Analysis of NSD1 gene variant in a child with autism spectrum disorder in conjunct with congenital heart disease / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical characteristics and genetic basis of a child with autism spectrum disorder (ASD) in conjunct with congenital heart disease (CHD).@*METHODS@#A child who was hospitalized at the Third People's Hospital of Chengdu on April 13, 2021 was selected as the study subject. Clinical data of the child were collected. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). A GTX genetic analysis system was used to analyze the WES data and screen candidate variants for ASD. Candidate variant was verified by Sanger sequencing and bioinformatics analysis. Real-time fluorescent quantitative PCR (qPCR) was carried out to compare the expression of mRNA of the NSD1 gene between this child and 3 healthy controls and 5 other children with ASD.@*RESULTS@#The patient, an 8-year-old male, has manifested with ASD, mental retardation and CHD. WES analysis revealed that he has harbored a heterozygous c.3385+2T>C variant in the NSD1 gene, which may affect the function of its protein product. Sanger sequencing showed that neither of his parent has carried the same variant. By bioinformatic analysis, the variant has not been recorded in the ESP, 1000 Genomes and ExAC databases. Analysis with Mutation Taster online software indicated it to be disease causing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic. By qPCR analysis, the expression level of mRNA of the NSD1 gene in this child and 5 other children with ASD was significantly lower than that of the healthy controls (P < 0.001).@*CONCLUSION@#The c.3385+2T>C variant of the NSD1 gene can significantly reduce its expression, which may predispose to ASD. Above finding has enriched the mutational spectrum the NSD1 gene.

Analysis of SIK3 gene variation in a boy with autism spectrum disorder complicated with epilepsy / 中华医学遗传学杂志

OBJECTIVE@#To study the genetic variants of a child with Autism Spectrum Disorder (ASD) combined with epilepsy, and explore its possible pathogenic mechanism.@*METHODS@#Clinical data of the child were collected and evaluated, whole-exome sequencing (WES) technology was used to explore the genetic variants sites of the child and his parents and candidate genes were filtered out. Sanger sequencing were performed to verify the variants identified by WES and PolyPhen2 was utilized to predict the function of these variants. qPCR was carry out to determine the expression of the variant gene.@*RESULTS@#The proband carried a compound heterozygous mutation in the SIK3 gene (Chr11 q23.3, NM_025164.6), which contains a missense mutation c.1295A>G (p.N432S) inherited from the father and a deletion [c.2389_2391del(p.797del)] inherited from the mother. Both mutation sites are highly conservative, and PolyPhen2 predicted (c.1295A>G [p.N432S]) to be harmful. Compared to the mother, expression of SIK3in mRNA level in the peripheral blood of the proband and his father were both significantly decreased; compared to normal child, SIK3 expression in the peripheral blood of the proband and two other children with ASD were all decreased significantly too. In addition, studies on mice found that Sik3 gene has a marked higher level of expression in the brain.@*CONCLUSION@#The SIK3 gene variants may probably be associated with ASD. The detailed mechanism needs to be studied further, which may involve lipid metabolism dysfunction in the brain.

Application of random forests regression in impact of family early disadvantaged risks on internalization problems in adolescents / 中华行为医学与脑科学杂志

Objective:To investigate the relationship between the type of exposure to early family disadvantaged risks and internalization problems in adolescents.Methods:Totally 746 adolescents were assessed with family disadvantaged risk items and Chinese version of Achenbach youth self-report (YSR-CV). The Welch's ANOVA test and post-hoc test were used to compare the scores of three different risk factors on internalization problems.The independent sample t-test was adopted to compare the scores of internalization problems between experienced 5 or more kinds of risk factors and less than 5.Pearson correlation analysis was used to test the correlation between the number of family risk factors experienced and the scores of internalization problems.Random forest regression analysis was used to test the variable importance(VI) of the internalization problem. Results:The differences in anxiety/depression and withdrawal scores between higher or lower-level risks group were significant ( P<0.05). The scores of adolescents with five or more adverse experiences on anxiety/depression(7.7±3.5), withdrawal (8.8±4.0) and physical problems(4.1±3.7) were higher than those with fewer than five risk factors(5.5±3.8), (6.7±3.4), (2.6±3.6). The cumulative family disadvantaged risk items was positively correlated with anxiety/depression( r=0.29, P<0.01), withdrawal( r=0.29, P<0.01), and physical symptoms ( r=0.26, P<0.01). The most important factor associated with anxiety/depression(VI=0.84, P=0.002; VI=0.56, P=0.022), withdrawal(VI=0.58, P=0.013; VI=0.89, P=0.001), and physical symptoms was marital relationship of parents and health status of family members. Conclusion:The parents’ marital relationship and health status of family members are the specific factors that influence the internalization problems, and the higher the level of risk adolescents experience, the more likely they are to develop internalization problems.

Effect of aspirin on microglia activation induced by Poly-IC and its regulatory mechanism / 中华行为医学与脑科学杂志

Objective:To study whether aspirin has inhibitory effect on microglia activation induced by Poly-IC and its mechanism.Methods:Microglia cell line BV2 were cultured in vitro to establish a Poly-IC stimulation-induced microglia cell immune activation model. The experiment groups were divided into control group (no treatment), model group (Poly-IC 10 μg/ml), high dose aspirin group (1 mmol/L aspirin), low dose aspirin group (0.1 mmol/L aspirin), high dose aspirin pretreatment group (Poly-IC 10 μg/ml + 1 mmol/L aspirin) and low dose aspirin pretreatment group (Poly-IC 10 μg/ml + 0.1 mmol/L aspirin). The phagocytosis ability of microglia cells, reactive oxygen species (ROS) and Iba1 protein expression were detected by using immunofluorescence method. The expression of the inflammatory cytokines Il-1β, Il-6, Il-10, TNF-α and cox-2 mRNA in microglia cells were detected by real-time quantitative PCR (RT-qPCR).Results:Compared with the control group, the morphology of microglia cells in model group changed significantly, and the phagocytosis ability and production of reactive oxygen species (ROS) increased. At the meantime, the expression of Iba1 protein was strongly decreased. In the model group, The mRNA expressions of IL-1β(20.55±1.92), IL-6 (63.98±7.83), TNF-α (16.84±3.19), COX-2 (6.78±0.42) were higher than IL-1β(1.01±0.14), IL-6 (0.95±0.17), TNF-α (1.22±0.38), COX-2 (0.87±0.11) in the control group. (Il-1β ( t=26.14), Il-6 ( t=10.22), TNF-α ( t=17.06) and COX-2 ( t=37.07), all P<0.01). In the aspirin pretreatment group, the phagocytic ability of microglia cells was inhibited compared with the model group, and the production of reactive oxygen species (ROS) reduced. The expression of Iba1 protein was also partly recovered. Meanwhile, the effect of the high aspirin dose pretreatment group on pro-inflammatory factors IL-1β(9.95±0.52), IL-6 (39.64±6.89), TNF-α(1.57±0.42), COX-2 (2.47±0.14)were lower than those in the model group significantly.(IL-1β: t=14.18, IL-6: t=3.69, TNF-α: t=16.68, COX-2: t=27.03, all P<0.01). Conclusion:Aspirin has an inhibitory effect on microglial activation induced by Poly-IC, which may be related with inhibiting the expression of inflammatory factors.

ETA-mediated anti-TNF-α therapy ameliorates the phenotype of PCOS model induced by letrozole.

Chronic inflammation is a typical characteristic of polycystic ovary syndrome (PCOS), in which, tumor necrosis factor (TNF)-α plays an important role. We investigated whether anti-TNF-α therapy can alleviate the core phenotypes of PCOS. In pubertal female Wistar rats, release pellets of letrozole (LET) were administered continuously for 90 days to induce PCOS-like phenotypes, followed by treatment with etanercept (ETA), a TNF-α inhibitor. ETA significantly inhibited increases in body weight and androgen, TNF-α, and MCP-1 levels, excessive recruitment of lipid droplets, altered levels of pre-adipose differentiation markers, and abnormal development of follicles. In addition, TNF-α and testosterone (T) levels in the rat sera were significantly positively correlated. Further experiments were performed to investigate the relationship between TNF-α and androgen. Persistent exposure of the RAW 264.7 cell line to low doses of testosterone significantly enhanced TNF-α expression and activated the NF-κB signaling pathway, which were blocked by ETA. Furthermore, treatment with TNF-α promoted the production of testosterone in KGN granulosa cells by reducing CYP19A1 expression, whereas ETA treatment blocked this process. In conclusion, anti-TNF-α therapy with ETA may be an efficient method to alleviate PCOS, whose underlying mechanism may be associated with its ability to reduce excessive androgen levels.

CFTR Deletion in Mouse Testis Induces VDAC1 Mediated Inflammatory Pathway Critical for Spermatogenesis.

Cystic fibrosis is the most common genetic disease among Caucasians and affects tissues including lung, pancreas and reproductive tracts. It has been shown that Endoplasmic Reticulum (ER) stress and heat shock response are two major deregulated functional modules related to CFTR dysfunction. To identify the impact of CFTR deletion during spermatogenesis, we examined the expression of spermiogenesis-related genes in the testis of CFTR mutant mice (CF mice). We confirmed expression changes of MSY2, a germ cell specific RNA binding protein, resulting from deletion of CFTR in testis. Furthermore, real time PCR and Western blot results showed that an inflammatory response was activated in CF mice testis, as reflected by the altered expression of cytokines. We demonstrate for the first time that expression of MSY2 is decreased in CF mice. Our results suggest that CFTR deletion in testis influences inflammatory responses and these features are likely to be due to the unique environment of the seminiferous tubule during the spermatogenesis process. The current study also suggests avenues to understand the pathophysiology of CFTR during spermatogenesis and provides targets for the possible treatment of CFTR-related infertility.

Association of polymorphisms of miRNA biogenesis related genes DICER and DROSHA with azoospermia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of polymorphisms of miRNA biogenesis related genes DICER and DROSHA with azoospermia.</p><p><b>METHODS</b>For 330 patients with primary azoospermia and 282 fertile male controls, single nucleotide polymorphisms (SNPs) of DICER rs3742330 and DROSHA rs10719 were determined with a restriction fragment length polymorphism (RFLP) method.</p><p><b>RESULTS</b>For the SNP rs3742330, the frequency of A allele was higher among azoospermia patients compared with the controls (72.0% vs.64.4%, P=0.004), and so was the frequency of AA genotype (53.0% vs. 41.8%, P=0.027, OR=1.829, 95%CI: 1.071-3.124). On the other hand, the allelic and genotypic frequencies of rs10719 did not differ between the two groups (all P > 0.05).</p><p><b>CONCLUSION</b>Polymorphisms of rs3742330 of the DICER gene, particularly the AA genotype, may be associated with azoospermia.</p>

Analysis of DIAPH3 gene mutation in a boy with autism spectrum disorder / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the clinical manifestations and gene mutation of a 6 year old boy with autism spectrum disorders (ASD).</p><p><b>METHODS</b>Peripheral blood of the boy and his parents were subjected to genetic testing.</p><p><b>RESULTS</b>The patient was diagnosed with typical autism. Exome sequencing has identified mutations of four candidate genes, namely TUT1, DIAPH3, REELIN and SETD2, which were confirmed with Sanger sequencing. Analysis of family members confirmed that the missense mutations of DIAPH3 and SETD2 genes were of de novo origin.</p><p><b>CONCLUSION</b>Missense mutations of DIAPH3 and SETD2 genes may have contributed to the risk of ASD. Disrupted neurogenesis associated with such mutations may have been the underlying mechanism for ASD.</p>

A method for bleeding detection in endoscopy images using SVM / 中国医疗器械杂志

Because the huge number of images of the digestive tract by Wireless Capsule Endoscopy (WCE) are left to the medical personnels detected by their eyes, huge burden leaves to doctors. This article provides a classification of method based on SVM (Support Vector Machine) for the capsule endoscopy bleeding intelligent recognition. We created a new kind of feature parameter, and the experiment result can reach 83% specificity and 94% sensitivity.

Prevalence of psychiatric disorders among children and adolescents in northeast China.

BACKGROUND: To describe the prevalence of DSM-IV disorders and comorbidity in a large school-based sample of 6-17 year old children and adolescents in northeast China. METHODS: A two-phase cross-sectional study was conducted on 9,806 children. During the screening phase, 8848 children (90.23%) and their mothers and teachers were interviewed using the Strengths and Difficulties Questionnaire (SDQ). During the diagnostic phase, 1129 children with a positive SDQ and 804 randomly selected children with a negative SDQ (11%), and their mothers and teachers, were interviewed using the Development and Well-Being Assessment (DAWBA). RESULTS: The overall prevalence of DSM-IV disorders was 9.49% (95% CI = 8.10-11.10%). Anxiety disorders were the most common (6.06%, 95% CI = 4.92-7.40), followed by depression (1.32%, 95% CI = 0.91-1.92%), oppositional defiant disorder (1.21%, 95%CI = 0.77-1.87) and attention-deficit hyperactivity disorder (0.84%, 95% CI = 0.52-1.36%). Of the 805 children with a psychiatric disorder, 15.2% had two or more comorbid disorders. CONCLUSIONS: Approximately one in ten Chinese school children has psychiatric disorders involving a level of distress or social impairment likely to warrant treatment. Prevention, early identification and treatment of these disorders are urgently needed and pose a serious challenge in China.

Edaravone protects H9 c2 cells against doxorubicin-induced cardiotoxicity / 中国药理学通报

Aim To explore whether edaravone (EDA), a novel free radical scavenger, protects H9c2 cardiac cells against doxorubicin ( DOX )-induced car-diotoxicity. Methods H9c2 cells were treated with 5μmol·L-1 DOX to establish a model of DOX cardio-toxicity. Cell viability was examined by cell counter kit ( CCK-8 ) . Changes in morphology and amount of ap-optotic cells were detected by Hoechst 33258 staining;intracellular level of reactive oxygen species ( ROS ) was measured by DCFH-DA staining and photofluorog-raphy;mitochondrial membrane potential ( MMP) was observed by rhodamine 123 ( RH123 ) staining and photoflurograph; the expression level of caspase-3 was determined by Western blot assay. Results Pretreat-ment of H9 c2 cells with 20 , 40 and 80 μmol · L-1 EDA for 60 min markedly inhibited cytotoxicity in-duced by 5 μmol · L-1 DOX, respectively, as evi-denced by an increase in cell viability. The protective effect induced by 40 μmol · L-1 EDA was maximal. Pretreatment of H9 c2 cells with 40 μmol · L-1 EDA for 30 , 60 , 90 and 120 min significantly attenuated DOX-induced cytotoxicity, respectively, having a max-imal protection at 60 min. Furthermore, pretreatment of H9 c2 cells with 40 μmol · L-1 EDA for 60 min be-fore exposure to 5 μmol · L-1 DOX for 24 h obviously reduced cardiac injuries, as evidenced by decreases in the DOX-induced intracellular ROS generation, num-ber of apoptotic cells, and expression of cleaved caspase-3, as well as loss of MMP. Conclusions EDA can protect H9 c2 cardiac cells against DOX-in-duced cardiotoxicity, this protection may be associated with inhibition of ROS production and preservation of MMP.

Expression pattern of congenital chloride diarrhea pathogenic gene Slc26a3 in the reproductive tract of male rodents / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To determine the expression pattern of Slc26a3 gene in reproductive tract of male rodents to clarify whether the expression pattern is related to the subfertility observed in congenital chloride diarrhea (CLD) disease.</p><p><b>METHODS</b>The expression of Slc26a3 in mouse and rat epididymis has been studied with immunohistochemistry and Western blotting. Its developmental expression pattern in rat testis was detected by Western blotting, while both of immunofluorescence and Western blotting were used to localize the expression of Slc26a3 in mouse sperms. The potential change of Slc26a3 expression in CFTR (cystic fibrosis transmembrane conductance regulator) knockout mice and CFTR mutant mice was also detected with Western blotting.</p><p><b>RESULTS</b>The expression level of Slc26a3 gradually decreased along epididymis from its caput to corpus, then to its cauda part. This gradually decreasing expression pattern was also found in rat testis during development. Slc26a3 was localized mainly on the trunk of mouse sperm tail. In the testis and epididymis of CFTR knockout mice and CFTR mutant mice, no significant change of Slc26a3 expression was found.</p><p><b>CONCLUSION</b>Slc26a3 is expressed in male reproductive tract, and its expression pattern is related to the function. Thus, the subfertility observed in CLD disease may be related to the important role of SLC26A3 in acid-base regulation of epididymis.</p>

Comparison of Testing Results for Coliform Bacteria and Escherichia Coli in Oral Drugs / 中成药

Objective: To compare with coliform bateria and Escherichia coli testing results in oral drugs. Methods: 24 species Chinese herb medicines and 35 kinds of Chinese traditional patent medicines in 68 batches were examined by using the method of coliform bacteria and escherichis coli test.Results: There were significant differences. Conclusion: The result showed that using the coliform bacterias as the hygienic indicative bacteria was significant.