Expanding the Phenotypic and Genotypic Spectrum of ARFGEF1-Related Neurodevelopmental Disorder.

Mono-allelic loss-of-function variants in ARFGEF1 have recently caused a developmental delay, intellectual disability, and epilepsy, with varying clinical expressivity. However, given the clinical heterogeneity and low-penetrance mutations of ARFGEF1-related neurodevelopmental disorder, the robustness of the gene-disease association requires additional evidence. In this study, five novel heterozygous ARFGEF1 variants were identified in five unrelated pediatric patients with neurodevelopmental disorders, including one missense change (c.3539T>G), two canonical splice site variants (c.917-1G>T, c.2850+2T>A), and two frameshift (c.2923_c.2924delCT, c.4951delG) mutations resulting in truncation of ARFGEF1. The pathogenic/likely pathogenic variants presented here will be highly beneficial to patients undergoing genetic testing in the future by providing an expanded reference list of disease-causing variants.

Icariside II induces ferroptosis in renal cell carcinoma cells by regulating the miR-324-3p/GPX4 axis.

Icariside II (ICS II) is an active flavonoid having anti-tumor properties. However, the role of ICS II in renal cell carcinoma (RCC) and its underlying mechanisms have not been investigated to date. In this study, we demonstrated that ICS II inhibited proliferation, migration, and invasion of RCC cells. Furthermore, ferroptosis, a novel form of cell death, induced in RCC cells by ICS II, accompanied by accumulation of Fe2+, MDA (lipid peroxidation), and ROS (reactive oxygen species), and reduced GSH levels. The underlying mechanism was found to be the downregulation of GPX4, independent of p53, that occurs during ICS II-induced ferroptosis. Overexpression of GPX4 reversed the ferroptosis induced by ICS II. Moreover, ICS II treatment resulted in the upregulation of miR-324-3p, which directly targets GPX4. Overall, our results suggested that ICS II-induced ferroptosis via the miR-324-3p/GPX4 axis in RCC cells could be a promising therapeutic agent for RCC.

Effects of Levetiracetam on the Serum C-Reactive Protein in Children With Epilepsy: A Meta-Analysis.

This meta-analysis aims to evaluate the effect of levetiracetam on serum C-reactive protein (CRP) in children with epilepsy. Articles published up to April 15, 2021 were searched from Google Scholar databases, PubMed, Science Direct, Springer, Wiely, NIH and Baidu Scholar databases to analyzed the difference of serum CRP in epilepsy children compared to healthy controls, and the effect of levetiracetam on serum CRP in children with epilepsy was also assessed. All the included studies met the inclusion criteria. 103 publications were selected and eight articles were included in this study with sample size n = 246. The serum CRP level in childhood epilepsy was significantly higher than the healthy controls (pooled standardized mean difference (SMD): 6.930, 95% CI: 2.716-11.143, z = 3.22, p < 0.01). A significant level of between-study heterogeneity was found (τ2 = 17.911, Chi2 = 148.67, df = 3, p < 0.01, I2 = 98.0%). Besides, serum CRP level was significantly decreased by the treatment of levetiracetam in childhood epilepsy (pooled SMD: 3.505, 95% CI: 1.638-5.373, z = 3.68, p < 0.01). A significant level of between-study heterogeneity was found (τ2 = 4.346, Chi2 = 97.17, df = 4, p < 0.01, I2 = 95.9%). The funnel plot showed there was no significant publication bias in the meta-analysis. Serum CRP levels are upregulated in childhood epilepsy and reduced by levetiracetam in children with epilepsy.

Microcornea, iris and choroidal coloboma, and global developmental delay caused by TENM3 pathogenic variants in a Chinese patient.

BACKGROUND: Biallelic TENM3 pathogenic variants cause isolated or syndromic microphthalmia. Syndromic microphthalmia 15 (MCOPS15) is characterized by microphthalmia, coloboma, and developmental delay. Currently, only four cases of MCOPS15 have been reported and the clinical features varied among the patients indicating potential broad phenotypic spectrum. METHODS: The present case was a 6-month-old male at diagnosis. The patient exhibited long philtrum, large ears, bilateral ptosis, and nystagmus. Ophthalmic tests showed that he had microcornea, iris and choroidal coloboma. The patient presented with global developmental delay (GDD). Trio-whole exome sequencing and genome copy number sequencing were conducted to explore the disease-causing mutations. RESULTS: Exome sequencing and genome copy number sequencing showed the presence of L1471F and E661G compound mutations in TENM3, which were inherited from the mother and father, respectively. Sanger sequencing was conducted to verify association of the mutations with the disease in the present family. CONCLUSION: Two TENM3 variants were identified in a patient with Syndromic microphthalmia 15 in the present study. However, further studies should be conducted to explore the pathogenicity of the variants.

Effect of biofertilizer and wheat straw biochar application on nitrous oxide emission and ammonia volatilization from paddy soil.

Biofertilizer can improve soil quality, especially the microbiome composition, which potentially affect soil nitrogen (N) cycling. However, little is known about the responses of nitrous oxide (N2O) emission and ammonia (NH3) volatilization from biochar-amended paddy soil to the biofertilizer application. Therefore, we conducted a soil column experiment using four 240 kg N ha-1 (equivalent to 1.7 g N pot-1) treatments consisting of biofertilizer (3 t ha-1, equivalent to 21.2 g pot-1), biochar (7.5 t ha-1, equivalent to 63.6 g pot-1), and a mixture of biofertilizer and biochar at the same rate and a control (CK). The results showed that the N2O emissions and NH3 volatilizations were equivalent to 0.15-0.28% and 18.0-31.5% of rice seasonal N applied to the four treatments, respectively. Two treatments with biofertilizer and biochar individual amendment significantly increased (P < 0.05) the N2O emissions to same degree by 30.2%, while co-application of biochar and biofertilizer further increased the N2O emission by 74.4% compared to the control. The higher N2O emission was likely attributed to the increased gene copies of AOA, nirK, and nirS. Applying biofertilizer significantly increased (P < 0.05) NH3 volatilization by 24.7% relative to the control, while applying biochar had no influence on NH3 volatilization. Co-application of biofertilizer and biochar significantly decreased (P < 0.05) NH3 volatilization by 12.3% compared to the control. Overall, the net global warming potential based on NH3 and N2O in current study increased by 13.0-26.0% in both the individual- and co-application of biofertilizer and biochar. Interestingly, both individual- and co-applications of biofertilizer and biochar increased the rice grain yield by 16.5-38.3%. Therefore, applications of biofertilizer and biochar did not increase the GHGI. Particularly, the co-applying of them significantly lowered (P < 0.05) the GHGI by 15.2%. In conclusion, biofertilizer and biochar should be co-applied to achieve the goals of environment protection and food security.

[Clinical and genetic analysis of two patients with congenital neutropenia caused by ELANE gene mutation].

OBJECTIVE: To explore the clinical characteristics of congenital neutropenia caused by ELANE gene mutations. METHODS: Clinical manifestations, absolute blood neutrophil count, high-throughput exome sequencing for mutation screening, suspected locus Sanger sequencing verification, processes of diagnosis and treatment of two patients with congenital neutropenia caused by ELANE gene mutation were retrospectively analyzed. RESULTS: High-throughput sequencing has found that proband 1 has carried a heterozygous c.170C>T (p.Ala57Val) missense mutation in exon 2 of the ELANE gene, which was known to be pathological, and a heterozygous c.251T>G (p.Leu84Arg) mutation in exon 3 of proband 2, which was unreported previously. Sanger sequencing confirmed that neither mutation was inherited from their parents. CONCLUSION: ELANE mutation is an important cause for congenital neutropenia. Detection of new pathogenic variants has enriched the mutation spectrum of the ELANE gene.

[Genetic analysis of a male infant with Menkes disease].

OBJECTIVE: To carry out genetic testing for a male infant suspected for Menkes disease. METHODS: Genomic DNA of the proband and his parents were extracted and subjected to family trio whole exome sequencing (WES). Microduplication and microdeletion of the ATP7A gene were detected by multiplex ligation-dependent probe amplification (MLPA). Suspected variants were subjected to bioinformatic analysis and verified by Sanger sequencing. RESULTS: The proband was found to harbor a de novo c.1870 -13T>G variation of the ATP7A gene, which may alter a splice site and affect its protein product. CONCLUSION: The patient was diagnosed with Menkes disease due to the c.1870 -13T>G variant of the ATP7A gene. Whole exome sequencing of family trios is a powerful tool for the diagnosis of diseases with strong phenotypic heterogeneity.

Heavy metal accumulation, risk assessment and integrated biomarker responses of local vegetables: A case study along the Le'an river.

In this research, Ganzhou Chinese Cabbage (Brassica rapa pekinensis), Native Purple Garlic (Allium sativum L) and Leping Radish (Raphanus sativus L) widely planted and distributed along the Le'an River were chosen in the present study. Soil physical-chemical properties, nutrients contents as well as heavy metals elements accumulated in both soils and vegetables collected from 24 sites were analyzed by lab analysis combined with statistical method which was also used for calculation of contamination factor, pollution indexes and hazardous index. Heavy metals accumulation in soils were revealed with higher level, and copper and cadmium exceeded the background values by 8.82 and 16.73 times on average, which were also significantly related with the distribution of nonferrous metal processing enterprises. Heavy metal elements accumulated in vegetables were fully consistent with the finding of pollution characteristics in soils. Peroxidase biomarkers in vegetables, including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), reduced glutathione (GSH) and lipoperoxidation (as TBARS), as well as integrated biomarker responses (IBR) were determined to give a reliable response after exposing of contaminants. Heavy metals accumulation ability and biomarker responses for three vegetables were usually determined in the following decrease trend: Ganzhou Chinese Cabbage > Native Purple Garlic > Leping Radish. Compared with peroxidase biomarkers activities or contents of control site, all the measured biomarkers in polluted sites showed significantly responses, indicating potential relationship between pollutants stresses and biomarker responses. This study also revealed that the IBR values were coordinated well with the pollutants concentrations.

[Simultaneous inhibition of both hTERT and androgen receptor gene expression in LNCaP cells using single shRNA vector].

OBJECTIVE: To explore the feasibility of inhibition of hTERT and androgen receptor (AR) gene expression simultaneously in LNCaP cells by single shRNA vector. METHODS: Templates DNA of both hTERT and AR siRNA were inserted into Pgenesil vector to construct a new vector Pgenesil-hTERT-AR-shRNA by RNAi-DNA vector technology. Pgenesil-HK-shRNA, Pgenesil-hTERT-shRNA, Pgenesil-AR-shRNA and Pgenesil-hTERT-AR-shRNA vectors were transfected into prostate cancer LNCaP cells respectively. The levels of AR mRNA, apoptosis and proliferation of each cell group were determined by FQ-PCR, Annexin V method and MTT. RESULTS: The level of hTERT mRNA of control group cells and cells transfected by Pgenesil-HK-shRNA, Pgenesil-hTERT-shRNA, Pgenesil-AR-shRNA and Pgenesil-hTERT-AR-shRNA was (1.51 +/- 0.08) x 10(8), (7.32 +/- 0.43) x 10(7), (2.94 +/- 0.15) x 10(6), (4.45 +/- 0.25) x 10(7) and (3.17 +/- 0.18) x 10(6) (copies/ml) respectively. The level of AR mRNA of control cell groups and cells transfected by Pgenesil-HK-shRNA, Pgenesil-hTERT-shRNA, Pgenesil-AR-shRNA and Pgenesil-hTERT-AR-shRNA was (1.92 +/- 0.11) x 10(5), (6.47 +/- 0.32) x 10(5), (3.70 +/- 0.24) x 10(4), (1.22 +/- 0.06) x 10(4) and (7.21 +/- 0.41) x 10(3) (copies/ml) respectively. These data indicate that the expression of hTERT or AR gene could be significantly inhibited by Pgenesil-hTERT-shRNA or Pgenesil-AR-shRNA while Pgenesil-hTERT-AR-shRNA could simultaneously inhibit both hTERT and AR gene expression. The apoptosis rate and the inhibition rate of cell growth of Pgenesil-hTERT-AR-shRNA group were significantly higher than those of Pgenesil-hTERT-shRNA group or Pgenesil-AR-shRNA group (P < 0.05). CONCLUSION: It is feasible to inhibit both hTERT and AR gene expression simultaneously by single shRNA vector. It will be a new research strategy of gene therapy for prostate cancer.