Cellulose synthase (CesA), one of the key enzymes in the biosynthesis of cellulose in plants, plays an important role in plant growth and plant resistance. In this study, a total of 21 AsCesA genes from Aquilaria sinensis were systematically identified and the physico-chemical characteristics were analyzed based on genome database and bioinformatical methods. The phylogenetic tree was constructed and the gene location on chromosome, cis-acting elements in the 2 000 basepairs upstream regulatory regions and conservative motifs were analyzed. The AsCesA proteins were mainly located on the plasma membrane. The number of amino acids of the proteins ranged from 390 to 1 261. The isoelectric point distributed from 5.67 to 8.86. All of the 21 AsCesA proteins possessed the transmembrane domains, the number of which was from 6 to 8. The genes were classified into 3 groups according to the phylogenetic relationship. Obvious differences were observed in motif composition in genes from different groups. However, motif2, motif6, motif7 and motif10 were observed in all of AsCesA proteins. Analysis of cis-acting elements indicated that AsCesA genes family has cis-acting elements related to plant hormones, abiotic stresses, and biological processes. Seven AsCesA genes with differential expression were selected according to the calli transcriptome data induced by NaCl at different times and their expression levels under different abiotic stresses were analyzed by quantitative real-time PCR. The results indicated that salt, low temperature, drought, and heavy metal stresses could affect the expression level of AsCesA genes, and the abundance of AsCesA1, AsCesA3 and AsCesA20 showed a significant change, implying their potential important roles to the abiotic stresses. The accumulation pattern of cellulose content under different abiotic stresses was similar to the expression trend of AsCesA genes. Our results provide valuable insights into the role of cellulose synthase in A.sinensis in plant defense.

Objective To investigate the relationship between the degree of liver fibrosis in non-alcoholic fatty liver disease(NAFLD)and the left ventricular structure and function of the heart.Methods A total of 411 patients with NAFLD admitted to the Sec-ond Affiliated Hospital of Xinjiang Medical University from June 2022 to April 2023 were collected.NAFLD patients were stratified using the FIB-4 non-invasive liver fibrosis scoring system and divided into the progressive fibrosis group(n=138)and non-progressive fi-brosis group(n=273).A total of 151 patients with normal abdominal ultrasound examination during the same period were selected as the control group,the relationship between the severity of liver fibrosis in NAFLD patients and the left ventricular structure and function of the heart were analyzed.Results Liver fibrosis was a risk factor for cardiac diastolic function limitation in NAFLD patients(P＜0.05),and the more severe the degree of liver fibrosis,the more severe the cardiac diastolic function limitation.Conclusion The degree of liver fi-brosis in NAFLD is significantly correlated with limited left ventricular diastolic function;it may be related to changes in the left ventricu-lar structure of the heart.

Glyceryl phenylbutyrate(GPB)serves as a long-term management medication for Ornithine transcarbamylase deficiency(OTCD),effectively controlling hyperammonemia,but there is a lack of experience in using this medicine in China.This article retrospectively analyzes the case of a child diagnosed with OTCD at Shanghai Children's Medical Center,Shanghai Jiao Tong University School of Medicine,including a review of related literature.After diagnosis,the patient was treated with GPB,followed by efficacy follow-up and pharmacological monitoring.The 6-year and 6-month-old male patient exhibited poor speech development,disobedience,temper tantrums,and aggressive behavior.Blood ammonia levels peaked at 327 μmol/L;urine organic acid analysis indicated elevated uracil levels;cranial MRI showed extensive abnormal signals in both cerebral hemispheres.Genetic testing revealed de novo mutation in the OTC gene(c.241T＞C,p.S81P).Blood ammonia levels were approximately 43,80,and 56 μmol/L at 1,2,and 3 months after starting GPB treatment,respectively.During treatment,blood ammonia was well-controlled without drug-related adverse effects.The patient showed improvement in developmental delays,obedience,temperament,and absence of aggressive behavior.

Dihydroflavonol 4-reductase (DFR) plays an essential role in the biosynthesis of anthocyanin and regulation of plant flower color. Based on the transcriptome data of Cistanche tubulosa (Schenk) Wight, a full-length cDNA sequence of CtDFR gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR). CtDFR contains an open reading frame (ORF) of 1 263 bp which encodes 420 amino acids with a predicted molecular weight of 47.5 kDa. The sequence analysis showed that CtDFR contains a nicotinamide adenine dinucleotide phosphate (NADPH) binding domain and a specific substrate binding domain. The expression analysis indicated that CtDFR was highly expressed in red and purple flowers, and the relative expression levels were 4.04 and 19.37 times higher than those of white flowers, respectively. The recombinant CtDFR protein was expressed in E.coli BL21 (DE3) using vector pET-28a-CtDFR and was purified. In vitro enzyme activity analysis, CtDFR could reduce three types of dihydroflavonols including dihydrokaempferol, dihydroquercetin, and dihydromyricetin to leucopelargonidin, leucocyanidin and leucodelphinidin. Subcellular localization analysis showed that CtDFR was mainly localized in the cytoplasm. These results demonstrate that CtDFR plays an important role in regulation of flower color in C. tubulosa and make a valuable contribution for the further investigation on the regulation mechanism of C. tubulosa (Schenk) Wight flower color.

Objective: To explore the characteristics of plasma Epstein-Barr virus (EBV) DNA in primary infection in pediatric cases. Methods: The laboratory and clinical data of 571 children diagnosed with EBV primary infection in Children's Hospital of Fudan University during September 1^st, 2017 to September 30^th, 2018 were retrospectively analyzed. According to the results of plasma EBV DNA, they were divided into positive group and negative group. According to the EBV DNA, they were devided into high plasma virol load group and low plasma virol load group. The Chi-square test, Wilcoxon rank sum test were used to compare the differences between groups. Results: Among the 571 children with EBV primary infection, 334 were males and 237 were females. The age of first diagnosis was 3.8 (2.2, 5.7) years. There were 255 cases in positive group and 316 cases in negative group. The percentage of cases with fever,hepatomegaly and (or) splenomegaly, elevated transaminase in the positive group were higher than those in the negative group (235 cases (92.2%) vs. 255 cases (80.7%), χ²=15.22, P<0.001; 169 cases (66.3%) vs. 85 cases (26.9%), χ²=96.80, P<0.001; and 144 cases (56.5%) vs. 120 cases (38.0%), χ²=18.27, P<0.001; respectively).In the positive group, 70 cases were followed up for 46 (27, 106) days, 68 cases (97.1%) turned negative within 28 days, with the exception of 2 cases (2.9%) developed chronic active EBV infection by follow-up revision.There were 218 cases in high plasma viral DNA copies group and 37 cases in low copies group. More cases presented with elevated transaminases in the high plasma viral DNA copies group than those in the low group (75.7% (28/37) vs. 56.0%(116/207), χ²=5.00, P=0.025).Both the positive rate of EBV DNA in peripheral blood leukocytes (84.2% (266/316) vs. 44.7% (255/571), χ²=76.26, P<0.001) and the copies of EBV DNA (7.0×10⁷ (1.3×10⁷, 3.0×10⁸) vs. 3.1×10⁶ (1.6×10⁶, 6.1×10⁶) copies /L, Z=15.23, P<0.001) were higher than that of plasma. Conclusions: In immunocompetent pediatric cases diagnosed as EBV primary infection, cases with positive plasma EBV DNA were prone to have fever, hepatomegaly and (or) splenomegaly, and elevated transaminase than those with negative plasma viral DNA. The plasma EBV DNA usually turns negative within 28 days after initial diagnosis.Most cases with high viral load in plasma showed elevated aminotransferase.
Female ; Male ; Humans ; Child ; DNA, Viral ; Herpesvirus 4, Human ; Epstein-Barr Virus Infections ; Hepatomegaly ; Retrospective Studies ; Splenomegaly ; Fever ; Transaminases

Tbx18,Wt1,and Tcf21 have been identified as epicardial markers during the early embryonic stage.However,the gene markers of mature epicardial cells remain unclear.Single-cell transcriptomic analysis was performed with the Seurat,Monocle,and CellphoneDB packages in R software with standard pro-cedures.Spatial transcriptomics was performed on chilled Visium Tissue Optimization Slides(10x Genomics)and Visium Spatial Gene Expression Slides(10x Genomics).Spatial transcriptomics analysis was performed with Space Ranger software and R software.Immunofluorescence,whole-mount RNA in situ hybridization and X-gal staining were performed to validate the analysis results.Spatial transcriptomics analysis revealed distinct transcriptional profiles and functions between epicardial tissue and non-epicardial tissue.Several gene markers specific to postnatal epicardial tissue were identified,including Msln,C3,Efemp1,and Upk3b.Single-cell transcriptomic analysis revealed that cardiac cells from wildtype mouse hearts(from embryonic day 9.5 to postnatal day 9)could be categorized into six major cell types,which included epicardial cells.Throughout epicardial development,Wt1,Tbx18,and Upk3b were consistently expressed,whereas genes including Msln,C3,and Efemp1 exhibited increased expression during the mature stages of development.Pseudotime analysis further revealed two epicardial cell fates during maturation.Moreover,Upk3b,Msln,Efemp1,and C3 positive epicardial cells were enriched in extracellular matrix signaling.Our results suggested Upk3b,Efemp1,Msln,C3,and other genes were mature epicardium markers.Extracellular matrix signaling was found to play a critical role in the mature epicardium,thus suggesting potential therapeutic targets for heart regeneration in future clinical practice.

OBJECTIVE: To investigate the role of tacrolimus-binding protein 38 (FKBP38) in follicle development and the mechanism by which Fkbp38 gene deletion causes premature ovarian insufficiency (POI). METHODS: The Cre-loxp system was used to construct oocyte-specific Fkbp38 knockout transgenic mice. The genotype of the transgenic mice was identified using PCR, and the expression of FKBP38 in the oocytes was verified. The numbers of primordial follicles, primary follicles, secondary follicles and antral follicles in Fkbp38 knockout mice and non-transgenic littermate control mice were counted with HE staining under a microscope for analyzing the effect of Fkbp38 deletion on follicular development. The fertility and serum sex hormone levels of the mice were determined by reproduction experiments and ELISA to assess ovarian function. Ovarian granulosa cell apoptosis of the mice was assessed using TUNEL assay. The activity of the downstream target protein of phosphorylated ribosomal S6 (PS6) of mTOR signaling pathway was detected, and the expressions of BCL-2 and BAX proteins were determined using immunofluorescence assay for assessing oocyte development in the mice. RESULTS: The oocyte-specific Fkbp38 knockout transgenic mouse model was successfully constructed, which showed decreased fertility, disordered sex hormone levels, and significantly reduced primordial follicles, primary follicles and secondary follicles in the ovary (P < 0.05), demonstrating POI-like changes. Compared with the control mice, oocyte-specific Fkbp38 knockout caused activation of the mTOR signaling pathway, significantly increased apoptosis of the granulosa cells, and obviously increased the BAX/BCL- 2 ratio by increasing BAX expression and reducing BCL-2 expression in the oocytes (P < 0.05). CONCLUSION FKBP38 plays an important role in follicle development, and Fkbp38 gene deletion in mice causes POI possibly by activating the mTOR signaling pathway and inducing granulosa cell apoptosis.
Female ; Humans ; Mice ; Animals ; Primary Ovarian Insufficiency/genetics* ; Apoptosis ; Signal Transduction ; Proto-Oncogene Proteins c-bcl-2 ; Granulosa Cells ; Mice, Transgenic ; Mice, Knockout ; TOR Serine-Threonine Kinases

Objective:To explore the effect of mini-clinical evaluation exercise (MiniCEX) and direct observation of procedural skills (DOPS) formative evaluation in standardized residency training of respiratory and critical care medicine.Methods:The residents who participated in standardized residency training of Zhongshan Hospital, Fudan Unviersity in 2019 were collected in study group, and the residents received the training in previous were collected in control group. The graduation examination scores, the effect of progressive rotations and the satisfaction of residents and examiners were compared between the two groups.Results:The scores of medical history writing, operation and clinical abilities were higher in the study group who were received MiniCEX-DOPS formative evaluation than those in the control group ( P<0.05). The scores of MiniCEX-DOPS evaluation in the third rotation were better than those in the first rotation ( P<0.05). The residents and examiners were generally satisfied with MiniCEX-DOPS evaluation. Conclusion:MiniCEX-DOPS evaluation is effective in residents training of respiratory and critical care medicine, and it is worth popularizing.

Animals ; Hindlimb ; Male ; Mice ; Mice, Inbred C57BL ; Muscle Fibers, Skeletal ; Muscle, Skeletal/pathology* ; Muscular Atrophy

Objective:To analyze the correlation between obstructive sleep apnea (OSA) and attention deficit hyperactivity disorder (ADHD).Methods:The clinical Data, polysomnography (PSG) and cognitive function examination results of 112 OSA children admitted to Department of Otorhinolaryngology Head and Neck Surgery of the Second Affiliated Hospital of Xi′an Jiaotong University from January 2019 to June 2020 were retrospectively analyzed. According to the severity of OSA, the children were divided into mild, moderate and severe OSA groups, and the basic demographic characteristics, sleep parameters and ADHD occurrence were analyzed. According to the results of ADHD examination, the children were divided into ADHD group and non-ADHD group, and the basic demographic characteristics and sleep parameters were analyzed. Taking these parameters as independent variables, binary Logistic regression analysis was conducted to establish the model equation for predicting the risk of OSA associated ADHD among children.Results:Grouped by OSA severity, among the three groups, apnea-hypopnea index (AHI) [3.70 (2.84, 5.47) vs 8.59 (7.50, 9.54) vs 19.48 (15.83, 25.23)], obstructive apnea index (OAI) [1.31 (0.93, 1.82) vs 3.03 (1.54, 4.41) vs 11.69 (8.53, 15.42)], obstructive apnea-hypopnea index (OAHI) [2.82 (1.81, 3.64) vs 6.17 (5.58, 7.26) vs 15.68 (13.12, 21.25)], and respiratory event-related arousal index [0.50 (0.25, 1.05) vs 1.25 (0.70, 2.23) vs 2.40 (1.60, 4.70)] increased, minimum pulse oxygen saturation (SpO 2) [90.00 (88.00, 92.00) vs 87.00 (83.00, 90.25) vs 81.00 (76.00, 85.00)] decreased, the differences were statistically significant (all P<0.05). The non-rapid eye movement (NREM)1 period time ratio of the severe OSA group was significantly longer than that of the mild OSA group, while the average SpO 2 was significantly lower than that of the mild OSA group; the NREM3 period time ratio of the moderate and severe OSA group was significantly less than that of the mild OSA group; the arousal index of the severe OSA group was significantly greater than the mild or moderate OSA group. There were no statistically significant differences among the three groups in gender, age, body mass index, sleep efficiency, rapid eye movement (REM) period time ratio, and NREM2 period time ratio (all P>0.05). Mild OSA group had 10 cases of ADHD (17.54%), moderate OSA group had 7 cases (23.33%) of ADHD, severe OSA group had 9 cases of ADHD (36.00%), and the difference was not statistically significant. Grouped by ADHD examination, the AHI, OAI, OAHI, and NREM1 period time ratios of the ADHD group were significantly higher than those of the non-ADHD group, while the sleep efficiency, minimum SpO 2 and NREM3 period time ratio were significantly lower than those of the non-ADHD group. The Logistic regression analysis suggested that ADHD was correlated with sleep efficiency, minimum SpO 2, and NREM3 period time.The established Logistic regression equation was: X=15.670+0.061×(sleep efficiency)-0.212×(minimum SpO 2)-0.144×(NREM3 period time ratio), the sensitivity and specificity of the model prediction were 84.6% and 79.1% respectively when the area under the receiveroperating characteristic curves was 0.867. Conclusions:OSA and ADHD in children have a certain correlation. Sleep structure disturbance and intermittent hypoxia may be important reasons. The predictive model equations obtained by PSG in this study can be used to assess the risk of ADHD in children with OSA.