[Analysis of copy number variation in AZF region of Y chromosome in patients with spermatogenic failure].

OBJECTIVE: To explore the characteristics of copy number variation (CNV) within the Y chromosome azoospermia factor (AZF) region in patients with spermatogenesis disorders in the Shenzhen area. METHODS: A total of 123 patients with spermatogenesis disorders who had visited Shenzhen People's Hospital from January 2016 to October 2022 (including 73 patients with azoospermia and 50 patients with oligozoospermia) and 100 normal semen males were selected as the study subjects. The AZF region was detected with multiplex ligation-dependent probe amplification (MLPA), and the correlation between the CNV in the AZF region and spermatogenesis disorders was analyzed using the chi-square test or Fisher's exact test. RESULTS: 19 CNV were detected among 53 patients from the 223 samples, including 20 cases (27.40%, 20/73) from the azoospermia group, 19 cases (38%, 19/50) from the oligozoospermia group, and 14 cases (14%, 14/100) from the normal control group. In the azoospermia, oligozoospermia, and normal control groups, the detection rates for CNV related to the AZFa region (including AZFab and AZFabc) were 5.48% (4/73), 2.00% (1/50), and 0 (0/100), respectively. The detection rates for the AZFb region (including the AZFbc region) were 6.85% (5/73), 0 (0/50), and 0 (0/100), respectively. The detection rates for gr/gr deletions in the AZFc region were 2.74% (2/73), 6.00% (3/50), and 9.00% (9/100), respectively, and those for b2/b4 deletions in the AZFc region were 2.74% (2/73), 10.00% (5/50), and 0 (0/100), respectively. The detection rates for complex rearrangements in the AZFc region were 6.85% (5/73), 18.00% (9/50), and 3.00% (3/100), respectively. Statistical analysis showed no significant difference in the detection rate of gr/gr deletions between the three groups (Fisher's Exact Test value = 2.712, P = 0.249); the differences in the detection rate of b2/b4 deletions between the three groups were statistically significant (Fisher's Exact Test value = 9.489, P = 0.002); the differences in the detection rate of complex rearrangements in the AZFc region between the three groups were statistically significant (Fisher's Exact Test value = 9.493, P = 0.006). In this study, a rare AZFa region ARSLP1 gene deletion (involving SY86 deletion) was detected in a patient with oligozoospermia. CONCLUSION: CNV in the AZFa and AZFb regions have a severe impact on spermatogenesis, but partial deletion in the AZFa region (ARSLP1 gene deletion) has a minor impact on spermatogenesis. The b2/b4 deletion and complex rearrangement in the AZFc region may be risk factors for male infertility. The gr/gr deletion may not serve as a risk factor for male infertility in the Shenzhen area.

[Prenatal diagnosis and genetic analysis of a fetus with partial deletion of Yq and mosaicism of 45,X].

OBJECTIVE: To carry out prenatal diagnosis and genetic analysis for a fetus with disorders of sex development (DSDs). METHODS: A fetus with DSDs who was identified at the Shenzhen People's Hospital in September 2021 was selected as the study subject. Combined molecular genetic techniques including quantitative fluorescence PCR (QF-PCR), multiplex ligation-dependent probe amplification (MLPA), chromosomal microarray analysis (CMA), quantitative real-time PCR (qPCR), as well as cytogenetic techniques such as karyotyping analysis and fluorescence in situ hybridization (FISH) were applied. Ultrasonography was used to observe the phenotype of sex development. RESULTS: Molecular genetic testing suggested that the fetus had mosaicism of Yq11.222qter deletion and X monosomy. Combined with the result of cytogenetic testing, its karyotype was determined as mos 45,X[34]/46,X,del(Y)(q11.222)[61]/47,X,del(Y)(q11.222),del(Y)(q11.222)[5]. Ultrasound examination suggested hypospadia, which was confirmed after elective abortion. Combined the results of genetic testing and phenotypic analysis, the fetus was ultimately diagnosed with DSDs. CONCLUSION: This study has applied a variety of genetic techniques and ultrasonography to diagnose a fetus with DSDs with a complex karyotype.

Biogeographic distribution patterns of algal community in different urban lakes in China: Insights into the dynamics and co-existence.

Urban lake ecosystems are significant for social development, but currently we know little about the geographical distribution of algal community in urban lakes at a large-scale. In this study, we investigated the algal community structure in different areas of urban lakes in China and evaluated the influence of water quality parameters and geographical location on the algal community. The results showed that obvious differences in water quality and algal communities were observed among urban lakes in different geographical areas. Chlorophyta was the dominant phylum, followed by cyanobacteria in all areas. The network analysis indicated that algal community composition in urban lakes of the western and southern area showed more variations than the eastern and northern areas, respectively. Redundancy analysis and structural equation model revealed that nutrients and pH were dominant environmental factors that affected the algal community, and they showed higher influence than that of iron, manganese and COD Mn concentration. Importantly, algal community and density exhibited longitude and latitude relationship. In general, these results provided an ecological insight into large-scale geographical distributions of algal community in urban lakes, thereby having potential applications for management of the lakes.

Identification of microRNAs and their Endonucleolytic Cleavaged target mRNAs in colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) ranks the third among the most common malignancies globally. It is well known that microRNAs (miRNAs) play vital roles in destabilizing mRNAs and repressing their translations in this disease. However, the mechanism of miRNA-induced mRNA cleavage remains to be investigated. METHOD: In this study, high-throughput small RNA (sRNA) sequencing was utilized to identify and profile miRNAs from six pairs of colorectal cancer tissues (CTs) and adjacent tissues (CNs). Degradome sequencing (DS) was employed to detect the cleaved target genes. The Database for Annotation, Visualization and Integrated Discovery (DAVID) software was used for GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis. RESULTS: In total, 1278 known miRNAs (clustered into 337 families) and 131 novel miRNAs were characterized in the CT and CN libraries, respectively. Of those, 420 known and eight novel miRNAs were defined as differentially expressed miRNAs (DEmiRNAs) by comparing the expression levels observed in the CT and CN libraries. Furthermore, through DS, 9685 and 202 potential target transcripts were characterized as target genes for 268 known and 33 novel miRNAs, respectively. It was further predicted that a total of 264 targeted genes for the 85 DEmiRNAs are involved in proteoglycans in cancer and the AMP-activated protein kinase signaling pathway. After systemic analysis of prognosis-related miRNA targets in those cancer-related signal pathways, we found that two targets ezrin (EZR) and hematopoietic cell-specific Lyn substrate 1 (HCLS1) had the potential prognostic characteristics with CRC regarding over survival (OS) or recurrence. CONCLUSION: In total, we found that endonucleolytic miRNA-directed mRNA cleavage occurs in CRC. A number of potential genes targeted by CRC-related miRNAs were identified and some may have the potential as prognosis markers of CRC. The present findings may lead to an improved better appreciation of the novel interaction mode between miRNAs and target genes in CRC.

A near infrared fluorescent probe based on ICT for monitoring mitophagy in living cells.

Mitophagy, the process in which cells degrade dysfunctional organelles and recycle their nutrient substances by lysosomes, plays a vital role in cell metabolism and physiology. Herein, we present a highly targeting and near-infrared (NIR) mitochondrion fluorescent probe, which can monitor the process of autophagy. The response mechanism of the probe is based on intramolecular charge transfer (ICT) for the detection of autophagy and real-time imaging of living cells. We designed a primary amine as a pH sensitizing group, and due to the ICT process, the probe exhibits green fluorescence, and when it is protonated the ICT process is broken, and the NIR fluorescence will be restored. Simultaneously, the green fluorescence of the probe disappears. This probe exhibits excellent selectivity, high sensitivity and clean responsiveness, which indicate that it can be applied for high-targeting and high-sensitive imaging of the process of autophagy in living systems.

[Analysis for 6-methyladenine modification of DNA in chorionic tissue from aborted fetuses with monosomy 21].

OBJECTIVE: To study the correlation of genome-wide distribution of 6-methyladenine (6mA) of DNA in chorionic tissues from abortuses with monosomy 21. METHODS: Genomic DNA was extracted from chorionic samples from four abortuses with monosomy 21 and four without. After quality and purity test, partial DNA was subjected to chromatin immunoprecipitation with anti-6mA antibody, and then identified by sequencing. The sequencing data was analyzed by using bioinformatic software for the difference in 6mA between the two groups. RESULTS: Analysis of read peaks suggested that the control group have much more 6mA genes (n=4607) compared with the experiment group (n=1059). For chromosome 21, this difference is even more pronounced (8032 vs. 1769). Above results suggested that the level of 6mA modification in monosomy 21 is low. Gene ontology enrichment analysis and KEGG pathway enrichment analysis indicated that the absence of 6mA genes in monosomy 21 is closely related to the growth and development of embryo. CONCLUSION: The 6mA modification of human genes may play a similar role to 5-methylcytosine (5mC) modification during the growth and development of embryos.

Abundance of antibiotic resistance genes and their association with bacterial communities in activated sludge of wastewater treatment plants: Geographical distribution and network analysis.

Wastewater treatment plants (WWTPs) are deemed reservoirs of antibiotic resistance genes (ARGs). Bacterial phylogeny can shape the resistome in activated sludge. However, the co-occurrence and interaction of ARGs abundance and bacterial communities in different WWTPs located at continental scales are still not comprehensively understood. Here, we applied quantitative PCR and Miseq sequence approaches to unveil the changing profiles of ARGs (sul1, sul2, tetW, tetQ, tetX), intI1 gene, and bacterial communities in 18 geographically distributed WWTPs. The results showed that the average relative abundance of sul1and sul2 genes were 2.08 × 10-1 and 1.32 × 10-1 copies/16S rRNA copies, respectively. The abundance of tetW gene was positively correlated with the Shannon diversity index (H'), while both studied sul genes had significant positive relationship with the intI1gene. The highest average relative abundances of sul1, sul2, tetX, and intI1 genes were found in south region and oxidation ditch system. Network analysis found that 16 bacterial genera co-occurred with tetW gene. Co-occurrence patterns were revealed distinct community interactions between aerobic/anoxic/aerobic and oxidation ditch systems. The redundancy analysis model plot of the bacterial community composition clearly demonstrated that the sludge samples were significant differences among those from the different geographical areas, and the shifts in bacterial community composition were correlated with ARGs. Together, these findings from the present study will highlight the potential risks of ARGs and bacterial populations carrying these ARGs, and enable the development of suitable technique to control the dissemination of ARGs from WWTPs into aquatic environments.

Integrated analysis of mRNA, microRNA and protein in systemic lupus erythematosus-specific induced pluripotent stem cells from urine.

BACKGROUND: In clinical practice, it is difficult to monitor the repeating relapse in patients who have been suffering from systemic lupus erythematosus (SLE). The underlying etiology remains largely unknown. METHODS: Aiming to understand the pathogenesis of SLE, a detailed study was conducted. Renal tubular cells-derived iPSCs were successfully obtained from the urine of SLE patients and healthy controls. With the purpose to identify simultaneous expression profiling of microRNA, mRNA and protein, Illumina HiSeq™ 2000 System and iTRAQ-coupled 2D LC-MS/MS analysis were utilized in systemic lupus erythematosus-specific induced pluripotent stem cells (SLE-iPSCs) and normal control-iPSCs (NC-iPSCs). The integration of multiple profiling datasets was realized since it could facilitate the identification of non-seed miRNA targets, as well as differentially expressed mRNAs and proteins. RESULTS: For this study, profiling datasets of 1099 differentially expressed mRNAs, 223 differentially expressed microRNAs and 94 differentially expressed proteins were integrated. In order to investigate the influence of miRNA on the processes of regulating mRNAs and proteins' levels, potential targets of differentially expressed mRNAs and proteins were predicted using miRanda, TargetScan and Pictar. Multiple profiling datasets were integrated to facilitate the identification of miRNA targets, as well as differentially expressed mRNAs and proteins. Through gene ontology (GO) analysis of differentially expressed mRNAs and proteins, biological processes that drive proliferation were identified, such as mRNA processing and translation. Western blot and Q-PCR confirmed AK4 protein and mRNA up-regulation. The findings also showed that TAGLN's protein and mRNA level were down-regulated in SLE-iPSCs, both miR-371a-5p and let-7a-5p in SLE-iPSC were down-regulated and verified using Q-PCR. The up-regulation of AK4 involved in nucleotide biosynthesis suggested a general acceleration of anabolic metabolism induced by down-regulated miR-371a-5p, which might contribute to SLE. CONCLUSION: Based on high throughput analysis, integrated miRNA, mRNA, and protein expression data were generated. Differentially expressed dates were also adopted in conjunction with in-silico tools to identify potential candidates for SLE-iPSCs. Representative miRNA, mRNA and proteins were verified. It was also expected that the knowledge gained from this study can be applied to assess the usefulness of pathogenesis and novel biomarker candidates of SLE, which may develop a new way for SLE diagnosis.

Effects of hydrogen peroxide on Scenedesmus obliquus: Cell growth, antioxidant enzyme activity and intracellular protein fingerprinting.

Hydrogen peroxide (H2O2) is an environmental-friendly algicide and it is widely used to control algal blooms in aquatic ecosystems. However, the response of algal cell metabolic characteristics and intracellular protein profile under H2O2 stress is still not well understood. In the present study, the green alga Scenedesmus obliquus was exposed to different concentrations of H2O2 (0, 2, 6, 8 and 10 mg L-1) to evaluate the changes in algal morphological, physiological, and proteomic features to H2O2 exposure. The results showed that 8 mg L-1 of H2O2 could effectively inhibit the cell growth and photosynthetic activity of S. obliquus including chlorophyll-a content and chlorophyll fluorescence parameters. The increased activities of superoxide dismutase (SOD) and catalase (CAT) observed in this study indicate that cells exposure to H2O2 caused oxidative stress. The metabolic activity of S. obliquus was significantly decreased by H2O2 treatment. In terms of proteomic analysis, 251 differentially expressed proteins (DEPs) were successfully identified. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed significant protein enrichment in the metabolic pathways, photosynthesis, ascorbic acid, and alginate metabolism and phenylpropane biosynthesis of S. obliquus. The analysis of protein-protein interaction system shows that the pathways of photosynthesis and metabolic pathways of S. obliquus were essential to resist oxidative stress. Taking together, these results shed new lights on exploring the cell physiological metabolism and intracellular protein mechanisms of H2O2 inhibition on algal blooms.

Combined effects of seasonality and stagnation on tap water quality: Changes in chemical parameters, metabolic activity and co-existence in bacterial community.

In drinking water distribution pipeline systems, the tap water quality is regulated by several biotic and abiotic factors, which can threaten the health of consumers. Stagnation is inevitable in the water distribution pipeline however, the combined effects of seasonal changes and stagnation on tap water quality are not well understood. Here, we investigated the seasonal variations in the chemical and biological quality of water after overnight stagnation for a period of one year. The results showed that the tap water quality deteriorated after overnight stagnation, with up to a 2.7-fold increase in the total iron concentrations. The total bacterial cell concentrations increased by 59-231% after overnight stagnation. The total cell and cell-bound adenosine triphosphate (ATP) of the stagnant water samples peaked in summer. In addition, Biolog analysis showed that the metabolic activities of microbes were higher in spring. The bacterial community based on Illumina Miseq DNA sequence analysis found that Proteobacteria dominated the drinking water bacterial community. The bacterial community structure varied significantly among different seasons, where the diversity and richness of the community were much higher in spring. Structural equation modeling (SEM) was constructed to determine the correlations between bacterial metabolic functions and the community structure. The redundancy analysis (RDA) indicated that the residual chlorine played a critical role in the construction of the bacterial community. Altogether, the overall findings from the present work provide novel insights into how the quality of tap water quality impacted by the seasonal changes and overnight stagnation.

A report on seven fetal cases associated with 15q11-q13 microdeletion and microduplication.

BACKGROUND: The 15q11-q13 region contains three breakpoints (BP1 to BP3), and copy number variations often occur in the region. AIMS: 15q11-q13 microdeletion and microduplication are usually associated with Prader-Willi and Angelman syndromes, respectively. It is not yet clear to what extent microdeletion and microduplication affect the physical health of the fetus and the child. In this study, we examined seven fetuses ranging in gestational age from 15 to 27 weeks. MATERIALS & METHODS: Detailed prenatal screening and laboratory examinations were performed, while karyotype analysis and chromosomal microarray analysis (CMA) of the amniotic fluid and umbilical cord blood were applied for genetic analysis. RESULTS: CMA analysis showed that four fetuses harbored a microdeletion and one fetus showed a microduplication at 15q11.2 BP1-BP2, two fetuses had a microdeletion at 15q11-q13 BP2-BP3, and one fetus had an additional microdeletion at 16p13.11. DISCUSSION: There is no clear standard for the clinical diagnosis of 15q11-q13 microdeletion and microduplication, some of them have clinical phenotypes or are clinically affected. CONCLUSION: Therefore, parents of such fetuses should be informed of the possibility of microdeletions or microduplications to mitigate the psychological burden, and medical consultation and assistance should be provided when communicating the results of the mid-gestation screening.

Charge-Mediated Co-assembly of Amphiphilic Peptide and Antibiotics Into Supramolecular Hydrogel With Antibacterial Activity.

Bacteria are the most common pathogens to cause infection of surgical sites, which usually induce severe postoperative morbidity and more healthcare costs. Inhibition of bacteria adhesion and colonization is an effective strategy to prevent the spread of infection at the surgical sites. Hydrogels have been widely used as promising antibacterial materials, due to their unique porous structure that could accommodate various antibacterial agents (e.g., antibiotics and cationic polymers with inherent antibacterial activity). Herein, inspired by the natural protein self-assembly, an amphiphilic peptide comprised of a hydrophobic naphthyl (Nap) acetyl tail and a hydrophilic peptide backbone was employed to construct supramolecular hydrogel for sustained release of the antibiotic polymyxin B. At neutral pH, the negatively charged amphiphilic peptide could form electrostatic attraction interaction with the positively charged polymyxin B, which could thus drive the ionized peptide molecules to get close to each other and subsequently trigger the self-assembly of the amphiphilic peptide into supramolecular hydrogel via intermolecular hydrogen bonding interaction among the peptide backbones and π-stacking of the hydrophobic Nap tails. More importantly, the electrostatic attraction interaction between polymyxin B and the amphiphilic peptide could ensure the sustained release of polymyxin B from the supramolecular hydrogel, leading to an effective inhibition of Gram-negative bacteria Escherichia coli growth. Combining the good biocompatibility of the amphiphilic peptide, the supramolecular hydrogel developed in this work shows a great potential for the surgical site infection application.

Geographical Patterns of Algal Communities Associated with Different Urban Lakes in China.

Urban lakes play an important role in drainage and water storage, regulating urban microclimate conditions, supplying groundwater, and meeting citizens' recreational needs. However, geographical patterns of algal communities associated with urban lakes from a large scale are still unclear. In the present work, the geographical variation of algal communities and water quality parameters in different urban lakes in China were determined. The water quality parameters were examined in the samples collected from north, central, south, and coastal economic zones in China. The results suggested that significant differences in water quality were observed among different geographical distribution of urban lakes. The highest total phosphorus (TP)(0.21 mg/L) and total nitrogen (TN) (3.84 mg/L) concentrations were found in XinHaiHu (XHH) lake, it also showed highest the nitrate nitrogen (NO3--N) (0.39 mg/L),total organic carbon(TOC) (9.77 mg/L), and COD Mn (9.01 mg/L) concentrations among all samples. Environmental and geographic factors also cause large differences in algal cell concentration in different urban lakes, which ranged from 4,700×104 to 247,800 ×104cell/L. Through light microscopy, 6 phyla were identified, which includes Chlorophyta, Bacillariophyta, Cyanophyta, Dinophyta, Euglenophyta, and Cryptophyta. Meanwhile, the heat map with the total 63 algal community composition at the genus level profile different urban lakes community structures are clearly distinguishable. Further analyses showed that the dominant genera were Limnothrixsp., Synedra sp., Cyclotella sp., Nephrocytium sp., Melosirasp., and Scenedesmussp. among all samples. The integrated network analysis indicated that the highly connected taxa (hub) were Fragilariasp.,Scenedesmus sp., and Stephanodiscus sp. The water quality parameters of NO3--N and NH4+-N had significant impacts on the structural composition of the algal community. Additionally, RDA further revealed distinct algal communities in the different urban lakes, and were influenced by NO2--N, Fe, and algal cell concentrations. In summary, these results demonstrate that the pattern of algal communities are highly correlated with geographic location and water quality on a large scale, and these results also give us further understanding of the complex algal communities and effectively managing eutrophication of urban lakes.

Differential expression profile study and gene function analysis of maternal foetal-derived circRNA for screening for Down's syndrome.

Recent studies have shown that circular RNAs (circRNAs) exhibit differential expression in certain diseases. However, to the best of our knowledge, maternal fetal-derived circRNAs and mRNAs associated with Down's syndrome (DS) have not yet been investigated. A total of 12 umbilical cord blood samples were collected from pregnant women, including six women carrying fetuses with DS (diagnosed by G-banding karyotype analysis), and six women carrying fetuses without DS. In addition, 12 peripheral blood samples were obtained from children, including six children with DS and six children without DS. Gene chip technology was used to screen for differentially expressed circRNAs and mRNAs in the cord blood samples, and were subsequently verified by reverse transcription-quantitative polymerase chain reaction in peripheral blood from the children to identify potential biomarkers. Furthermore, circRNA/microRNA (miRNA) interactions were predicted using Arraystar miRNA target prediction software. There was a significant difference in the expression of hsa_circRNA_103127, hsa_circRNA_103112 and hsa_circRNA_104907 between cord blood obtained from the women carrying fetuses with and without DS, and between peripheral blood obtained from children with and without DS (P<0.01). As hsa_circRNA_103112 exhibited significant differences in expression between cord blood obtained from the women carrying fetuses with and without DS and between peripheral blood obtained from children with and without DS, its corresponding gene, ubiquitin specific peptidase 25, may be involved in the pathogenesis of the condition. These results suggested that hsa_circRNA_103112 may be upregulated in individuals with DS, resulting in an expression imbalance of diploid genes through interactions among circRNA, miRNA and mRNA. Therefore, the level of hsa_circRNA_103112 in the peripheral blood of a pregnant woman may serve as potential biomarker of fetal DS during non-invasive prenatal screening.

Exploring a Moderate Fallow Scale of Cultivated Land in China from the Perspective of Food Security.

Food security remains a primary concern because of the large population and scarce land resources in China, and it is a core task to determine the appropriate proportion and scale of fallowing for fallow systems. The aim of this study was to systematically estimate the grain production potential (GPP) of existing and unexcavated cultivated land due to land use change from 1990 to 2017 and calculate the theoretical fallowing scale by using a population carrying capacity model. The reserved GPP from cultivated land to be excavated was 7470 × 104 t in China, and the GPP stored by the change in grain yield per unit, multiple crop index (MCI) decline, and agricultural structure adjustment were 921 × 104 t, 4321 × 104 t, and 7760 × 104 t, respectively, and the lost GPP caused by construction land expansion was 5287 × 104 t. The population carrying capacity of cultivated land in China was estimated to be 1.469 to 1.515 billion in 2017 on the basis of the national average living standard. The proportion of the population that could be fed more was between 6.28% and 9.54% depending on the number of people included, which provided an opportunity to implement the fallowing system in China. Meanwhile the proportions of the theoretical fallow scale were 6.28% and 9.54%, and the fallow scale ranged from 850 × 104 hm2 to 1296 × 104 hm2 under the premise of fully tapping the potential of cultivated land. In addition, taking the decline in MCI as an example, the grain yield reduction was equivalent to the grain yield of 829 × 104 hm2 of newly reclaimed cultivated land over the past 30 years, which saved 621.48 billion yuan. The costs and benefits when formulating relevant policies of land utilization should be considered, and exploiting the productive capacity of cultivated land that exists is better than reclaiming new cultivated land.

Targeted next generation sequencing identified novel loss-of-function mutations in MERTK gene in Chinese patients with retinitis pigmentosa.

BACKGROUND: Retinitis pigmentosa (RP) is one of the major types of hereditary retinal dystrophies with extreme genotypic heterogeneity. To date, more than 80 genes have been identified to be associated with RP in human. METHOD: Here, we presented a clinical genetic study of three Chinese man manifested with night vision blindness and complete loss of midperipheral visual field. All of these three probands have been identified with loss of both central vision and far peripheral visual field. Gradual loss of rod cells followed by subsequent loss of cone cells have been identified in these probands. Targeted next generation sequencing and Sanger sequencing have been performed to understand the pathogenic variants underlying the disease phenotype in these three unrelated Chinese probands. RESULTS: Targeted next generation sequencing and Sanger sequencing identified three homozygous novel mutations (c.1880C>A; c.1459_1460delGA, and c.392G>A) in the MERTK gene in these three unrelated Chinese proband. In the first proband, the identified mutation (c.1880C>A) leads to the formation of a premature stop codon followed by the formation of a truncated mer-tyrosine kinase (MERTK) protein (p.Ser627*) product which predicted to be disease causing. In the second proband, the identified deletion (c.1459_1460delGA) leads to the formation of a frameshift which also finally results in the formation of a truncated MERTK protein (p.Asp487Leufs*57) product which also predicted to be disease causing. In the third proband, the identified mutation (c.392G>A) leads to the formation of a premature stop codon followed by the formation of a truncated MERTK protein (p.Trp131*) product which also predicted to be disease causing. Hence, these three mutations are loss-of-function mutations. These three mutations were absent in unaffected family members and in 100 normal healthy controls. CONCLUSION: Our present study also demonstrates the significance of targeted next generation sequencing in determining the genetic basis of RP.

Chronic Trigeminal Nerve Stimulation Protects Against Seizures, Cognitive Impairments, Hippocampal Apoptosis, and Inflammatory Responses in Epileptic Rats.

Trigeminal nerve stimulation (TNS) has recently been demonstrated effective in the treatment of epilepsy and mood disorders. Here, we aim to determine the effects of TNS on epileptogenesis, cognitive function, and the associated hippocampal apoptosis and inflammatory responses. Rats were injected with pilocarpine to produce status epilepticus (SE) and the following chronic epilepsy. After SE induction, TNS treatment was conducted for 4 consecutive weeks. A pilocarpine re-injection was then used to induce a seizure in the epileptic rats. The hippocampal neuronal apoptosis induced by seizure was assessed by TUNEL staining and inflammatory responses by immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). The spontaneous recurrent seizure (SRS) number was counted through video monitoring, and the cognitive function assessed through Morris Water Maze (MWM) test. TNS treatment attenuated the SRS attacks and improved the cognitive impairment in epileptic rats. A pilocarpine re-injection resulted in less hippocampal neuronal apoptosis and reduced level of interleukin-1 beta (IL-1ß), tumor necrosis factor-α (TNF-α), and microglial activation in epileptic rats with TNS treatment in comparison to the epileptic rats without TNS treatment. It is concluded that TNS treatment shortly after SE not only protected against the chronic spontaneous seizures but also improved cognitive impairments. These antiepileptic properties of TNS may be related to its attenuating effects on hippocampal apoptosis and pro-inflammatory responses.

Quantitative proteomic analysis of Down syndrome in the umbilical cord blood using iTRAQ.

Down sydrome (DS) is a relatively frequent chromosomal disorder, which has no safe and effective method of prenatal diagnosis to date. The present study was designed to identify DS biomarkers. We quantified the changes in the umbilical cord blood protein levels between DS-affected and healthy (control) pregnant females using isobaric tags for relative and absolute quantification (iTRAQ) and Gene Ontology (GO) analysis. A total of 505 proteins were identified, and of these, five proteins showed significantly different concentrations between the DS and the control group. These proteins may thus be relevant to DS and constitute potential DS biomarkers.

Comparative proteomic analysis of membranous nephropathy biopsy tissues using quantitative proteomics.

Membranous nephropathy (MN) is a common cause of nephrotic syndrome in adults and the second leading cause of end-stage renal disease due to primary glomerulonephritis. The aim of the present study was to identify potential biomarkers of MN and further characterize these proteins by Gene Ontology (GO) analysis. Isobaric tags for relative and absolute quantification were used to compare the protein levels in tissues from MN patients and healthy individuals, and the combined samples were subsequently separated by specialized communications exchange. Mass spectrometry data acquisition was conducted using a 4800 Plus MALDI TOF/TOF tandem mass spectrometry device, and the results were subjected to statistical analysis. A total of 1,903 proteins were identified, with 423 proteins exhibiting a difference of >1.5-fold compared with the control group. Of these, 202 proteins were upregulated, while 221 proteins were downregulated. In conclusion, GO enrichment analysis revealed that the differentially expressed proteins were primarily mapped to the following GO terms: 'Immune response', 'immune effector process', 'activation of immune response' and 'positive regulation of immune system process'. The affected proteins may be associated with the pathogenesis of MN; thus, may represent candidate MN biomarkers.

Neuronal and astroglial TGFß-Smad3 signaling pathways differentially regulate dendrite growth and synaptogenesis.

To address the role of the transforming growth factor beta (TGFß)-Smad3 signaling pathway in dendrite growth and associated synaptogenesis, we used small inhibitory RNA to knockdown the Smad3 gene in either cultured neurons and or primary astrocytes. We found that TGFß1 treatment of primary neurons increased dendrite extensions and the number of synapsin-1-positive synapses. When Smad3 was knockdown in primary neurons, dendrite growth was inhibited and the number of synapsin-1-positive synapses reduced even with TGFß1 treatment. When astrocyte-conditioned medium (ACM), collected from TGFß1-treated astrocytes (TGFß1-stimulated ACM), was added to cultured neurons, dendritic growth was inhibited and the number of synapsin-1-positive puncta reduced. When TGFß1-stimulated ACM was collected from astrocytes with Smad3 knocked down, this conditioned media promoted the growth of dendrites and the number of synapsin-1-positive puncta in cultured neurons. We further found that TGFß1 signaling through Smad3 increased the expression of chondroitin sulfate proteoglycans, neurocan, and phosphacan in ACM. Application of chondroitinase ABC to the TGFß1-stimulated ACM reversed its inhibitory effects on the dendrite growth and the number of synapsin-1-positive puncta. On the other hand, we found that TGFß1 treatment caused a facilitation of Smad3 phosphorylation and translocation to the nucleus induced by status epilepticus (SE) in wild-type (Smad3(+/+)) mice, and this treatment also caused a promotion of γ-aminobutyric acid-ergic synaptogenesis impaired by SE in Smad3(+/+) as well as in Smad3(-/-) mice, but more dramatic promotion in Smad3(+/+) mice. Thus, we provide evidence for the first time that TGFß-Smad3 signaling pathways within neuron and astrocyte differentially regulate dendrite growth and synaptogenesis, and this pathway may be involved in the pathogenesis of some central nervous system diseases, such as epilepsy.