Research On Neurocognitive Function And TCM Diagnosis And Treatment Of Turner Syndrom

OBJECTIVES;Neurocognitive developmental abnormalities in Turner.syndrome(TS)primarily manifest as nonverbal cognitive dysfunctions in specific learning disabilities,encompassing attention,working memory,cognitive flexibility,visuospatial skills,executive functions,mathematical logic thinking,and abstract thinking.However,neurocognitive phenotypes vary according to different chromosomal karyotypes,and the corresponding mechanism research is still insufficient.The aims of this study are:1.To investigate the differences in neurocognitive phenotypes between TS patients and healthy controls,and to explore the correlations between brain structural and functional changes and behavioral differences through brain imaging analysis;2.To delve into genetic variations,examining the genetic mechanisms underlying neurocognitive alterations in TS patients,and exploring the effects of low sex hormone expression on brain cognition and behavior during development;3.To preliminarily probe the mechanisms of Traditional Chinese Medicine treatments in ameliorating developmental brain cognitive impairments under low estrogen expression using animal experimentation.METHODS:This study was divided into three parts:1.Study 1 is a case-control study,including 30 TS girls aged 6-16 years and 15 age-and sex-matched healthy controls.Baseline information of all participants was collected,including demographics,maternal pregnancy status,family background,physical examinations,and basic laboratory tests.Intelligence,behavioral measures and neuroimaging data were collected for all subjects,with cognitive and behavioral evaluations primarily involving:cognitive language abilities,socio-emotional capabilities,motor skills,and lateralization-related measures.Neuroimaging indicators chiefly included structural MRI(gray matter volume),diffusion MRI(white matter connectivity),and resting-state functional MRI(Amplitude of low-frequency fluctuations,ALFF;Regional homogeneity,ReHo).By employing between-group comparisons and correlation analyses,differences in various indices between TS patients(45,X group and non-45,X group)and healthy controls were examined,and the impact of brain structural and functional changes on behavioral cognitive phenotypes was explored.2.Study 2 performed whole-exome sequencing assays on all study subjects,a comprehensive assessment of gonadal development,collection of data on high-frequency mutation genes,sex hormone levels(Estradiol,E2;Follicle stimulating hormone,FSH;Luteinizing hormone,LH),and uterine length,and between-group comparisons.Simultaneously,correlation analyses with neurodevelopmental indices form Study 1 were conducted to investigate the interaction relationship between gene mutations and sex hormone levels on neurocognitive development in TS patients.3.Study 3 established ovariectomized and sham-operated mouse models.Ovariectomized mice were randomly divided into four groups,treated with distilled water,low-dose estrogen,low-dose Jin Kui Shen Qi Wan,or high-dose Jin Kui Shen Qi Wan,respectively.After four weeks of gavage,all mice undergo the Morris water maze experiment,followed by six days of spatial learning and memory training,and between-group differences were compared.Tissue samples were collected,and sex hormone levels and hippocampal acetylcholinesterase were assessed in each group.HE staining was used to observe hippocampal neuronal morphology,and between-group comparisons were performed.RESULTS:1.In Study 1,we found that:(1)In behavioral cognitive assessments,TS patients demonstrated lower scores in total IQ,analogy test,matrix reasoning scores,trail making test,dotted number measure responses,mental rotation,visuospatial processing ability,fine motor skills,and bimanual coordinated ability(P<0.05);(2)In brain MRI,structural MRI revealed that TS patients had smaller gray matter volumes in total cerebral gray matter,white matter,precentral gyrus,postcentral gyrus,parahippocampal gyrus,hippocampus,temporal lobe,occipital lobe,and frontal lobe compared to healthy controls(P<0.05),whereas the gray matter volume in the left and right parieto-occipital junctions was larger(P<0.05).In diffusion MRI,TS patients exhibited lower fractional anisotropy scores in certain parietal and temporal lobe white matter regions compared to healthy controls(P<0.05).In functional MRI,TS patients had higher regional homogeneity in the cerebellum,parahippocampal gyrus,thalamus,and anterior cingulate gyrus compared to controls,while ALFF values showed no significant differences;(3)In correlation analyses,we found a positive correlation between the participants’ total IQ scores and gray matter volumes in total and in major brain regions(P<0.05).Numerosity test scores also exhibited a positive correlation with total gray matter volume and gray matter volumes in the hippocampus,temporal lobe,occipital lobe,and frontal lobe(P<0.05).Meanwhile,Purdue Pegboard Test scores showed a significant negative correlation with total gray matter volume and gray matter volume in the temporal lobe.Total IQ scores,Numerosity test scores,and left inferior parietal to inferior parietal white matter region fractional anisotropy indices demonstrated a positive correlation(P<0.05),while trail-making test scores,Purdue Pegboard Test scores,and mental rotation task scores showed a negative correlation(P<0.05).Total IQ scores,Numerosity test scores,and right inferior temporal gyrus white matter region fractional anisotropy indices exhibited a positive correlation(P<0.05),while Purdue Pegboard Test scores and mental rotation task scores displayed a negative correlation(P<0.05).The average whole-brain ReHo value was negatively correlated with Purdue Pegboard total scores(P<0.05)but showed no significant correlation with other cognitive assessments.Meanwhile,no correlation was found between total IQ scores and ReHo values in various brain regions.Trail-making test scores showed a positive correlation with ReHo values in the parahippocampal gyrus,thalamus,and anterior cingulate gyrus,while Numerosity test scores displayed a negative correlation,and Purdue Pegboard Test scores revealed a positive correlation,all with statistical significance(P<0.05).2.In Study 2,we found that autosomal gene mutations in DRD4,CHID1,MUC6,DOCK8,CD 151,ODF3L2,and GZMM were correlated with the neuroimaging and behavioral indicators in Study 1.Specifically,DRD4 gene mutation frequency was negatively correlated with ReHo values in the anterior cingulate gyrus,CHID1 gene mutation frequency was negatively correlated with matrix reasoning scores,MUC6 gene mutation frequency was negatively correlated with total IQ scores and matrix reasoning scores,and DOCK8 gene was negatively correlated with white matter volume.Additionally,CD151 gene was negatively correlated with gray matter volume in the occipital lobe,and ODF3L2 gene was negatively correlated with ReHo values in the cerebellum,parahippocampal gyrus,thalamus,and anterior cingulate gyrus.At the same time,we found that FSH was negatively correlated with total gray matter volume,white matter volume,hippocampal gray matter volume,and gray matter volumes in the temporal and occipital lobes.LH was negatively correlated with total gray matter volume,hippocampal gray matter volume,and gray matter volumes in the temporal and occipital lobes,and positively correlated with mental rotation scores.FSH/LH ratio was positively correlated with ReHo values in the parahippocampal gyrus,thalamus,and anterior cingulate gyrus.E2 was negatively correlated with ReHo values in the thalamus and anterior cingulate gyrus.Furthermore,there were no interaction effects between sex hormone level-related measurements and gene mutation frequencies on neurocognitive phenotypes.In Study 3,after 6 days of training,mice in each group demonstrated differences in regional paths in the water maze experiment(P<0.05),with the high-dose Jin Gui Shen Qi Wan treatment group performed better than the control group.The high-dose Jin Gui Shen Qi Wan treatment group had higher E2 and AchE levels than the control group and low-dose Jin Gui Shen Qi Wan group,while FSH and LH levels were lower.At the same time,there was no difference in E2,FSH,and LH measurements between the high-dose treatment group and the estrogen treatment group(P>0.05).In terms of hippocampal pathological changes,the highdose traditional Chinese medicine group significantly improved the proliferation of hippocampal neurons in ovariectomized mice.CONCLUSIONS:1.Turner syndrome patients have lower total IQ scores than agematched healthy girls,and the main behavioral and cognitive phenotypes include impaired scores in trail-making tests,dot counting tasks,mental rotation,visuospatial processing ability,fine motor,and bimanual coordination.Abnormalities in gray matter volume,white matter connectivity,and local consistency in various brain regions in TS patients may serve as the pathological basis for these behavioral and cognitive phenotypes;2.In addition to the loss of the X chromosome,autosomal gene mutations and low expression of sex hormones both have an impact on the neurocognitive development of TS,and there is no interaction between the two factors;3.Jin Gui Shen Qi Wan,a traditional Chinese medicine based on the principle of"nourishing the kidney",not only increases the sex hormone levels in ovariectomized mice but also promotes the proliferation of hippocampal neurons.