OBJECTIVE: To explore the clinical phenotype and genetic variant in a child with Verheij syndrome (VRJS). METHODS: A child who had presented at the Soochow University Affiliated Children's Hospital and Wujiang District Children's Hospital in July 2022 for "elevated scapula since early childhood" was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing. Candidate variant was verified by Sanger sequencing and bioinformatic analysis. RESULTS: The child had manifested elevated scapulae, torticollis, neck asymmetry, facial dysmorphism, dispersed café-au-lait spots, limited mobility of upper limbs and shoulder joints, and intellectual disability. Sequencing revealed that he has harbored a de novo heterozygous c.405dupT (p.Ile136Tyrfs*4) variant of the PUF60 gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), this variant was classified as pathogenic (PVS1+PS2_moderate+PM2_supporting). Combined his clinical features and result of genetic testing, the child was diagnosed with VRJS due to variant of the PUF60 gene. CONCLUSION The clinical manifestations of VRJS include facial dysmorphism, intellectual disability, elevated scapulae, vertebral fusion, other skeletal malformations, without significant abnormalities of the heart, kidney, and eyes, which need to be distinguished from Klippel-Feil syndrome. Above finding has expended the mutation spectrum of the PUF60 gene and provided a reference for delineation of the genotype-phenotype correlation of the VRJS.
Child ; Child, Preschool ; Humans ; Male ; Cafe-au-Lait Spots ; Computational Biology ; Genetic Testing ; Genomics ; Intellectual Disability/genetics* ; Mutation

Objective:To explore the relevant predictive indicators of fever course > 7 days in children with infectious mononucleosis.Methods:The clinical data of 163 children with infectious mononucleosis who received treatment in Xi'an Children's Hospital from January 2018 to October 2020 were retrospectively analyzed. According to the heat duration, the children were divided into the fever course > 7 days group ( n = 55) and the fever course ≤ 7 days group ( n = 108). The clinical manifestations and laboratory indexes on admission were compared between the two groups. A logistic regression model was used to analyze the influential factors of fever course in children. A receiver operating curve was used to evaluate the predictive value of heat course > 7 days for infectious mononucleosis. Results:The majority of children with infectious mononucleosis had a heat course of 7 days (21.5%). There were no significant differences in clinical manifestations between the fever course > 7 days group and the fever course ≤ 7 days group (all P > 0.05). Neutrophil count, the proportion of monocytes, aspartate aminotransferase, and the proportion of suppressor T (Ts) cells in the fever course > 7 days group were (15.97 ± 7.60) × 10 9/L, 7.75 (4.93, 10.75)%, 53.00 (22.00, 91.50) U/L, 70.00 (57.00, 75.00)%, respectively, which were significantly higher than (15.21 ± 5.29) × 10 9/L, 5.40 (3.40, 9.60)%, 40.00 (30.00, 63.75) U/L, 63.50 (55.00,70.75)% in the fever course ≤ 7 days group ( t = -5.10, Z = -2.31, Z = -2.26, Z = -2.12, all P < 0.05). The proportion of helper T (Th) cells and the ratio of Th/Ts cells in the fever course > 7 days group were 13.00 (9.00, 17.00)% and 0.19 (0.12, 0.30)%, respectively, which were significantly lower than 16.00 (12.25, 20.75)%, 0.26 (0.18, 0.37)% in the fever course ≤ 7 days group ( Z = 2.44, 2.48, both P < 0.05). Multivariate logistic regression analysis showed that the increased proportion of Ts cells ( OR = 0.96, 95% CI 0.922-0.978, P < 0.05) was an influential factor of the prolonged course of fever. The area under the receiver operating characteristic curve of the proportion of Ts cells was 0.637. The cut-off value, sensitivity, and specificity were 67.50%, 61.3%, and 64.3%, respectively. Conclusion:Children with infectious mononucleosis with a longer heat course have more severe immune responses. The proportion of Ts cells > 67.5% can be used as a risk factor for the fever course > 7 days in children with infectious mononucleosis.

Objective:To explore the mediating effects of mindfulness and perceived stress between childhood trauma and psychotic-like experiences.Methods:In October 2021, 602 freshmen from a university in Guangdong Province were tested by childhood trauma questionnaire(CTQ), mindful attention awareness scale(MAAS), perceived stress scale(PSS), and community assessment of psychic experiences(CAPE). SPSS 21.0 and Mplus 8.3 were used for data analysis. Mann-Whitney U test was used to compare the differences between groups, and Spearman correlation analysis was used to investigate the correlation among childhood trauma, mindfulness, perceived stress and psychotic-like experiences. Bias-corrected nonparametric percentile Bootstrap method was used to examine the mediating roles of mindfulness and perceived stress between childhood trauma and psychotic-like experiences. Results:(1) In the past one month, 59.5% of the freshmen had psychotic-like experiences, and 8.5% of them experienced the distress. (2)Childhood trauma was positively correlated with perceived stress( r=0.29, P<0.01), frequency ( r=0.14, P<0.01)and distress( r=0.17, P<0.01) of psychotic-like experiences, and was negatively correlated with mindfulness( r=-0.28, P<0.01). (2)Mindfulness and perceived stress played individual mediating effects and chain mediating effect between childhood trauma and frequency of psychotic-like experiences, with effect size of 0.08, 0.04, 0.03, accounting for 53.33%, 26.67%, 20.00% of the total indirect effect(effect size=0.15), respectively. Mindfulness and perceived stress played individual mediating effects and chain mediating effect between childhood trauma and distress of psychotic-like experiences, with effect size of 0.07, 0.04, 0.03, accounting for 50.00%, 28.57%, 21.43% of the total indirect effect(effect size=0.14), respectively. Conclusion:Childhood trauma can affect psychotic-like experiences and distress through the indirect effects of mindfulness and perceived stress, or through the chain mediating effect of mindfulness-perceived stress.

Objective:To investigate the application of artificial intelligence based on the neural network radiation field in repair of soft tissue defects at lower limbs.Methods:A retrospective analysis was performed of the 23 patients who had been admitted to Department of Orthopedic Surgery, Renmin Hospital of Wuhan University from June 2020 to May 2022 for soft tissue defects at lower limbs. There were 14 males and 9 females, aged (38.6±6.7) years. Causes for soft tissue defects: traffic injury in 9 cases, benign or malignant primary soft tissue tumor in 6 cases, mechanical injury in 4 cases, crush injury in 2 cases, and chronic ulcer in 2 cases. Defect locations: the thigh in 3 cases, the lower leg in 7 cases, and the ankle and distal foot in 13 cases. The areas of soft tissue defect ranged from 6.0 cm×3.8 cm to 14.7 cm×12.8 cm. The defects were repaired and reconstructed by transplantation of an anterolateral femoral free flap in 7 cases and a pedicled flap in 16 cases with the assistance of artificial intelligence based on the neural network radiation field, a cutting-edge artificial intelligence algorithm that can quickly construct and process three-dimensional model images through volume rendering under the radiation field. The flap survival rate, aesthetic satisfaction before and after treatment, time for skin flap harvesting and transplantation, functional recovery of lower limbs and incidence of complications were recorded.Results:All the 23 patients were followed up for 32(28, 36) weeks. All the flaps were harvested smoothly and survived. The time for flap harvesting and transplantation was 65.8(50.0, 76.0) min. The aesthetic satisfaction scored (2.3±0.7) points before treatment and (8.4±1.6) points 4 weeks after treatment, showing a statistically significant difference ( P<0.05). The skin flaps healed well with no complications such as hematoma or infection in all but one patient who suffered from superficial necrosis at the distal skin flap due to venous crisis but healed with a scar. On average, the functional recovery of lower limbs scored 23.7(22.0, 25.0) points at 12 weeks after operation according to the Enneking evaluation system, and the functional recovery of lower limbs was 79% (23.7/30.0). Conclusion:Application of artificial intelligence based on the neural network radiation field can achieve ideal results in repair of soft tissue defects at lower limbs, due to its advantages of rapid and accurate surgical procedures, limited damage to the donor site, and a short learning curve.

Transketolase (EC 2.2.1.1, TK) is a thiamine diphosphate-dependent enzyme that catalyzes the transfer of a two-carbon hydroxyacetyl unit with reversible C-C bond cleavage and formation. It is widely used in the production of chemicals, drug precursors, and asymmetric synthesis by cascade enzyme catalysis. In this paper, the activity of transketolase TKTA from Escherichia coli K12 on non-phosphorylated substrates was enhanced through site-directed saturation mutation and combined mutation. On this basis, the synthesis of tartaric semialdehyde was explored. The results showed that the optimal reaction temperature and pH of TKTA_M (R358I/H461S/R520Q) were 32 ℃ and 7.0, respectively. The specific activity on d-glyceraldehyde was (6.57±0.14) U/mg, which was 9.25 times higher than that of the wild type ((0.71±0.02) U/mg). Based on the characterization of TKTA_M, tartaric acid semialdehyde was synthesized with 50 mmol/L 5-keto-d-gluconate and 50 mmol/L non-phosphorylated ethanolaldehyde. The final yield of tartaric acid semialdehyde was 3.71 g with a molar conversion rate of 55.34%. Hence, the results may facilitate the preparation of l-(+)-tartaric acid from biomass, and provide an example for transketolase-catalyzed non-phosphorylated substrates.
Escherichia coli/genetics* ; Transketolase/chemistry* ; Tartrates ; Escherichia coli Proteins/genetics*

Background:Based on the effect of seasonal changes on human visceral function,this study investigated the impact of seasonal photoperiod of the pineal body on hypothalamic-pituitary-adrenal axis-hippo-campal-receptor in rats,aiming to reveal the mechanism by which pineal gland melatonin regulates the seasonal secretion of hippocampal neurotransmitters.Methods:Vernal equinox,summer solstice,autumn equinox,and winter solstice were selected as four experimental time points,and rats were randomly divided into normal control group,sham operation group,and pinealectomized group.The seasonal changes in corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),corticosterone,hypothalamic melatonin receptor(MTR),and hippocampal corticosterone receptor(CORTR)were examined by enzyme-linked immunosorbent assay.Results:Comparing the same group between different seasons,we showed that in the normal control group,CRH,ACTH,corticosterone,and MTR were higher,while CORTR was lower in autumn and winter than in spring(all P＜.05).Compared with the normal control group,the pinealectomized group showed higher levels of corticosterone(P=.01),MTR(P=.01),and CORTR(P=.03)during spring;reduced levels of MTR and CORTR(both P＜.001)during summer;higher levels of ACTH(P=.001)and MTR(P＜.001),and lower levels of CRH(P=.001),corticosterone(P＜.001),and CORTR(P=.003)during autumn;and lower levels of CRH(P＜.001)and MTR(P=.004),and higher level of ACTH(P＜.001)in winter.Conclusions:Seasonal photoperiod acts on the pineal gland to secrete different levels of melatonin,resulting in seasonal changes in the hypothalamic-pituitary-adrenal axis-hippocampal-receptor,which may be the pathophysiological basis for the onset of seasonal affective disorder.

Background: Based on the theory of"five Zang-organs corresponding to the seasons"in traditional Chinese medicine (TCM), physiological functions including emotions vary with the seasons. We aimed to investigate the seasonal photoperiod effects of melatonin (MT) released from the pineal gland on the MT receptor (MTR)-Gs/Gi-cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA)-cAMP response element-binding protein (CREB) signaling pathway in the hippocampus.Methods: Rats were divided into three groups: control, operation (surgery with pineal gland removal), and pseudo-operation groups (same surgery as operation group but without removing pineal gland), and fed at specific time across the four seasons. The levels of MTR, adenylate cyclase (AC), cAMP, PKA, and CREB in the hippocampus were analyzed using ELISA. The concentrations of Gs and Gi were analyzed using Western blot. The expression of CREB mRNA was detected by PCR. Results: For intragroup comparisons, compared with spring, the levels of Gs/Gi in the control group were higher in summer, autumn, and winter (P=.009 in summer;P<.001 in autumn and winter);the levels of MTR, cAMP, PKA, and CREB in the control group were significantly higher in autumn and winter than in spring (all P<.001). The levels of MTR, cAMP, PKA, and CREB in each season were significantly lower in the operation group than in the control group (all P < .05). Significant differences were noted in Gs/Gi levels between the operation group and control group in spring, autumn, and winter (all P<.05). Conclusion: Our findings suggest that MTR-Gs/Gi-cAMP-PKA-CREB signaling pathway is involved in the seasonal photoperiod effects of the pineal gland on the hippocampus and may underpin seasonal changes in emotions. It can support the prevention and treatment of the seasonal onset of mental dis-orders, and enrich the theory of"five Zang-organs corresponding to the seasons".

: Objective: To investigate the effects of ezrin enhancer knockout on ezrin gene expression, cell proliferation and migration of human esophageal carcinoma Eca-109 cells. : Methods: The CRISPR/Cas9 recombinant plasmids targeting upstream/downstream of human ezrin enhancer were co-transfected into human esophageal carcinoma Eca-109 cells, and the cell line Eca-C2 with ezrin enhancer knockout was screened by purinomycin. Then the expression levels of ezrin mRNAand protein in Eca-C2 cells were detected by Real-time quantitative PCR (qPCR) and Western blotting, respectively; The expression levels of MAPK-pathway-related proteins were detected by protein array technology; and the effects of ezrin enhancer knockout on the proliferation and migration of Eca-C2 cells were analyzed by WST-1 method and wound-healing assay, respectively. : Results:The human esophageal carcinoma cell line Eca-C2 with stable ezrin enhancer knockout was established successfully. Compared with control cells, the mRNA and protein expressions of ezrin in Eca-C2 cells were significantly reduced (all P<0.05).Among the 17 detected MAPK pathway related proteins in Eca-C2 cells, 9 proteins (AKT, CREB, GSK3b, MKK6, mTOR, P38, P53, P70S6K and RSK1) were down-regulated, and the cell proliferation and migration were significantly inhibited (all P<0.05). Conclusion: ezrin enhancer knockout can significantly inhibit the cell proliferation and migration of human esophageal carcinoma Eca-109 cells.

Disulfiram (DSF) has been widely used in clinical treatment of alcoholism.To date,in vivo and in vitro experiments have demonstrated that DSF has strong anti-cancer activity and could improve patient's survival,and the underlying mechanism has been elaborated.In addition,it was reported that,during radiotherapy,DSF could protect normal tissue and cells meanwhile enhance the radiosensitivity of tumor cells by forming complexes with copper ions,suppressing cancer stem cells and inhibiting ubiquitin-proteasome system activity in cancer cells.This review summarizes the completed and ongoing clinical trials of disulfiram,and its anti-tumor mechanisms and advances in radiation biology.