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ObjectiveTo explore the differences existing in the auditory mismatch negativity （MMN） amplitude and latency between children with attention deficit hyperactivity disorder （ADHD） and normal children， and to probe into the significance of MMN latency and amplitude for assessing the auditory perception and attention level in ADHD children and normal children. MethodsOn December 1， 2022， a systematic search was performed in PubMed， Embase， Cochrane Library， China National knowledge Infrastructure （CNKI）， Wanfang Data Knowledge Service Platform and VIP databases to identify all well qualified literature focusing on MMN of ADHD children， then the valid data relevant to MMN amplitude and latency were extracted. The Newcastle-Ottawa Scale （NOS） was used to assess the quality of the included studies， and Stata 20.0 was employed for Meta-analysis. ResultsA total of 9 qualified studies comparing ADHD children （n=170） against healthy controls （n=159） were finally included. Among the included literature， there were 18 matched pairs of MMN amplitude data and 10 matched pairs of MMN latency data at different recording sites. Meta-analysis denoted that ADHD group resulted in potentials of slightly lower MMN amplitude （WMD=-0.334， 95% CI： -1.426~0.758， P=0.549） and notably longer MMN latency （WMD=14.768， 95% CI： 4.660~24.876， P=0.004） compared to control group， and the Bgger's funnel plot did not reveal any publication bias. ConclusionCompared with healthy controls， ADHD children have longer MMN latency， suggesting that the auditory perception and attention level of ADHD children may be reduced.
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Objective:To establish Crx-iCreERT2 fluorescent reporter human embryonic stem cell lines using CRISPR/Cas9 technology and 3D retinal organoid culture.Methods:The target site sequence of H9 cell line was verified by polymerase chain reaction (PCR). SgRNAs were designed by CRISPR/Cas9 technique and their activity was detected.The most optimal sgRNA was selected according to the factors such as activity and specificity.After identification of the target vectors by restriction enzyme and sequencing, the target vectors were transferred to the H9 cell line by electroporation.P2A-tdTomato-P2A-iCreERT2 was inserted between Exon4 and 3’-untranslated region of hES-ZLM-001 gene.Knockin positive clones were obtained after drug treatment, enrichment of positive clones.Primers were designed to perform PCR on the target region, and homozygous de-resistant knockin positive cell clones were selected according to the sequencing results and peaks.The 1-A07 cell line was cultured, and then flow cytometry for the proportion of OCT4 positive cells, immunofluorescence for three stem cell molecular markers including SOX2, NANOG, SSEA4, karyotype analysis were carried out to confirm whether the 1-A07 cell line could be used for further experiments.Retinal organoids were obtained by three-dimensional (3D) culture technology and the expression of molecular markers was detected by immunofluorescence at different developmental stages of retinal organoids. Results:The target site sequence of H9 cell line was consistent with that given by Genebank and Ensembl.Sixteen sgRNAs were designed according to the target site sequence of H9 cell line, and finally sgRNA8 and sgRNA12 were selected.The sgRNAs and recombinant plasmids were transfected into the H9 cell line by electroporation, and four homozygous de-resistant knockin positive cell clones were obtained by PCR.Crx-iCreERT2 fluorescent reporter human embryonic stem cell lines were successfully obtained.In 1-A07 cell line, the proportion of OCT4 positive cells was about 98.7% by flow cytometry, and the expression of three stem cell markers was positive by immunofluorescence, and the karyotype was normal 46, XX.The results showed that the 1-A07 cell line could be used for further experiments.The Crx-iCreERT2 fluorescent reporter human embryonic stem cell lines were differentiated into tdTomato positive retinal organoids by 3D culture technology.BRN3A positive ganglion cells, CALBINDIN positive horizontal cells and CHAT positive amacrine cells appeared on day 30 of differentiation.RECOVERIN positive photoreceptors arose on day 45 of differentiation.PKCα positive bipolar cells presented on day 90 of differentiation.Ganglion cells were shown in the deep layer of retinal organoids, and horizontal cells, amacrine cells and bipolar cells in the middle layer, and photoreceptors in the top layer.Conclusions:Crx-iCreERT2 fluorescent reporter human embryonic stem cell lines are successfully established and can be differentiated into retinal organoids that express tdTomato red fluorescence through 3D culture technology.Those retinal organoids contain the same types of neurons as normal human retinas, and follow a certain temporal and spatial developmental sequence similar to the developmental rules of normal human retinas.Crx-iCreERT2 fluorescent reporter human embryonic stem cell line is a powerful tool for researching retinal development and diseases and can be applied in treatments for blindness.
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The current status and research progress of sanitation of public articles and appliances were described on the basis of the recent literature reviews.The researches on methods for disinfection of public articles and appliances showed that the most efficient method for disinfection of haircut appliance was ultraviolet radiation disinfection.The disinfetion efficiencies of different methods for disinfection of tea sets,towels,etc,the recent new reports on the selection of sanitary parameters and sampling methods of sanitary standard for public articles were introduced also.
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Objective To understand the efficiency of defluoridation facilities used in drinking water defluoridation in the rural high fluoride areas in Tianjin and the sanitary characteristics of treated drinking water. Methods The raw water samples and treated water samples were collected from 4 water stations where the electrodialysis technics was applied and 6 water stations where the reverse osmosis technics was applied and the facilities had been operated for 2-3 years in a county, as well as 3 water stations applied activated aluminium oxide adsorption technics and 2 water stations applied nanofiltration membrane technics in the other counties in Tianjin in 2002-2004. The pH value,turbidity, total dissolved solids, total hardness, fluoride, sulfate, aluminium, in all water samples were determined and evaluated according to the Sanitary Standard for Drinking Water(2001). Results The treated water samples from electrodialysis and reverse osmosis defluoridation facilities showed lower fluoride level(below 1.0 mg/L with fluoride removal rates of 90.01% and 75.32%),and lower total dissolved solids and total hardness levels with removal rates of 75.83%-93.48%. The treated water samples from nanofiltration membrane defluoridation facilities also showed lower fluoride level(below 1.0 mg/L with fluoride removal rates of 75.16%,and lower total dissolved solids and total hardness levels with removal rates of 64.90% and 77.22% respectively. Conclusion The investigated electrodialysis, reverse osmosis and nanofiltration membrane defluoridation facilities show a reliable efficiency and are suitable for application of removal of fluoride in drinking water with different levels of total dissolved solids in the rural high fluoride areas. The effects of defluoridated water produced by reverse osmosis defluoridation facilities on human health need to be studied further more.