Whole exome sequencing approach for identification of the molecular etiology in pediatric patients with hematuria.

BACKGROUND: Hematuria is a common condition in clinical practice of pediatric patients. It is related to a wide spectrum of disorders and has high heterogeneity both clinically and genetically, which contributes to challenges of diagnosis and lead many pediatric patients with hematuria not to receive accurate diagnosis and early management. METHODS: In this single center study, 42 children with hematuria were included in Tianjin Children's Hospital between 2019 and 2020. We analyzed the clinical information and performed WES (Whole exome sequencing) for all cases. Then the classification of identified variants was performed according to the American College of Medical Genetics and Genomics (ACMG) guidelines for interpreting sequence variants. For the fragment deletion, qPCR was performed to validate and confirm the inherited pattern. RESULTS: For the 42 patients, 16 cases had gross hematuria and 26 had microscopic hematuria. Molecular genetic causes were uncovered in 9 (21.4%) children, including 7 with Alport syndrome (AS), one with polycystic nephropathy and one with lipoprotein glomerulopathy. The genetic causes for other patients were not related with hematuria. CONCLUSIONS: WES is a rapid and effective way to evaluate patients with hematuria. The analysis of genotype-phenotype correlations of patients with AS indicated that severe variants were associated with early kidney failure. Secondary findings were not rare in Chinese children, thus the clinician should pay more attention to the clinical interpretation of sequencing results and properly interaction with patients and their family.

[Predictive value of peripheral blood lymphocyte subsets for children with intravenous immunoglobulin-resistant Kawasaki disease]. / å¤–å‘¨è¡€æ·‹å·´ç»†èƒžäºšç¾¤å¯¹é™è„‰æ³¨å°„å ç–«çƒè›‹ç™½æ— ååº”åž‹å·å´Žç— æ‚£å„¿çš„é¢„æµ‹ä»·å€¼.

OBJECTIVES: Based on peripheral blood lymphocyte subsets and common laboratory test indexes, this study aimed to construct a predictive scoring system for intravenous immunoglobulin (IVIG)-resistant Kawasaki disease (KD). METHODS: Children hospitalized in Tianjin Children's Hospital from January 2021 to March 2023 were included in the study (185 cases of IVIG-sensitive KD and 41 cases of IVIG -resistant KD). Forty-six healthy children matched for age and gender were selected as controls. The relative percentage and absolute counts of peripheral lymphocyte subsets were measured by flow cytometry. Multivariate logistic regression was used to identify the predictive factors for IVIG-resistant KD and to construct a predictive scoring system for predicting IVIG-resistant KD. RESULTS: The multivariate logistic regression analysis showed that CD4+ T cell absolute count, natural killer cell absolute count, serum sodium level, globulin level, and total bilirubin level were identified as predictive factors for IVIG-resistant KD (P<0.05). The predictive scoring system based on these factors achieved a sensitivity of 70.7% and a specificity of 83.8% in predicting IVIG-resistant KD. CONCLUSIONS: Peripheral blood lymphocyte subsets can serve as predictive indicators for IVIG-resistant KD in children. The introduction of this indicator and the establishment of a scoring system based on it can provide a higher accuracy in predicting IVIG-resistant KD in children.

Comprehensive systematic review and meta-analysis of the association between common genetic variants and autism spectrum disorder.

BACKGROUND: Autism spectrum disorder (ASD) is neurodevelopmental disorder characterized by stereotyped behavior and deficits in communication and social interactions. To date, numerous studies have investigated the associations between genetic variants and ASD risk. However, the results of these published studies lack a clear consensus. In the present study, we performed a systematic review on the association between genetic variants and ASD risk. Meanwhile, we conducted a meta-analysis on available data to identify the association between the single nucleotide polymorphisms (SNPs) of candidate genes and ASD risk. METHODS: We systematically searched public databases including English and Chinese from their inception to August 1, 2022. Two independent reviewers extracted data and assessed study quality. Odds ratio and 95 % confidence interval were used as effect indexes to evaluate the association between the SNPs of candidate genes and the risk of ASD. Heterogeneity was explored through subgroup, sensitivity, and meta-regression analyses. Publication bias was assessed by using Egger's and Begg's tests for funnel plot asymmetry. In addition, TSA analysis were performed to confirm the study findings. RESULTS: We summarized 84 SNPs of 32 candidate genes from 81 articles included in the study. Subsequently, we analyzed 16 SNPs of eight genes by calculating pooled ORs, and identified eight significant SNPs of contactin associated protein 2 (CNTNAP2), methylentetrahydrofolate reductase (MTHFR), oxytocin receptor (OXTR), and vitamin D receptor (VDR). Results showed that seven SNPs, including the CNTNAP2 rs2710102 (homozygote, heterozygote, dominant and allelic models) and rs7794745 (heterozygote and dominant models), MTHFR C677T (homozygote, heterozygote, dominant, recessive and allelic models) and A1298C (dominant and allelic models), OXTR rs2254298 (homozygote and recessive models), VDR rs731236 (homozygote, dominant, recessive and allelic models) and rs2228570 (homozygote and recessive models), were showed to be correlated with an increased ASD risk. By contrast, the VDR rs7975232 was correlated with a decreased the risk of ASD under the homozygote and allelic models. CONCLUSION: Our study summarized research evidence on the genetic variants of ASD and provides a broad and detailed overview of ASD risk genes. The C677T and A1298C polymorphisms of MTHFR, rs2710102 and rs7794745 polymorphisms of CNTNAP2, rs2254298 polymorphism of OXTR, and rs731236 and rs2228570 polymorphisms of VDR were genetic risk factors. The rs7975232 polymorphism of VDR was a genetic protective factor for ASD. Our study provides novel clues to clinicians and healthcare decision-makers to predict ASD susceptibility.

Potential predictive value of CT radiomics features for treatment response in patients with COVID-19.

INTRODUCTION: This study aims to explore the predictive value of CT radiomics and clinical characteristics for treatment response in COVID-19 patients. METHODS: Data were collected from clinical/auxiliary examinations and follow-ups of COVID-19 patients. Whole lung radiomics feature extraction was performed at baseline chest CT. Radiomics, clinical, and combined features (nomogram) were evaluated for predicting treatment response. RESULTS: Among 36 COVID-19 patients, mild, common, severe, and critical disease symptoms were found in 1, 21, 13, and 1 of them, respectively. Twenty-five (1 mild, 18 common, and 6 severe) patients showed a good response to treatment and 11 poor/fair responses. The clinical classification (p = 0.025) and serum creatinine (p = 0.010) on admission and small area emphasis (p = 0.036) from radiomics analysis significantly differed between the two groups. Predictive models were constructed based on the radiomics, clinical features, and nomogram showing an area under the curve of 0.651, 0.836, and 0.869, respectively. The nomogram achieved good calibration. CONCLUSION: This new, non-invasive, and low-cost prediction model that combines the radiomics and clinical features is useful for identifying COVID-19 patients who may not respond well to treatment.

Eye movements of second language learners when reading spaced and unspaced Chinese texts.

Unlike English, Chinese does not have interword spacing in written texts, which poses difficulties for Chinese-as-a-second-language (CSL) learners' identification of word boundaries and affects their reading comprehension and vocabulary acquisition. The eye-movement literature has suggested that interword spacing is important in alphabetic languages; examining languages that lack interword spaces such as Chinese, thus, may help to inform theoretical accounts of eye-movement control and word identification during reading. Research investigating the interword spacing effect in reading Chinese showed that adding spacing facilitated CSL learners' reading comprehension and speed as well as vocabulary learning. However, the bulk of this research mainly looked at the learning outcomes (off-line measures), with few studies focusing on L2 learners' reading processes. Building on this background, this study seeks to provide a descriptive perspective of the eye movements of CSL learners. In this study, 24 CSL learners with intermediate Chinese proficiency were recruited as the experimental group, and 20 Chinese native speakers were recruited as the control group. The EyeLink 1,000 eye tracker was used to record their reading of four segmentation conditions of Chinese texts, namely, no space condition, word-spaced condition, non-word-spaced condition, and pinyin-spaced condition. Results show that: (1) CSL learners with intermediate Chinese proficiency generally spent less time reading Chinese texts with spaces between words, and they showed more gazes and regressions when reading texts without spaces; (2) Non-word-spaced texts and Pinyin-spaced texts interfere with CSL learners' reading process; and (3) Intermediate CSL learners show consistent eye movement patterns in the normal no-space condition and word-spaced condition. I conclude that word boundary information can effectively guide CSL learners' eye movement behaviors and eye saccade planning, thus improving reading efficiency.

Agency as power: An ecological exploration of an emerging language teacher leaders' emotional changes in an educational reform.

Teacher emotion, an important aspect of language teacher psychology (LTP), has recently drawn growing attention in language teacher development studies. Previous research has shown that language teachers, typically pressured by heavy workloads, may face emotional challenges from multiplied sources, especially in the context of educational changes such as curriculum reform and the COVID-19 emergency. Current literature on teachers' emotions largely centers around ordinary language teachers, with teacher leaders whose agentic actions often exert greater influence on the effectiveness of educational changes rarely examined. Situated in a top-tier research university that has been promoting an English for Academic Purposes reform to enhance its science students' multilingual competence in academic contexts, this longitudinal case study tracked the emotional trajectory of an English teacher, Lea, for 5 years. Adopting an ecological perspective, our study confirms that language teachers' emotions vary across the reform ecosystems and extends the current inquiry by conceptualizing the intricately interrelated teacher emotion, agency, power, and identity as dynamic constructs. This study also reveals how the reform-inflected emotional changes were associated with Lea's EAP teacher and teacher leader identity construction, with both identities reinforcing each other, which to some extent reconciled Lea's emotional tensions. Our study bears significant implications for language teachers involved in educational reform, teacher leaders, and school administrators.

Clinical and Magnetic Resonance Imaging Characteristics of Pediatric Acute Disseminating Encephalomyelitis With and Without Antibodies to Myelin Oligodendrocyte Glycoprotein.

Background: Myelin oligodendrocyte glycoprotein-immunoglobulin G (MOG-IgG)-associated disorders (MOGADs) have been considered as a new inflammatory disease entity of the central nervous system (CNS) and have heterogeneous clinical and imaging presentations. Acute disseminated encephalomyelitis (ADEM) is one of the most important phenotypes. Our research is aimed to compare the clinical and magnetic resonance imaging (MRI) characteristics of ADEM with or without MOG-IgG in pediatric-acquired demyelinating syndromes (ADSs). Methods and Results: We retrospectively reviewed the clinical characteristics, MRI features, and outcomes of pediatric patients with ADSs from March 2017 to February 2021 in our center. MOG-IgG was analyzed by transfected cell-based assay (CBA). Among 46 children with ADEM, 21 children (11 girls and 10 boys) were positive for MOG-IgG. Headache, fever, vomiting, vertigo, ataxia, and decreased muscle strength were common in all enrolled children. No significant difference existed in demographic characteristics, symptoms at an initial episode, or laboratory cerebrospinal fluid (CSF) findings between children with MOG-IgG and children without MOG-IgG. For children with MOG-IgG seropositive ADEM, cerebral MRI showed widespread, poorly demarcated bilateral lesions, especially in cortical and subcortical white matter, and spinal MRI often showed lesions spanning more than three segments. The significant difference in MRI features between the two groups was the presence of lesions in the thalamus and cortical area (p < 0.05). Most children in both groups showed clinical improvement 1 week after immunotherapy and achieved recovery during their hospital stay. Three children with MOG-IgG and four children without MOG-IgG had one or more relapsing courses with median interattack intervals of 4 (range: 1-7) months and 10 (range: 1-24) months, respectively. New clinical symptoms and lesions on cerebral and spinal MRI were found during relapsing courses in two groups. No recurrences were recorded 6-51 months after each patient's last episode. Conclusions: There was no significant difference in clinical characteristics between ADEM children with MOG-IgG and ADEM children without MOG-IgG. For children with MOG-IgG seropositive ADEM, cerebral MRI showed large, bilateral lesions and spinal MRI often showed lesions spanning more than three segments. Children achieved a favorable outcome regardless of MOG-IgG serostatus.

Identification of two aberrant transcripts by RNA sequencing for a novel variant c.3354 + 5 G > A of MED12 in a Chinese girl with non-syndromic intellectual disability.

BACKGROUND: Missense variants in MED12 are associated with MED12-related disorders. We aimed to clarify the molecular level changes and underlying pathogenic mechanism of a female patient in our study. METHODS: We reported a Chinese girl with clinical characteristics similar to MED12-related disorders. Trio whole exome sequencing (WES) was performed to identify related pathogenic variant(s) and RNA sequencing (RNA-seq) was subsequently applied to evaluate the effect of identified variant(s) on mRNA splicing. Moreover, X-chromosome inactivation (XCI) assay based on AR and RP2 was performed to reveal the XCI pattern of the female patient. RESULTS: The proband manifested mainly as mental retardation and language impairment. Trio WES revealed a novel heterozygous variant c.3354 + 5 G > A in intron 23 of MED12. RNA-seq identified two aberrant transcripts. XCI assay on AR revealed a homozygous result, while XCI based on RP2 showed random pattern in peripheral blood. CONCLUSION: In conclusion, we identified a novel variant c.3354 + 5 G > A by WES combined with RNA-seq, which extends the spectrum of MED12 variants and provide a basis for further genetic counseling. According to the result of two aberrant transcripts by RNA-seq, we speculate that our patient's milder clinical feature may be the consequence of multiple different transcripts.

Alteration of Allergen Fold of Bos d 5 into a Hypoallergenic Vaccine for Immunotherapy of Cow's Milk Allergy.

BACKGROUND: Cow's milk allergy (CMA) is the most common IgE-mediated food allergy and Bos d 5 is the major allergen in cow's milk proteins. More than 60% of the patients with CMA are sensitized to this protein. METHODS AND RESULTS: A recombinant protein, encoded by a synthetic gene and consisting of reassembled Bos d 5 fragments, was expressed in E. coli strain BL21 (DE3) cells and purified to homogeneity. The B5M lacked relevant IgE-reactivity and allergenic activity compared with Bos d 5 in dot-blot and basophil activation assays. T-cell proliferation experiments demonstrated that B5M preserved the main T cell epitopes of Bos d 5. Immunization of rabbits with B5M induced protective IgG antibodies that blocked the binding of patients' IgE antibodies to the wild-type allergen and inhibited the degranulation of basophils induced by Bos d 5. CONCLUSION: Thus, we developed a new strategy, which was based on rational molecular reassembly for allergen-specific immunotherapy (AIT) of CMA and food allergy.

Phenotypic and genotypic analysis of children with methylmalonic academia: A single-center study in China and a recent literature review.

BACKGROUND: Methylmalonic acidemia (MMA) is a rare inherited metabolic disease caused by methylmalonyl-CoA deficiency or cobalamin metabolism disorder. It is mainly inherited in autosomal recessive mode. According to whether combined with homocysteinemia and the causative genes, it can be divided into many different subtypes. Early diagnosis and early treatment can significantly improve the prognosis. METHODS: The children with MMA diagnosed in Tianjin Children's Hospital from 2012 to 2020 were collected. All the children underwent comprehensive physical and laboratory examinations. The metabolites in blood and urine were screened by mass spectrometry. Sanger sequencing, Next-generation sequencing and methylation detection were used for gene detection. RESULTS: The detection rate of MMA was 0.20% in children with high-risk of inherited metabolic diseases. The three most common clinical phenotypes of children with MMA were respiratory / metabolic acidosis, global developmental delay and anemia, which were found in 36.00%, 33.33% and 30.67% of children respectively. The most common mutations of MMACHC gene in children with cblC were c.609G > A, c.658_660delAAG and c.80A > G, with frequencies of 34.09%, 13.64% and 13.64%, respectively. CONCLUSIONS: This research expands the study of phenotype and genotype of MMA in Chinese population, and can provide reference for clinical diagnosis and treatment of MMA.

Multi-Parametric Magnetic Resonance Imaging-Based Radiomics Analysis of Cervical Cancer for Preoperative Prediction of Lymphovascular Space Invasion.

BACKGROUND: Detection of lymphovascular space invasion (LVSI) in early cervical cancer (CC) is challenging. To date, no standard clinical markers or screening tests have been used to detect LVSI preoperatively. Therefore, non-invasive risk stratification tools are highly desirable. OBJECTIVE: To train and validate a multi-parametric magnetic resonance imaging (mpMRI)-based radiomics model to detect LVSI in patients with CC and investigate its potential as a complementary tool to enhance the efficiency of risk assessment strategies. MATERIALS AND METHODS: The model was developed from the tumor volume of interest (VOI) of 125 patients with CC. A total of 1037 radiomics features obtained from conventional magnetic resonance imaging (MRI), including a small field-of-view (sFOV) high-resolution (HR)-T2-weighted MRI (T2WI), apparent diffusion coefficient (ADC), T2WI, fat-suppressed (FS)-T2WI, as well as axial and sagittal contrast-enhanced T1-weighted MRI (T1c). We conducted a radiomics-based characterization of each tumor region using pretreatment image data. Feature selection was performed using the least absolute shrinkage and selection operator method on the training set. The predictive performance was compared with single variates (clinical data and single-layer radiomics signatures) analyzed using a receiver operating characteristic (ROC) curve. Three-fold cross-validation performed 20 times was used to evaluate the accuracy of the trained classifiers and the stability of the selected features. The models were validated by using a validation set. RESULTS: Feature selection extracted the six most important features (3 from sFOV HR-T2WI, 1 T2WI, 1 FS-T2WI, and 1 T1c) for model construction. The mpMRI-combined radiomics model (area under the curve [AUC]: 0.940) reached a significantly higher performance (better than the clinical parameters [AUC: 0.730]), including any single-layer model using sFOV HR-T2WI (AUC: 0.840), T2WI (AUC: 0.770), FS-T2WI (AUC: 0.710), ADC maps (AUC: 0.650), sagittal, and axial T1c values (AUC: 0.710, 0.680) in the validation set. CONCLUSION: Biomarkers using multi-parametric radiomics features derived from preoperative MR images could predict LVSI in patients with CC.

Computed tomography and magnetic resonance imaging findings of nasal chondromesenchymal hamartoma in a young child: a case report.

Nasal chondromesenchymal hamartoma (NCMH) is a rare destructive benign neoplasm that predominantly develops in infants and young children. The lesion is usually located in the nasal cavity, often in the adjacent paranasal sinuses and orbital region and especially in the ethmoid sinus. Because the imaging characteristics of NCMH often mimic the features of malignant tumors, it is clinically important to study the radiographic appearance of this disease. Therefore, we herein present the computed tomography and magnetic resonance imaging findings of NCMH occurring in a 7-year-old girl. The mass was resected via an endoscopic surgical approach and definitively diagnosed as NCMH based on histologic and immunohistochemical analysis. However, signs of tumor recurrence manifested 45 months after surgery. NCMH can be locally aggressive with an expansive and destructive radiographic appearance, which highly implies a malignant neoplasm. Hence, an accurate diagnosis is essential to avoid potentially harmful therapies, and detailed computed tomography or magnetic resonance imaging should be performed prior to surgery. Selective arterial embolization is also an important part of preoperative management because the degree of enhancement may not be adequate to determine the blood supply of the tumor. Moreover, complete radical excision cannot guarantee that the lesion will not recur.

Quantitative determination of human serum testosterone via isotope dilution ultra­performance liquid chromatography tandem mass spectrometry.

Quantification of testosterone serves an important role in the differential diagnosis of androgen­related endocrine diseases. Mass spectrometry exhibits higher accuracy and lower variability than immunoassays, especially at low testosterone concentrations. The present study developed and validated an isotope dilution ultra­performance liquid chromatography tandem mass spectrometry method for determination of human serum testosterone. The serum was equilibrated with an isotopic internal standard and treated with acidic buffer to release hormones from their binding proteins. Testosterone was extracted via two serial liquid­liquid extractions. In the first stage, the lipid fractions from an acidic buffer solution were isolated using ethyl acetate and n­hexane. The organic phase was evaporated and reconstituted in a basic buffer solution. In the second stage, the polar impurities of n­hexane extraction were removed. Total testosterone in serum was quantified via ultra­performance liquid chromatography tandem mass spectrometry in multiple reaction monitoring mode with positive electrospray ionization. The coefficient of variation of the method for intra­ and inter­assay was 2.13% (1.40­2.77%) and 3.44% (3.06­3.66%), respectively. The recovery ranged from 94.32 to 108.60% for different samples. The limit of detection was 0.50 ng/dl and the linear range was from 1.00 to 1,000.00 ng/dl. In addition, the extraction efficiency in three different levels of quality control of the serum ranged from 85.02 to 93.29%. Moreover, structural analogues were investigated and were not indicated to affect the quantification of testosterone. The present method may enable quantification of testosterone in a clinical setting with high precision and accuracy.

A light-initiated chemiluminescent assay for rapid quantitation of allergen-specific IgG4 in clinical samples.

BACKGROUND: An increase in allergen-specific IgG4 (sIgG4), which serves as a blocking antibody, is associated with acquisition of immune tolerance after immunotherapy. In this study, we developed a rapid, sensitive, and homogeneous immunoassay based on the light-initiated chemiluminescent assay (LICA) technology for quantifying allergen sIgG4 in serum samples. METHODS: Allergen sIgG4 was measured in vitro by incubating the sample with biotinylated allergens and chemiluminescent beads coated with anti-human IgG4 antibody, followed by the addition of streptavidin-coated sensitizer beads. Multiple tests were performed to optimize the working conditions of the LICA and evaluate its performance. RESULTS: We established the optimal concentration of biotinylated allergens (250 ng/mL), the optimal dilution range (1:8 for Gal d 1, Gal d 2 sIgG4 and 1:4 for Gal d 3, Gal d 4 sIgG4), and the optimal incubation time (20 min for Gal d 1, Gal d 2 sIgG4 and 40 min for Gal d 3, Gal d 4 sIgG4). The lower limit of quantification (LLOQ) was 0.261 ng/mL. The coefficient variation (CV) of the LICA was <10%. The assay was unaffected by general interfering substances at physiological concentrations. It exhibited excellent accuracy to detect allergen-sIgG4 in human serum. Additionally, we demonstrated that the levels of Gal d 1, Gal d 2, and Gal d 3-sIgG4 were significantly higher in the egg allergy group (p < .05), but no differences were found between the groups for Gal d 4-sIgG4. CONCLUSIONS: The LICA demonstrated satisfactory performance and can be used for quantifying allergen sIgG4 in clinical practice.

Competitive light-initiated chemiluminescent assay: using 5-α-dihydrotestosterone-BSA as competitive antigen for quantitation of total testosterone in human sera.

This paper described a homogeneous method, light-initiated chemiluminescent assay (LICA), for quantitation of total testosterone in human sera. The assay was bead based and built on a competitive-binding reaction format, in which 5-α-dihydrotestosterone (5-α-DHT) competed with the testosterone in serum samples in binding with biotinylated anti-testosterone antibody. The more testosterone in the serum sample, the less 5-α-DHT that bonded with biotinylated anti-testosterone antibodies. 5-α-DHT was coupled with emission beads (doped with thioxene derivatives and Eu(III) as a chemiluminescence emitter) via bovine serum albumin as a linker. Once streptavidin-coated sensitizer beads (modified with phthalocyanine as a photosensitizer) were added, the streptavidin/biotin reaction between 5-α-DHT-bound anti-testosterone antibody and sensitizer beads could bring emission and sensitizer beads together, which allowed energy transfer from sensitizer bead to emission bead. As such, an exciting light (680 nm) impinging on the sensitizer beads led to light emission at 520-620 nm by emission beads. The strength of the emitted light was inversely proportional to the testosterone in serum sample. The detection range of this assay was from 13.3 to 1200 ng/dL. The coefficient variation for intra- and inter-assay was lower than 15%. The recovery of this method ranged from 95.5 to 105.9% for different samples. Moreover, the LICA assay was highly specific with low cross-reactivity and interference. The concentration of testosterone from 58 serum samples analyzed by the LICA method significantly correlated (y = 0.97x + 1.87, R2 = 0.970, p < 0.001) with those obtained with the SIEMENS Centaur Xp System. Graphical abstract ᅟ.

RNA-Seq Analysis of Rice Roots Reveals the Involvement of Post-Transcriptional Regulation in Response to Cadmium Stress.

Widely-spread cadmium (Cd) pollution in the soil threatens both crop production and human health. How plants deal with the excess Cd are largely unknown. To evaluate the molecular mechanism by which plants respond to Cd stress, rice seedlings were treated with two concentrations of Cd and subjected to deep RNA sequencing. Comprehensive RNA-Seq analysis of rice roots under two gradients of Cd treatment revealed 1169 Cd toxicity-responsive genes. These genes were involved in the reactive oxygen species scavenging system, stress response, cell wall formation, ion transport, and signal transduction. Nine out of 93 predicted long non-coding RNAs (lncRNAs) were detected as Cd-responsive lncRNAs due to their high correlation with the Cd stress response. In addition, we analyzed alternative splicing (AS) events under different Cd concentrations. Four hundred and seventy-six differential alternatively spliced genes with 542 aberrant splicing events were identified. GO analysis indicated that these genes were highly enriched in oxidation reduction and cellular response to chemical stimulus. Real-time qRT-PCR validation analysis strengthened the reliability of our RNA-Seq results. The results suggest that post-transcriptional AS regulation may also be involved in plant responses to high Cd stress.

[Observation on gametophyte development of Cibotium barometz].

OBJECTIVE: To observe the spores germinating process of Cibotium barometz, and understand the growth principle provided for experience for indoor culturing and further research. METHOD: The spores of C. barometz were cultured both in inorganic medium and in the soil from original habitat, and the whole process of spores germination and the development of gametophytic were observed under microscope. RESULT: The spores germinated about 1-2 weeks after being sowed, and the type of germination belonged to Vittaria-type. The prothallial plates formed in 25 days after being sowed, while hairs developed after the formation of the prothallial plate. The gametophyte formed about 40 days after being sowed. But the type of mature prothalli was cordate. The antheridia formed in 60 days after inoculation, while the archegonia developed in 10 days after the formation of antheridia. CONCLUSION: Soil based indoor culturing of C. barometz spores is practical and can be used for cultivation of C. barometz.