in the LRS indicated better environmental hygiene. The NRS enhanced the laying performance and antioxidant capacity of hens and was superior to the LRS in improving intestinal health. The current findings support the advantages of the NRS for the health and welfare of Shendan chickens during the peak laying period.

Effect of the Pellet and Mash Feed Forms on the Productive Performance, Egg Quality, Nutrient Metabolism, and Intestinal Morphology of Two Laying Hen Breeds.

One of the most important factors that determine feed utilization by chickens is the feed form. Although it is generally believed that pellet diets have a positive effect on chicken growth, there are some studies that have indicated no difference between pellet and mash on chickens performance. This study was conducted to assess the effects of feed form on production performance, egg quality, nutrient metabolism and intestinal morphology in two breed laying hens. Two hundred and sixteen 25-week-old Hy-Line brown (n = 108) and Hy-Line grey (n = 108) hens were selected. Each breed was randomly allocated into two treatments with 6 replications (9 birds in each replication), which were fed mash and pellet diets, respectively. Production performances were recorded daily and egg quality traits were measured every two weeks. At 42 weeks of age, one bird per replication from each experimental group was selected for metabolism determination and intestine morphology observation. Compared with mash diets, pellet diets improved laying rate (p < 0.05), ADFI (average daily feed intake, p < 0.05), egg weight, shell strength, yolk proportion and Haugh unit (p < 0.05) in both breeds and reduced the FCR (feed conversion ratio, p < 0.05) in Hy-Line grey. The apparent digestibility of DM% (dry matter) and CP% (crude protein) were significantly higher (p < 0.05) in both breed laying hens fed pellet than those fed mash. The apparent digestibility of P% (phosphorus) and Ca% (calcium) was higher in Hy-Line grey fed pellet and was higher in Hy-Line brown fed mash. Compared to mash diets, pellet diets increased the VH (villus height), CD (crypt depth) and VCR (ratio of villus height to crypt depth) of the small intestine of Hy-Line grey, and increased the VH and CD of duodenum and ileum of Hy-Line brown. Overall, pellet diets improved production performance and nutrition metabolism through positive changes in the laying rate, feed intake, egg albumen quality and apparent digestibility of laying hens. The current findings provided support for the advantages of feeding pellets during the peak egg laying period for the two popular laying hen strains, Hy-Line brown and Hy-Line grey.

Effect of a Large-sized Cage with a Low Metabolizable Energy and Low Crude Protein Diet on Growth Performance, Feed Cost, and Blood Parameters of Growing Layers.

Effects of a large-sized cage with a low metabolizable energy and low crude protein (LME-LCP) diet on growth performance, feed cost, blood biochemistry, and antibody response of growing layers were evaluated. A total of 668 one-day-old female Gushi chicks were randomly allocated into three different cages, namely, large, medium, and small cages, referred to as Group A, Group B, and Group C, respectively, and fed LME-LCP diets. A fourth group of birds raised in small cages and fed a standard diet (STD) was designated Group D i.e. the control. Equal stocking densities were maintained among the four groups throughout the experiment, from 50-20 birds/m2. Large cages with LME-LCP diet (Group A) increased the shank length and girth as chicks grew, improved the activities of serum creatine kinase, and reduced serum triglyceride and cholesterol concentrations. The total feed intake in birds from Group A was higher than those from the other groups at every stage. The total cost (rmb/bird) of feed was 6.70% lower in Group A than that in Group D, which indicated the cost-effectiveness of large cages. In conclusion, large cages with LME-LCP diets have positive effects on body weight, shank growth, and serum biochemical indices of growing Gushi chicks, and can reduce feed costs.

Effects of poor plumage conditions on egg production, antioxidant status and gene expression in laying hens.

The objective of this study was to investigate the effects of poor plumage conditions on production performance, antioxidant status and gene expression in laying hens. Two hundred ten 54-week-old laying hens with similar body weights were assigned into two groups based on plumage conditions (the poor plumage conditions (PPC) group and the control group). All the birds had free access to water and crumbled feed, and received the same management in step cages. Compared with hens in the control group, the hens in the PPC group consumed more feed and produced lighter eggs (P < 0.05). Hens in the PPC group showed lower serum concentrations of glutathione peroxidase and total antioxidant capacity and higher malondialdehyde content than those in the control group. The eggshell breaking strength was lower in the PPC group than in the control group (P < 0.05). The eggshell shape index and yolk colour in the PPC group were significantly higher than those in the control group. The mRNA expression level of HTR2C in the neck skin and that of IL-2 in the liver and breast muscle were higher in the PPC group than in the control group (P < 0.05). The results indicated that PPC may increase feed consumption and influence egg quality, antioxidant status and gene expression in laying hens.

A clear bias in parental origin of de novo pathogenic CNVs related to intellectual disability, developmental delay and multiple congenital anomalies.

Copy number variation (CNV) is of great significance in human evolution and disorders. Through tracing the parent-of-origin of de novo pathogenic CNVs, we are expected to investigate the relative contributions of germline genomic stability on reproductive health. In our study, short tandem repeat (STR) and single nucleotide polymorphism (SNP) were used to determine the parent-of-origin of 87 de novo pathogenic CNVs found in unrelated patients with intellectual disability (ID), developmental delay (DD) and multiple congenital anomalies (MCA). The results shown that there was a significant difference on the distribution of the parent-of-origin for different CNVs types (Chi-square test, p = 4.914 × 10-3). An apparently paternal bias existed in deletion CNVs and a maternal bias in duplication CNVs, indicating that the relative contribution of paternal germline variations is greater than that of maternal to the origin of deletions, and vice versa to the origin of duplications. By analyzing the sequences flanking the breakpoints, we also confirmed that non-allelic homologous recombination (NAHR) served as the major mechanism for the formation of recurrent CNVs whereas non-SDs-based mechanisms played a part in generating rare non-recurrent CNVs and might relate to the paternal germline bias in deletion CNVs.

WDR73 missense mutation causes infantile onset intellectual disability and cerebellar hypoplasia in a consanguineous family.

Galloway-Mowat syndrome (GMS) is a very rare autosomal-recessive disorder characterized by nephrotic syndrome associated with microcephaly, and various central nervous system abnormalities, mostly cerebral hypoplasia or cerebellar atrophy, intellectual disability and neural-migration defects. WDR73 is the only gene known to cause GMS, and has never been implicated in other disease. Here we present a Chinese consanguineous family with infantile onset intellectual disability and cerebellar hypoplasia but no microcephaly. Whole exome sequencing identified a WDR73 p.W371G missense mutation. The mutation is confirmed to be segregated in this family by Sanger sequencing according to a recessive inheritance pattern. It is predicted to be deleterious by multiple algorithms and affect highly conserved site. Structural modeling revealed conformational differences between the wild type protein and the p.W371G protein. Real-time PCR and Western blotting revealed altered mRNA and protein levels in mutated samples. Our study indicates the novel WDR73 p.W371G missense mutation causes infantile onset intellectual disability and cerebellar hypoplasia in recessive mode of inheritance. Our findings imply that microcephaly is a variable phenotype in WDR73-related disease, suggest WDR73 to be a candidate gene of severe intellectual disability and cerebellar hypoplasia, and expand the molecular spectrum of WDR73-related disease.

[Genetic diagnosis and analysis for two cases of ring chromosome 22].

OBJECTIVE: To confirm the genetic diagnosis of two patients with ring chromosome 22 syndrome and investigate the mechanism underlying the formation of r(22) and potential genetic causes for the clinical phenotypes. METHODS: Cytogenetic and molecular analyses using standard G-banding, fluorescence in situ hybridization and single nucleotide polymorphism array (SNP array) were performed. RESULTS: For case 1, the karyotype was 46,XY,r(22)(p11q13). SNP array has identified a 7.0 Mb heterozygous deletion at 22q13.2q13.33. For case 2, the karyotype was 46,XY,r(22)(p11q13)[84]/45,XY,-22[6]; SNP array has detected a heterozygous microdeletion of 1.6 Mb at 22q13.33. CONCLUSION: With combined application of genetic testing, 2 cases of r(22) syndrome were diagnosed, which has improved the understanding of the genotype-phenotype correlation of r(22).

Chromosomal analysis of blastocysts from balanced chromosomal rearrangement carriers.

Balanced chromosomal rearrangements (CRs) are among the most common genetic abnormalities in humans. In the present study, we have investigated the degree of consistency between the chromosomal composition of the blastocyst inner cell mass (ICM) and trophectoderm (TE) in carriers with balanced CR, which has not been previously addressed. As a secondary aim, we have also evaluated the validity of cleavage-stage preimplantation genetic diagnosis (PGD) based on fluorescence in situ hybridization (FISH) of blastocysts from CR carriers. Blastocyst ICM and TE were screened for chromosomal aneuploidy and imbalance of CR-associated chromosomes based on whole-genome copy number variation analysis by low-coverage next-generation sequencing (NGS) following single-cell whole-genome amplification by multiple annealing and looping-based amplification cycling. The NGS results were analyzed without knowledge of cleavage-stage FISH results. NGS results for blastocyst ICM and TE from CR carriers were 86.49% (32/37) consistent. Of the 1702 (37 × 46) chromosomes examined, 99.47% (1693/1702) showed consistency. However, only 40.0% (18/45) of all embryos had consistent results for chromosomes involved in CR, as determined by blastocyst NGS and cleavage-stage FISH. Of the 85 CR-affected chromosomes analyzed by FISH, 37.65% (32/85) were incongruous with NGS results, with 87.5% (28/32) showing imbalanced composition by FISH but balanced composition by NGS. These results indicate that chromosomal composition of blastocyst ICM and TE in balanced CR carriers is highly consistent, and that PGD based on cleavage-stage FISH is inaccurate; therefore, using blastocyst TE biopsies for NGS-based PGD is recommended for identifying chromosomal imbalance in embryos from balanced CR carriers.

Novel and reported APC germline mutations in Chinese patients with familial adenomatous polyposis.

OBJECTIVE: Familial adenomatous polyposis (FAP) is mainly caused by germline mutations in the adenomatous polyposis coli (APC) gene. This study aimed to detect pathogenic variants in five Chinese FAP families and review all previously reported pathogenic variants of APC gene in Chinese population. METHODS: Five non-consanguineous FAP families and 100 unrelated ethnicity-matched controls were included in the study. Sanger sequencing was performed to screen for APC coding and splicing variants. Chinese and English literature on APC germline mutations were reviewed to compile the mutation spectrum of APC gene in Chinese FAP patients. RESULTS: One pathogenic variant was detected in each family for the five pedigrees we tested. Three variants (c.3183_3187delACAAA, c.2626C>T and c.1312+1G>A) were previously reported as pathogenic. The other two variants were novel: c.794_795insG/p.Val266SerfsTer11 and c.2142_2143insG/p.His715AlafsTer19. They are absent from public databases (1000 Genomes, dbSNP, ESP and ExAC) and 100 normal controls, and are classified as pathogenic based on the new ACMG/AMP variant classification guidelines. Literature review and current study revealed a total of 82 different pathogenic variants from 127 Chinese FAP families. Among these families, 83 families had frameshift variants (65.35%), 26 with nonsense variants (20.47%), six with splice site variants (4.72%), three with missense variants (2.36%) and nine with large deletion or duplication variants (7.09%). Apart from the two previously reported mutation hotspots c.3927_3931delAAAGA (20.47%) and c.3183_3187delACAAA (7.09%), c.847C>T/p.Arg283Ter variant occurred with a frequency of 3.15% (4 out of 127) in Chinese FAP patients. CONCLUSIONS: We reported two novel pathogenic variants. The comprehensive compilation of variants and comparison revealed largely similar mutation spectrum between Chinese and Western patient populations. Some unique features noticed in Chinese patient population may help to better understand the pathogenesis of FAP.

Clinical and molecular investigation in Chinese patients with glutaric aciduria type I.

Glutaric aciduria type I (GA-I) is a rare autosomal recessive metabolic disorder caused by deficiency of glutaryl-CoA dehydrogenase (GCDH), leading to an abnormal metabolism of lysine, hydroxylysine and tryptophan. It results in accumulations of glutaric acid, 3-hydroxyglutaric acid and glutaconic acid. Clinical features include the sudden onset of encephalopathy, hypotonia and macrocephaly usually before age 18months. Here we report five cases of GA-I confirmed with mutation analysis. GCDH gene mutations were identified in all five probands with GA-I. Three of them had compound heterozygous mutations and two had homozygous mutations. Mutations of two alleles (c.334G>T and IVS11-11A>G) were novel and both of them were confirmed to be splice site mutations by reverse transcription PCR.