Autism Spectrum Disorders (ASDs) are reported as a group of neurodevelopmental disorders. The structural changes of brain regions including the hippocampus were widely reported in autistic patients and mouse models with dysfunction of ASD risk genes, but the underlying mechanisms are not fully understood. Here, we report that deletion of Trio, a high-susceptibility gene of ASDs, causes a postnatal dentate gyrus (DG) hypoplasia with a zigzagged suprapyramidal blade, and the Trio-deficient mice display autism-like behaviors. The impaired morphogenesis of DG is mainly caused by disturbing the postnatal distribution of postmitotic granule cells (GCs), which further results in a migration deficit of neural progenitors. Furthermore, we reveal that Trio plays different roles in various excitatory neural cells by spatial transcriptomic sequencing, especially the role of regulating the migration of postmitotic GCs. In summary, our findings provide evidence of cellular mechanisms that Trio is involved in postnatal DG morphogenesis.
Animals ; Dentate Gyrus/metabolism* ; Mice ; Morphogenesis/physiology* ; Neurons/pathology* ; Cell Movement ; Mice, Inbred C57BL ; Autism Spectrum Disorder/pathology* ; Mice, Knockout ; Neural Stem Cells ; Male ; Neurogenesis ; Autistic Disorder/genetics*

Objective:To investigate the patterns of mutation evolution in patients with acute myeloid leukemia (AML) during treatment and the possible clinical significances.Methods:A retrospective case series study was conducted. A total of 103 AML patients who were hospitalized at the Affiliated Yuebei People's Hospital of Medical College of Shantou University from November 2019 to August 2021 and underwent high-throughput next-generation sequencing (NGS) technology to detect the mutations of 128 AML-related genes in bone marrow samples were selected. Based on the NGS results, the somatic gene mutations in samples of patients collected at initial diagnosis (73 cases), complete remission (CR) (30 cases), non-remission (NR) (23 cases), and recurrence (12 cases) were analyzed, and the targeted drugs involved in the gene mutations detected in NR and recurrence samples were summarized.Results:The median age [ M ( Q1, Q3)] of onset for 103 patients was 58 (48, 66) years, including 64 males (61%) and 39 females (39%); 86 cases (83%) were primary AML, and 17 cases (17%) were secondary AML; at the initial diagnosis, 51 cases (50%) had normal karyotypes, 34 cases (33%) had abnormalities, and 18 cases (17.5%) were unknown. Compared with the CR samples, the mutation frequencies of FLT3 [29% (21/73) vs. 3% (1/30)], NPM1 [27% (20/73) vs. 3% (1/30)], NRAS [22% (16/73) vs. 3% (1/30)], and IDH2 [14% (10/73) vs. 0 (0/30)] were all higher in the initial diagnosis samples, and the differences were statistically significant (all P < 0.05); compared with the initial diagnosis sample, the median number of gene mutations in each CR sample was lower [4 (2, 5) vs. 7 (5, 9)], and the difference was statistically significant ( P < 0.001). However, there was no statistically significant difference in the median number of gene mutations in each patient between the initial diagnosis samples and the NR samples, the initial diagnosis samples and the recurrence samples, and the NR samples and the recurrence samples (all P > 0.05). Analysis of 14 patients with NGS data at initial diagnosis and CR showed that the same gene mutations could be detected at initial diagnosis and CR, such as DNAH23 (3 cases), USH2A (3 cases), etc; partial gene mutations were detected at initial diagnosis but were not detected at CR, including NRAS (5 cases), FLT3 (3 cases), ANKRD26 (3 cases), NPM1 (3 cases), ETV6 (3 cases), etc; ARID1B (1 case) and DNMT3A (1 case) were negative for mutations at initial diagnosis but positive upon reaching CR. Analysis of 14 patients with NGS data at initial diagnosis and NR showed that most gene mutations persisted at initial diagnosis and NR, such as DNMT3A (5 cases), NRAS (5 cases), KRAS (3 cases), RUNX1 (3 cases), etc; the mutant genes detected at initial diagnosis but not detected at NR included USH2A (2 cases), PCLO (2 cases), ATM (2 cases), FAT1 (2 cases), etc; partial gene mutations were not detected at initial diagnosis but were detected at NR, such as FAT1 (2 cases), TCF3 (2 cases), etc. Analysis of 5 patients with NGS data at CR and recurrence showed that some gene mutations were detected at both CR and recurrence, such as BCORL1 (1 case), ARID2 (1 case), SETD2 (1 case), VEGFC (1 case), etc; FLT1 (1 case) and GNAS (1 case) gene mutations were detected at CR but not detected at recurrence; at recurrence, some gene mutations that were not detected at CR were also detected, such as ANKRD26 (1 case), WT1 (1 case), etc. Among the 23 NR samples and 12 recurrence samples, the targets of drugs approved by US Food and Drug Administration or in clinical trials were detected in 14 (61%) and 5 (42%) samples respectively, including IDH1, IDH2, FLT3, KIT, KRAS, NRAS, SF3B1, U2AF1, and SRSF2. Conclusions:The number of gene mutations in AML patients during CR is significantly less than that at initial diagnosis, some gene mutations disappear when CR is achieved through treatment, but the majority of gene mutations persist during the treatment period, including NR and recurrence, suggesting that monitoring through NGS technology can help understand the evolution of gene mutations during AML treatment and discover the potential therapeutic targets.

Objective To identify mechanisms regulating disease progression in rat models of pulmonary arterial hy-pertension(PAH).Methods Rat PAH models were established using subcutaneous monocrotaline(MCT)injec-tion and the SU5416/hypoxia(SU/Hx)method.Transcriptomic sequencing of lung tissues was performed to identify gene expression and pathway alterations in PAH rats,followed by a comparative analysis with transcriptomic data of patients with idiopathic pulmonary arterial hypertension(IPAH)in NCBI database.Results Inflammatory-related genes such as CXCL9,CCL24,and SECTM1 were upregulated in both PAH rat models and IPAH patient lungs,while genes such as DGKG and DOCK9 were downregulated(P＜0.05).Pathways related to endothelial cell proliferation regulation and extracellular matrix(ECM)remodeling were significantly upregulated(P＜0.05).Conclusions The imbalance in endothelial cell proliferation and abnormal ECM remodeling may collectively contribute to PAH pathogenesis.Further exploration of these signaling pathways may provide deep in-sights for early diagnosis and targeted therapy of PAH.

Objective:To observe the effects of transcranial direct current stimulation (tDCS) on functional connectivity (FC) in language-related brain regions of patients with picture-naming dysfunction after cerebral infarction by using resting state functional magnetic resonance imaging(rs-fMRI).Methods:Twenty-eight patients with post-infarction picture-naming dysfunction were divided into an acute stage group( n=16) and a recovery stage group( n=12) according to the course of the disease, and 18 middle-aged and elderly volunteers were recruited as the normal control group.The anodic tDCS was applied on the posterior perisylvian region(PPR) of the left sylvian of the patients, 5 days a week for 2 weeks.Before and after the 2 weeks′ treatment, the rs-fMRI and Psycholinguistic Assessment of Chinese Aphasia (PACA)-picture-naming subscale were performed, and FC changes in language-related brain areas were observed. Results:After treatment, the PACA scores of patients in both acute and recovery stage groups were significantly improved after treatment( P<0.05). Compared with normal subjects, FC in multiple brain regions and particularly the Wernicke area was reduced in both cerebral hemispheres among the patient group. It was more severe in the dominant hemisphere.After the tDCS treatment, FC in both frontotemporal lobes and in the Wernicke area was significantly enhanced in both the acute and recovery groups. Further comparison showed that in the acute group FC in both temporo-occipital lobes was significantly enhanced after treatment. In the recovery group, the enhanced FC in the left temporal lobe before the treatment was significantly reduced after treatment. Conclusion:The fMRI technique can evaluate changes in brain connectivity in aphasia patients with picture-naming dysfunction after cerebral infarction accurately and non-invasively.tDCS may improve picture-naming function of stroke patients by enhancing the FC in bilateral language-related brain areas(concentrated in frontotemporal lobes) and Wernicke area.

Objective The objective of this study was to provide methodological references for the inheritance of the experience of well-known Chinese medicine doctors in the treatment of kidney disease.Methods The study collected medical case data for IgA nephropathy,diagnosed and treated by Professor Yang Nizhi's outpatient department at Guangdong Provincial Hospital of Traditional Chinese Medicine from 2010 to 2020.The data was standardized and divided into three groups:urine and blood,urine turbidity,and renal failure groups.The study utilized the FangNet platform to apply the PageRank algorithm and calculate the THScore of different subgroups of core herbs for IgA nephropathy.The distribution pattern of syndrome differentiation and corresponding herb use regulations were visualized through Python(SciPy package,Clusterheatmap package),and the study explored and verified the drug prescription through exploratory and confirmatory factor analysis based on Pearson correlation coefficient.The weighted least squares estimation mean and variance adjusted(WLSMV)and the oblique rotated GEOMIN method were used with the Mplus software.Results The study included a total of 548 treatments for 145 patients with IgA nephropathy,with heamturia group(54 cases),urine turbidity group(51 cases),and renal failure group(40 cases).Results showed 9 basic syndromes such as Qi deficiency syndrome(91.79%),blood stasis syndrome(77.01%),damp-heat syndrome(66.06%),and Yin deficiency syndrome(38.69%).There are 24 core drugs in total,23 in the urine and blood group,21 in the urine turbidity group,and 16 in the renal failure group.These drugs mainly include qi-tonifying and yang-invigorating drugs,nourishing yin and blood drugs,promoting blood circulation and removing blood stasis drugs,and clearing heat and cooling blood drugs.The regulations for the differentiation and medication of IgA nephropathy(Z-Score>0.5 and P<0.05)were as follows:Huangqi,Shan Zhu Yu,and Tusizi were commonly used in Qi deficiency syndrome;Danshen,Ze Lan,and Shan Zhu Yu were commonly used in blood stasis syndrome;Pu Gong Ying,Shi Wei,Tao Ren,and Tu Fu Ling were commonly used in damp-heat syndrome;and Mo Han Lian,Tai Zi Shen,and Nv Zhen Zi were commonly used in Yin deficiency syndrome.Through exploratory and confirmatory factor analysis,the core drug combination factors for the treatment of IgA nephropathy by Professor Yang Nizhi were obtained as follows:F1(Tusizi,Shan Zhu Yu,Huangqi);F2(White Mao Gen,Xiao Ji,Qian Cao);F3(Nv Zhen Zi,Mo Han Lian,Tai Zi Shen);and F4(Ze Lan,Tao Ren).Conclusion This study analyzed the diagnosis and treatment experience of Professor Yang Nizhi in the treatment of IgA nephropathy by grouping,defining the core syndrome of"Qi deficiency and blood stasis,damp-heat and Yin deficiency",and the core treatment methods of"tonifying Qi,promoting blood circulation,clearing heat,and nourishing Yin"using the PageRank algorithm and Mplus factor analysis.The study provided methodological references for the inheritance of the experience of famous Chinese medicine doctors and promoted the development and utilization of traditional Chinese medicine.

Experiential teaching is a practical, reflective, and situational teaching method. Based on the systematic review of literature published worldwide, this paper summarizes the characteristics and organizational forms of experiential teaching, with a focus on its current application and existing problems in teaching geriatric nursing. This review provides suggestions for the reform of teaching methods of geriatric nursing in China.

Objective:To document any effect of transcranial direct current stimulation (tDCS) on the picture naming ability of stroke survivors with aphasia.Methods:Twenty-eight aphasic stoke survivors with picture naming dysfunction were divided into an acute group (with a course of disease of <1 month) and a convalescent group (with a course of disease of 2 to 6 months). Eighteen healthy subjects well-matched for age, gender and years of education formed the healthy control group. The patient group received tDCS once a day, 5 days a week for 2 weeks. Before and after the intervention, they were assessed using the Chinese psycholinguistic aphasia assessment (PACA) instrument. The activation of speech-related brain areas in everyone was quantified using resting state functional magnetic imaging (rs-fMRI).Results:After treatment the average PACA image naming scores of both the acute and convalescent groups had improved significantly. ALFF showed significant positive activation in the patient group′s right inferior temporal gyrus and negative activation in their left posterior central gyrus, while ReHo was significantly and positively activated in the right orbital superior frontal gyrus, the left medial superior frontal gyrus, the pericalar fissure cortex and the left parietal gyrus. It was, however, significantly negatively activated in the right posterior central gyrus. In the acute stage group, the ALFF was significantly and positively activated in the right superior frontal gyrus and right angular gyrus after the treatment, while the significant positive ReHo activation was in the right direct gyrus, the right angular gyrus, the right superior frontal gyrus and the right inferior temporal gyrus. In the convalescent group after the intervention the ALFF was significantly and positively activated in the left middle occipital gyrus and the right fusiform gyrus but negatively and significantly activated in the right insula, while ReHo was significantly and positively activated in the left superior temporal gyrus and the right angular gyrus.Conclusions:tDCS can improve the image naming of aphasic stroke survivors. The compensatory activation of language function is mainly in the right hemisphere in the acute stage, but in the convalescent stage the unimpaired brain area of the left cerebral hemisphere is also activated. The long-term recovery of language functioning may be the result of synergy between the hemispheres.

Most α2-AR agonists derived from dexme-detomidine have few structure differences between them and have no selectivity for α2A/2B-AR or Gi/Gs,that can lead to the side effect of drugs.To get novel and potent α2A-AR agonists,we built the homology model for human α 2A-AR and α2B-AR to find α2A-AR agonists with higher selectivity.Compound P300-2342 and its 3 analogs sig-nificantly decreased the locomotor activity of mice(P＜0.05).Furthermore,P300-2342 and its 3 analogs inhibited the binding of[3H]rauwolscine to α 2A-AR and α 2B-AR respectively.In α2A-AR-HEK293 cells,P300-2342 decre-ased forskolinstimulatedcAMPpruductionwithoutincreas-ing cAMP pruduction,that indicated the P300-2342 acti-vating α2A-AR coupling with Gαi/o pathway without Gαs coupling.P300-2342 had no agonistic and antagonistic activities on α 2B-AR.Similar results were shown in 3 analogs of P300-2342.The docking results showed that P300-2342 formed the π-hydrogen bonds with Y394,V114 of α2A-AR,and with V93 of α2B-AR.3 analogs of P300-2342 formed several π-hydrogen bonds with V114,Y196,F390 of α 2A-AR and with V93 of α 2B-AR.We believe that these molecules can serve as leads for fur-ther optimization of α2A-AR agonists with potentially few side effects.

Objective:To investigate the characteristics of transcranial sonography (TCS) in patients with restless legs syndrome (RLS), and analyze the correlations of scores of RLS Self-rating Severity Scale by International Restless Leg Syndrome Study Group (IRLS) and TCS parameters with clinical data of these patients.Methods:Twenty-one patients with RLS admitted to the Sleep Disorder Clinic of our hospital from September 2020 to January 2021 were selected as RLS group, and 23 healthy controls at the same time period were recruited as control group. IRLS was used to evaluate the severity of patients in the RLS group, and the 14-item Hamilton anxiety rating scale (HAMA-14) and 24-item Hamilton depression rating scale (HAMD-24) were used to evaluate the anxiety and depression of subjects from the 2 groups. Pittsburgh Sleep Quality Scale (PSQI), Insomnia Severity Index (ISI) and Epworth Sleepiness Scale (ESS) were used to evaluate the sleep quality of subjects from the 2 groups. TCS was used to examine the occurrence of hypoechoic substantia nigra and raphe nucleus rupture and the width of the third ventricle in the two groups. The clinical data and TCS parameters of patients in the 2 groups were compared, and the correlations of IRLS scores and TCS parameters with clinical features of patients in the RLS group were analyzed.Results:As compared with those in the control group, the HAMA-14, HAMD-24, ISI and PSQI scores in the RLS group were statistically higher ( P<0.05). As compared with the control group, RLS group had significantly higher proportion of patients with hypoechoic substantia nigra or raphe nucleus rupture ( P<0.05). In RLS patients, the IRLS scores were positively correlated with HAMA-14, HAMD-24, and ISI scores ( P<0.05); ESS scores were negatively correlated with hypoechoic substantia nigra and width of the third ventricle ( rs=-2.005, P=0.045; r=-0.477, P=0.029); width of the third ventricle was negatively correlated with gender (male) and years of education ( rs=-0.592, P=0.005; r=-0.627, P=0.002), and positively correlated with age and course of the disease ( r=0.756, P<0.001; r=0.167, P=0.047). Conclusions:Patients with RLS are prone to anxiety, depression and sleep disorders; their TCS shows hypoechoic substantia nigra and raphe nucleus rupture. RLS severity may affect HAMA-14, HAMD-24, and ISI scores. Gender, age, years of education, course of disease, and ESS scores of RLS patients may affect TCS related parameters.

Objective:To investigate the effect of low-frequency ultrasound combined with hydroxychloroquine (HCQ) on the growth and invasion of 4T1 breast cancer in mice.Methods:4T1 cells in logarithmic growth phase were divided into 4 groups: control group, ultrasound group, HCQ group and ultrasound combined with HCQ group. Western blot was performed to detect the effects of ultrasound combined with HCQ on the protein expression levels of autophagy-related proteins LC3, p62 and matrix metalloproteinase 9 (MMP-9). Transmission electron microscopy was used to observe the formation of autophagic vesicles. The cell proliferation and cell viability were determined by EdU and CCK-8. Transwell assay was performed to detect the effect of ultrasound combined with HCQ on the invasive ability of 4T1 cells. Flow cytometry was performed to detect the effect of ultrasound combined with HCQ on the apoptosis of 4T1 cells. The transplantation tumor model of 4T1 breast cancer in BALB/c mice was constructed.The mice were randomly divided into 4 groups ( n=5 for each group), including control group, ultrasound group, HCQ group, ultrasound combined with HCQ group. The tumor volume and mice body weight were evaluated and measured in each group every 2 days. Results:The expression of LC3-Ⅱ and p62 protein levels increased in the ultrasound combined with HCQ group, and intracellular autophagosome accumulation was evident by transmission electron microscopy. In cellular experiments, compared with the other groups, the ultrasound combined with HCQ group showed stronger growth inhibition, significantly decreased cell proliferation rate, decreased expression of MMP-9, and significantly inhibited invasion (all P<0.050). In the in vivo experiments, compared with the control group, the tumor growth rate of all 3 inter vention groups of mice decreased (all P<0.050), and the ultrasound combined with HCQ group had better therapeutic effects than the ultrasound and HCQ groups. The treatment effect of ultrasound combined with HCQ group was better than that of ultrasound and HCQ groups (all P<0.001). Conclusions:The combination of low-frequency ultrasound and hydroxychloroquine could synergistically inhibit tumor cell proliferation, promote apoptosis, and significantly inhibit tumor growth in 4T1 tumor-bearing mice.