Objective To investigate the metabolite changes in the preschool children with autism spectrum disorder (ASD) using MR spectroscopy (MRS) and explore the associations between image findings and clinical variables, which may provide a noninvasive brain biochemical method for the early diagnosis and prevention of autism. Methods Twenty one cases of preschool ASD children (3-6 years old) and age-and sex-matched 20 preschool healthy controls underwent single voxel short (SVS) short TE (TE=30 ms) MRS. The absolute metabolite concentrations in the anterior cingulate cortex (ACC) , anterior middle anterior cingulate cortex (aMCC) and posterior cingulate (PCC) were quantitatively analyzed using LCModel software. Two independent sample t tests were used for analysis. The relationships between metabolite concentrations and diagnostic and statistical manual of mental disorders (DSM-IV) , childhood autism rating scale (CARS) and autism behavior checklist (ABC) were analyzed by Pearson correlation analysis. Results Compared to control subjects, ASD patients had significantly lower N-acetylaspartate (NAA) values (4.35 ± 0.80, 6.34±0.82, 8.04±0.97 mmol/L respectively) in ACC, aMCC and PCC (t=2.640, P=0.012;t=2.182, P=0.035;t=3.343, P=0.002) , had significantly lower choline (Cho) 1.32±0.22 mmol/L (t=2.905, P=0.006) and glutamine and glutamate complex (Glx) 10.02 ± 0.88 mmol/L (t=2.090, P=0.043) in PCC. Cho, total creatine (tCr) , myo-Inositol (MI) and Glx levels did not differ between groups in other aforementioned regions (P>0.05). Negative correlations between the NAA ualues in the PCC and CARS (r=-0.504, P=0.020) were detected, and no significant correlation among DSM-IV, CARS, ABC and other metabolite values (P>0.05). Condnsions The biochemical changes in the preschool children with ASD in cingulate reflect the neuronal loss, structural or functional damage and cell membrane enzyme metabolic dysfunctions, may reveal the pathological basis of ASD. These results may provide noninvasive and quantitative methods for the diagnosis and prognosis evaluation of ASD child.

Objective To explore the executive function profiles in adults with attention deficit hyperactivity disorder(ADHD)in a Chinese sample population. Methods We identified 68 outpatients fulfilling the diagnostic criteria for ADHD according to the Diagnostic and Statistical Manual of Mental Disorders-IV(DSM-IV)and 74 normal adults.All subjects completed clinical interview, Behavior Rating Inventory of Executive Function-Adult Version (BRIEF-A) and Cambridge Neuropsychological Test Automated Battery (CANTAB). Results ADHD group had higher scores in all factors of BRIEF-A questionnaire compared with normal control (P<0.01). ADHD group had a higher score in SWM strategy score [(30.84±5.58)vs.(28.80±5.85), t=2.056,P=0.042]compared with control group. Compared with control group, ADHD group had a lower score in RVP total hits [37(33,41.75) vs.3(37,48), Z=-3.184,P<0.01]. Total correct rejections[(508(497.5,518)vs. 516(503,529.5),Z=-2.229,P=0.0260)]were significantly lower,while RVP total misses were significantly higher [(17 (12.25,21)vs.12(7,20),Z=2.293,P=0.022)].ADHD-RS-IV attention deficit score was positively correlated with SWM strategy while RVP total misses were negatively correlated with RVP total hits and RVP correct rejections.Working memory score in BRIEF-A was significantly correlated with the SWM strategy,RVP total hits, RVP total correct rejections and RVP probable of hit as well as RVP total misses. Conclusion Adults with ADHD have executive functioning deficits which is correlated with ADHD core symptoms.

AIM: To study the effects of fibroblast growth factor 8 (FGF8) on directional differentiation of human dental pulp stem cells (hDPSCs) into odontoblasts and pulp tissue.METHODS: hDPSCs were isolated and cultured, and identified with flow cytometry by detecting cell surface markers of hDPSCs.FGF8 at concentration of 50 μg/L was added into the mineralization fluid to induce the differentiation of the hDPSCs.The mRNA expression of dentin sialophosphoprotein (DSPP), alkaline phosphatase (ALP), bone sialoprotein (BSP) and core-binding factor alpha 1 (Cbfa-1) in differentiated cells was detected by real-time PCR.FGF8 and mouse E11.5 dental epithelium formed restructuring cell group with hDPSCs, and then the restructuring cell group was transplanted under renal capsule membrane in nude mice for tissue culture.DNA in situ hybridization was used to identify the sources of odontoblasts and pulp cells.RESULTS: The surface markers of CD29 and CD90 showed positive in isolated hDPSCs.FGF8 induced hDPSCs to form a distinct mineralization nodule, and the expression of dentin-specific proteins, DSPP, BSP and Cbfa-1, was increased.hDPSCs were induced to differentiate into odontoblasts and pulp cells by E11.5 dental epithelium and FGF8.CONCLUSION: FGF8 can assist dental epithelium to induce directional differetiation of hDPSCs into odontoblasts and pulp cells, and formation of dentin and dental pulp cavity structure.

Objectives: The primary aim of this study is to discern the association between specific clinical parameters and low muscle mass (LMM). We endeavor to elucidate the determinants of LMM and the predictive potency of individual factors. Methods: In this retrospective cross-sectional study, we encompassed 450 older adult Chinese participants (252 males and 198 females). Muscle mass quantifications were performed using bioelectrical impedance analysis.Comprehensive data encompassing demographic details (age, sex, height, and weight) and laboratory results (complete blood count, thyroid function, liver function, and renal function) were systematically recorded. Lo gistic regression models, coupled with receiver operating characteristic curve analytics, were employed to ascertain the variables influencing LMM and to evaluate the predictive validity of each parameter on LMM. Results: Upon confounding adjustment for age, gender, body mass index (BMI), and free thyroxine (FT4) persisted as a determinant of LMM. Specifically, individuals with an FT4 exceeding 1.105 ng/dL exhibited a 1.803-fold increased propensity for LMM relative to those with FT4 values below the specified threshold. Incorporating age, gender, BMI, and FT4 in the diagnostic algorithm enhanced the precision of LMM. The results differ between men and women. In the male population, we can still observe that FT4 has a certain value in the diagnosis of LMM, but this phenomenon is not found in the female population. Conclusions Elevated FT4 concentrations, albeit within clinically accepted limits, are inversely associated with muscle mass. As such, FT4 could be postulated as a potential biomarker for LMM in geriatric individuals, especially in the male group.

OBJECTIVETo quantitatively analyze the temporal and spatial pattern of RhoA expression in injured spinal cord of adult mice.

METHODSA spinal cord transection model was established in adult mice. At 1, 3, 7, 14, 28, 56 and 112 days after the surgery, the spinal cords were dissected and cryosectioned for RhoA/NF200, RhoA/GFAP, RhoA/CNPase or RhoA/IBA1 double fluorescent immunohistochemistry to visualize RhoA expressions in the neurons, astrocytes, oligodendrocytes and microglia. The percentages as well as the immunostaining intensities of RhoA-positive cells in the parenchymal cells were quantitatively analyzed.

RESULTSRhoA was weakly expressed in a few neurons and oligodendrocytes in normal spinal cord. After spinal cord injury, the percentage of RhoA-positive cells and RhoA expression intensity in the spinal cord increased and peaked at 7 days post injury (dpi) in neurons, oligodendrocytes and astrocytes, followed by a gradual decrease till reaching a low level at 112 dpi. In the microglia, both the RhoA-positive cells and RhoA expression intensity reached the maximum at 14 dpi and maintained a high level till 112 dpi.

CONCLUSIONTraumatic spinal cord injury can upregulate RhoA expression in the neurons as well as all the glial cells in the spinal cord. RhoA expression patterns vary with post-injury time, location and among different parenchymal cells in the injured spinal cord.

Animals ; Astrocytes ; metabolism ; Female ; Mice ; Mice, Inbred Strains ; Microglia ; metabolism ; Neuroglia ; metabolism ; Neurons ; metabolism ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; metabolism ; rho GTP-Binding Proteins ; metabolism

OBJECTIVE: To explore the molecular pathogenesis for a pedigree affected with hereditary coagulation factor XII (FXII) deficiency. METHODS: Potential variant of the F12 gene was analyzed by PCR and Sanger sequencing. Expression plasmids were constructed by site-directed mutagenesis based on the wild-type and transiently transfected into 293T cells. FXII:C and FXII:Ag of the expression products were determined in the supernatant and cell lysate. Western blotting was used to verify the identify of the protein. RESULTS: Gene sequencing revealed that the proband has carried 46TT genetype and heterozygous p.Glu502Lys variants in exon 13, and a heterozygous p.Gly542Ser variant in exon 14 of the F12 gene. Transfection experiment suggested that the FXII:C and FXII:Ag of p.Glu502Lys variant in the supernatant were 28% and 24%, compared with the wild-type (100%) and FXII:Ag of cell lysates was 39% compared to the wild-type (100%). The FXII:C and FXII:Ag of p. Gly542Ser variant in the supernatant were 32% and 17% and the FXII:Ag of cell lysates was 59%. CONCLUSION The 46TT genetype, p.Glu502Lys and p.Gly542Ser variants of the F12 gene probably underlie the low FXII level in the proband. As shown by in vitro experiment, the p.Glu502Lys and p.Gly542Ser variants can both inhibit the synthesis and secrection of the FXII protein.
Exons ; Factor XII ; genetics ; Factor XII Deficiency ; genetics ; Heterozygote ; Humans ; Pedigree

Objective: Subthreshold attention-deficit/hyperactivity disorder (ADHD) has been suggested to be a “morbid condition” which also needs medical attention. Methods: The present study recruited 89 children with subthreshold ADHD (sADHD), 115 children with diagnosed ADHD (cADHD), and 79 healthy controls (HC) to explore the clinical manifestation, executive functions (EFs) of sADHD, and the caregiver strain. The clinical manifestation was evaluated through clinical interviews and parent-reports. Executive functions were assessed both experimentally and ecologically. Caregiver strain was measured by a parent-reported questionnaire. Results: For the clinical manifestation, both sADHD and cADHD indicated impairments when compared with HC. The comorbidities and the scaled symptoms indicated that the externalizing behaviors were relatively less serious in sADHD than cADHD, whereas the internalizing behaviors between two groups were comparable. For ecological EFs, sADHD scored between cADHD and HC in inhibition and working memory. For experimental EFs, sADHD was comparable to cADHD in inhibition, shifting, and was worse than cADHD in verbal working memory. For the caregiver strain, all scores of sADHD were between that in cADHD and that in HC. Conclusion Our present findings supported the suggestion of subthreshold ADHD as “morbid condition,” which should be treated with caution in clinical practice, especially for the internalizing behaviors and some key components of EFs.

Objective To investigate the potential of polylactic acid glycolic acid copolymer (PLGA) composite ordered multi tunnel collagen scaffold in fabricating a biomimetic artificial nerve graft to repair the sciatic nerve defects in rats.Methods The ordered multi tunnel collagen scaffold was prepared by vacuum freeze-drying and directional drawing method to simulate the epineurium;the outer conduit was prepared by PLGA to simulate the epineurium;and then,the ordered multi tunnel collagen scaffolds were loaded in the PLGA conduit (5∶1) under a stereomicroscope to develop a novel biomimetic artificial nerve.Sixty-four rats were randomly divided into four groups:artificial nerve group,PLGA group,peripheral nerve group,and non-graft control group (n=16);rats in the artificial nerve group,PLGA group,and peripheral nerve group were repaired with artificial nerve graft,hollow PLGA conduit and allogeneic sciatic nerve to bridge the sciatic nerve defect,while the sciatic nerve with the gap in rats of the control group was without any grafting.After 11 weeks of operation,the hind limbs of rats in each group were detected by behavioral test,electrophysiological examination and Fluoro-Gold retrograde tracing method.The changes of muscle tissues (gastrocnemius) were observed by hematoxylin staining and TMR-α-BTX staining,and the regenerated axons were observed by immunohistochemical staining with NF200 and the regenerative spinal anterior horn motor neurons were observed by Nissl fluorescence staining 12 weeks after operation.Results After 11 weeks of operation,the recoveries of the motor functions (the distance between the injured hindlimb and forelimb,the rotation angle of the injured foot) in the peripheral nerve group,artificial nerve group,PLGA group and control group were significantly deteriorated in turn,and the differences were statistically significant (P＜0.05).Electrophysiological examination showed that the recovery effect of peripheral nerve group was the best in both latency and amplitude of the compound muscle action potential,followed by artificial nerve group.The latency of PLGA group was the longest and the amplitude of compound action potential was the smallest;significant differences were noted between each two groups (P2＜0.05).At 12 weeks after operation,the wet weight ratio of muscle fibers,area of muscle fibers and neuromuscular junction area were significantly different between each two groups (P＜0.05);the degree of gastrocnemius atrophy in the artificial nerve group was significantly improved than that in the PLGA group,but not yet reached the level of peripheral nerve group.NF200 immunohistochemical staining showed that a large number of NF200-positive axons were seen in the grafts of the artificial nerve group,but the number was slightly smaller than that of the peripheral nerve group;the number of regenerated axons in the PLGA group was the smaller and mainly distributed near the proximal side.In the PLGA group,only (19.33 ±6.73)％ regenerated spinal anterior horn motor neurons were labeled with Fluoro-Gold,while the positive rates of Fluoro-Gold in the artificial nerve group and peripheral nerve group were (42.67±7.45)％ and (50.13±4.33)％;the differences between each two groups were statistically significant (P＜0.05).Conclusion The biomimetic artificial nerve made of PLGA conduit and ordered multi tunnel collagen scaffold can efficiently reconstruct the defected peripheral nerve with guiding axonal regeneration and promoting functional restoration in rats;however,its effect is poor than peripheral nerve grafting.

Gene editing technology has been a hotspot in the field of biotechnology. CRISPR/Cas systems are efficient gene editing tools because of its specificity, simplicity and flexibility, these features enabled the rapid application of CRISPR/Cas systems in a variety of organisms. Moreover, the combination of transcriptional activator with dead Cas protein can achieve specific regulation of gene expression at the transcription level, which has made important contributions to the development of biotechnology in medical and agriculture. Overexpression of foreign genes is a common method to verify gene function and regulation. However, due to the limitation of vector capacity, it is difficult to achieve overexpression of multiple genes. CRISPR/Cas9 activation system can regulate the expression of multiple genes under the guidance of different guide RNAs to verify gene functions at the regulatory level. This review summarizes the composition of the CRISPR/Cas9 activation system and different activation strategies, and summarizes solutions for excessive activation. It may facilitate the application of CRISPR/Cas9 activation system in genetic improvement of cotton and herbicide resistance research.
Biotechnology ; CRISPR-Cas Systems/genetics* ; Gene Editing ; Phenotype ; RNA, Guide, Kinetoplastida/metabolism*

Specific pathogen-free (SPF) chickens are widely used in the research of avian diseases and vaccines. Vertically transmissible diseases are transmitted to chickens through vertical transmission, seriously affecting their survival rate, increasing production costs, and causing significant economic losses to the poultry industry, while severely impacting the breeding and use of SPF chickens. Therefore, it is crucial for researchers and managers to enhance their understanding of vertically transmissible pathogens in chickens and to develop effective monitoring measures. Quality monitoring is an important part of ensuring the quality of SPF chickens, with pathogen detection being the primary step. Based on this, it is necessary to cultivate qualified SPF chickens through purification methods and biosecurity measures. This paper reviews the major vertically transmissible pathogens in chickens, including viral pathogens, bacterial pathogens and mycoplasmas, as well as their detection methods. This study compares the differences in microbiological testing items and methods for SPF chickens between the U.S. corporate standard and the Chinese national standard. Analysis of the results shows that in both standards, vertically transmissible pathogens such as Escherichia coli, Proteus mirabilis, Salmonella, and avian leukosis are not included in the microbiological testing items for SPF chickens. Instead, these pathogens are characterized by mixed infections, and outbreaks can seriously affect flock health. To produce higher-quality SPF chickens, it is necessary to include these pathogens in the mandatory testing items. The aim of this paper is to help readers understand the relevant standards for microbiological monitoring of SPF chickens, the hazards of vertically transmissible pathogens, and prevention and control strategies, so as to provide a reference for the detection and purification of pathogens in SPF chickens.