Objective To evaluate the application value of whole exome sequencing (WES) in diagnosis of NDDs (neuro-developmental disorders) children.Metheod WES was used for the diagnosis of 35 unexplained NDD children, which admitted to the outpatient and ward of Children′s hospital affiliated to Capital institute of pediatric from November 2015 to November 2016.These children′s clinical data was collected detailedly.Using bioinformatics software tools combining with patient′s phenotype, the candidate genetic/genomic variants of these patients were identified from WES data.The final pathogenicity of genetic/genomic variants was interpreted according to the guideline of the American College of Medical Genetics and Genomics (ACMG), meanwhile, the variants validation and co-separation analysis in the parents and their family members were performed by Sanger sequencing, real time-PCR and multiplex ligation-dependent probe amplification (MLPA).Results 14 pathogenic single nucleotide variants (SNVs) and three pathogenic copy number variations (CNVs) were detected in the 35 NDD children, the detection rate in this study is 48.6%.Among the 14 pathogenic SNVs, 11 of them are the definite NDD-related genes according to OMIM database (such as CHARGE syndrome, Wiedemann-Steiner syndrome, Cockayne syndrome, etc.), and six of them are de novo (6/11, 54.6%).Three pathogenic CNVs were identified from WES data, including two microduplications and one microdeletion.Meanwhile, a female child carrying a frame shift mutation in MECP2 was found and the germline mosaicism with low-frequency mutation of this site (8.4%) was confirmed by his father's sperm.Conclusions The diagnosis rate of WES in NDDs children is 48.6% in our small-sample study.In addition to pathogenic/likely pathogenic SNVs, CNVs can be detected successfully from WES data, which effectively improved the diagnosis yield in NDDs children.

Objective To investigate the expression and significance of malonaldehyde (MDA), superoxide dismutase (SOD) and endotoxin (ET) in liver injury model of Budd-Chiari syndrome (BCS) in rats. Methods The animal model of BCS was established by partially ligating the inferior vena cava of the posterior segment of liver in rats. The experimental animals were divided into three groups: control group (12 rats), model group (48 rats) and sham operation group (48 rats). The model group and sham operation group were divided into four subgroups (1, 3, 6, 12 weeks) of 12 rats each. After the success of modeling,being confirmed by digital subtraction angiography (DSA), nine rats in each group were sacrificed at random respectively, where their serums and liver tissues was collected. The levels of MDA, SOD and ET in both liver homogenate and serum were examined respectively. ANOVA was used to compare the total difference between groups and within group of each measurement data. The LSD method was used to do multiple comparison within group and between groups. Pearson method was used to do correlation analysis of hypoxia markers. Results The levels of MDA, SOD and ET in liver homogenate and serum at different time points in model group were significantly different from those in control group and sham operation group (MDA: liver homogenate (F=52.906, 219.016), serum (F=21.573, 43.878); SOD: liver homogenate (F=22.927, 19.317), serum (F=10.841, 31.643);ET: liver homogenate (F=33.588, 105.515), serum (F=40.832, 46.323);P＜0.05). The total difference of the MDA level in serum at each time point after the operation was not statistically significant in model group(F=1.965,P=0.139), but that of liver homogenate in the model group was statistically significant (F=7.716, P=0.001). The SOD and ET levels in both liver homogenate and serum of model group were compared within groups at different time points after operation respectively, and the overall difference was statistically significant (SOD: F=17.053, 7.903; ET: F=19.870, 39.372; P＜0.05). The time-varying curves of MDA and ET in liver homogenate and serum in model group were similar, which both increased from 1 week after operation,peaked at 6th week and slightly decreased at 12th week. The increase levels of MDA and ET in liver homogenate were significantly higher than those in serum. There was a negative correlation between MDA and SOD in liver homogenate and serum (r=－0.814,－0.591;P=0.001, 0.001), a positive correlation between MDA and ET (r=0.761, 0.422; P=0.004, 0.001), and a negative correlation between SOD and ET (r=－0.726,－0.490;P=0.001, 0.001). Conclusions The levels of hypoxia related markers, such as MDA, SOD and ET in liver and serum of BCS animal model, change to varying degrees in the early stage, and will be aggravated as the disease continues to advance. In the later stage, with the establishment of collateral circulation, hypoxia will be slightly eased, but is still significantly higher than normal, which indicates that congestion and hypoxia run through the whole process of BCS, and could be the key and initiating factors.

Objective To investigate the influencing factor of the morphology of unruptured intracranial aneurysms for aneurysm wall enhancement under the high-resolution magnetic resonance imaging. Methods From January 2015 to December 2016,the clinical and imaging data of 68 consecutive patients with unruptured intracranial aneurysm (86 aneurysms) in Changhai Hospital,the Second Military Medical University were enrolled retrospectively. Vascular wall imaging technology was used to conduct aneurysm scan,and the aneurysm wall enhancement was identified by the imaging features before and after contrast enhancement. They were divided into either an enhancement group ( n=32,34 aneurysms) or a non-enhancement group (n=45,52 aneurysms) according to whether having the abnormal enhancement of aneurysm wall or not ( because some patients also have enhanced aneurysms and non-enhanced aneurysms, the number of cases of the enhanced or not was calculated seperately in both groups ) . Morphological parameters were calculated by 3D image data,including aneurysm size,ratio of height to width,volume ratio, dome-to-neck ratio, transverse length ratio, bottleneck factor, and inflow angle. Univariate and multivariate logistic analyses were used to determine the morphological influence factors of aneurysm wall enhancement. Results (1) A total of 34 (39. 5%) aneurysms had aneurysm wall enhancement and 52 (60. 5％) aneurysms did not have aneurysm wall enhancement. There were no significant differences in sex, age, hypertension,diabetes, smoking, family history of subarachnoid hemorrhage, and aneurysm site in both groups (all P>0. 05). (2) The aneurysm size,ratio of height to width,volume ratio,dome-to-neck ratio, and bottleneck factor in the enhancement group were larger than those of the non-enhancement group. There were significant differences between the 2 groups (9. 19 [6. 54,11. 04] mm vs. 5. 31 [4. 17,7. 37] mm, (1. 18 [1. 01,1. 69] vs. 0. 91 [0. 72,1. 25],(3. 62 [2. 30,4. 63] vs. 2. 18 [1. 37,2. 76],1. 52 [1. 25, 1. 99] vs. 1. 19 [1. 03,1. 51],and 1. 21 [1. 11,1. 69] vs. 1. 05 [0. 94,1. 31],all P<0. 01). The proportion of irregular morphologic aneurysms in the enhancement group was higher than that in the non-enhancement group. There was significant difference between the 2 groups (55. 9％ [19/34] vs. 17. 3％ [9/52],P<0. 01 ) . There were no significant differences in transverse length ratio and inflow angle between the 2 groups (all P>0. 05). (3) Because the ratio of height to width,volume ratio,dome-to-neck ratio,and bottleneck factor were related to the aneurysm size,the aneurysm size,inflow angle,and irregular shape were included in the multivariate logistic regression analysis. The results showed that aneurysm size ( OR,3. 727,95％CI 1. 933-6. 971,P<0. 01) and irregular shape (OR,3. 990,95％CI 1. 219-13. 065,P=0. 022) were the independent risk factors for aneurysm wall enhancement. Conclusions The size and irregular shape of unruptured intracranial aneurysms are the independent risk factors for aneurysm wall enhancement. High-resolution magnetic resonance wall imaging may become an effective and noninvasive imaging method for evaluating the ruptured risk of intracranial aneurysms.

Objective:To construct three image recognition models of manikin′s glottis using visual laryngoscopy based on deep-learning algorithm.Methods:The tracheal intubation manikin′s epiglottis was visualized using a videolaryngoscope, and then epiglottis was elevated to expose the glottis and acquire glottic images. A total of 149 images were obtained from various angles and orientations and randomly divided into training set and test set, and the annotation of image data was completed. Three glottal image recognition models of CenterNet, YOLOv3 and YOLOv4 were developed. The training set was used to complete the training of the models, and finally the test set was used to evaluate the model performance.Results:CenterNet, YOLOv3 and YOLOv4 three models were successfully constructed, the mean average precision of CenterNet, YOLOv3 and YOLOv4 was 92.33%, 89.52% and 89.02% respectively, the recall rates were 87.50%, 90.00% and 90.00% respectively, the precision rates reached 97.22%, 94.74% and 94.74% respectively, and the accuracy rates were 90.91%, 85.11% and 88.89% respectively. All three algorithms demonstrated an identical F1 score of 91.00%.Conclusions:The CenterNet, YOLOv3 and YOLOv4 models are successfully constructed, and three recognition models can accurately identify the glottis in the image, with the CenterNet model demonstrating the highest recognition precision.

The medical records of 11 pediatric patients undergoing resection of adrenal cortical carcinoma from January 2012 to January 2019 in our hospital were collected.Anesthesia management for resection of adrenal cortical carcinoma in children was analyzed and investigated.Anesthesia was induced by intravenously injecting atropine 0.01 mg/kg, dexamethasone 2-5 mg, propofol 2-3 mg/kg, sufentanil 0.3-0.5 μg/kg or fentanyl 1-2 μg/kg, rocuronium 0.5 mg/kg or cis-atracurium 0.1-0.2 mg/kg.Radial artery catheterization and femoral vein catheterization were performed under ultrasound guidance.Arterial blood pressure was continuously monitored.The esophageal thermometers probe was placed to continuously monitor body temperature.The catheter was placed to monitor urine volume.Intermittent positive pressure ventilation was performed after endotracheal intubation with the inspiratory oxygen fraction set 60%-100%, oxygen flow rate 2-3 L/min, tidal volume 7-10 ml/kg, ventilation frequency 20-26 times/min, inhalation/respiration ratio 1∶(1.5-2.0) and airway pressure 16-20 cmH 2O, and the end-tidal pressure of carbon dioxide was maintained at 35-45 mmHg.Anesthesia was maintained by inhaling 2%-4% sevoflurane and/or intravenously infusing propofol 0.10-0.15 mg·kg -1·min -1, and continuously infusing remifentanil 0.2-0.5 μg·kg -1·min -1.Hemodynamics was maintained within the normal range, and the bispectral index was maintained at 40-60 during the surgery.Before the tumor was completely removed, 5-10 mg/kg sodium hydrocortisone succinate was intravenously infused.At the end of the operation, sufentanil 0.75-1.00 μg·kg -1·d -1 or fentanyl 7.5-10.0 μg·kg -1·d -1 was continuously infused for postoperative analgesia until 48 h after operation.Operation was smoothly completed with stable anesthesia in all the pediatric patients.The tracheal tube was removed successfully after the operation.All children in this group were discharged from hospital and no death occurred.Anesthesia management for resection of adrenal cortical carcinoma required an appreciation of the clinical characteristics and perioperative pathophysiological changes.Paying attention to the changes in hormone levels during perioperative period and timely adjusting the children′s internal environment to maintain the stability of anesthesia and reduce the stress response were the keys to anesthesia management.

In this study, 10 batches of samples of cultured Bovis Calculus(cow-bezoar) were determined for the investigation of chemical profile of bile acids with the UPLC-QDA method established. The results showed that nine common bile acids, cholic acid(CA), deoxycholic acid(DCA), chenodeoxycholic acid(CDCA), taurocholic acid(TCA), taurodeoxycholic acid(TDCA), taurochenodeoxycholic acid(TCDCA), glycocholic acid(GCA), glycodeoxycholic acid(GDCA) and glycochenodeoxycholic acid(GCDCA), were founded in samples and 7 bile acids were quantified except GDCA and GCDCA. In these samples, unconjugated bile acids, the major type of bile acids, accounted for more than 97% of all types of bile acids. As for unconjugated bile acids, CA was the most major bile acid in cultured cow-bezoar and it was about twice as much as DCA. The relative low-cost method established in the current study is accurate, rapid and sensitive, which is suitable for the studies of other drugs from animal bile.
Animals ; Bile ; Bile Acids and Salts ; Calculi ; Cattle ; Female

A determination method for bilirubin in cultured cow-bezoar was developed in this study, with which the bilirubin in 15 batches of samples was quantified. The samples were first processed with 10% oxalic acid solution for the conversion of bilirubin from conjugated to unconjugated, followed by the extraction with dichloromethane. Then the obtained sample solutions were analyzed at 450 nm by HPLC[chromatographic column: Agilent TC-C_(18)(4.6 mm × 250 mm, 5 μm); mobile phase: acetonitrile and 1% glacial acetic acid aqueous solution(95∶5); flow rate: 1.0 mL·min~(-1)]. The bilirubin content in the 15 batches of cultured cow-bezoar was ranged from 21.9% to 41.7% with the average of 32.4%. The proposed method is accurate and reliable, thus making it suitable for the quantitation of bilirubin in cultured cow-bezoar and its quality assessment and control.
Animals ; Bezoars ; Bilirubin ; Cattle ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Female

Objective:To prepare magnetic solid lipid nanoparticles co-loaded quercetin and resveratrol(QR-MSLN),develop the reasonable characterization method,and investigate its inhibitory effect on transplanted hepatocarcinoma H22 tumor in mice. Method:Magnetic Fe₃O₄ particles coated with oleic acid(OA-Fe₃O₄) were synthesized and its structure was confirmed by Fourier transform infrared spectrometer(FT-IR).QR-MSLN was prepared by emulsion ultrasonic dispersion method,its morphology was examined by transmission electron microscopy,its particle size was determined by laser particle sizer.Concentration of Fe in the preparation was determined by phenanthroline spectrophotometry.The entrapment efficiency,saturation magnetization,in vitro release behavior were investigated by ultrafiltration centrifugation,vibrating sample magnetometer(VSM) and dialysis method,respectively.Mouse tumor model transplanted with hepatoma H22 ascites tumor was established and antitumor effect of QR-MSLN on H22 bearing mice were observed in the presence of an applied magnetic field. Result:Morphology of QR-MSLN was round,and black magnetic particles could be observed inside it,its particle size was (171.9±2.2) nm,the concentration of Fe was (1.40±0.46) g·L^-1.The preparation exhibited apparent superparamagnetism and the saturation magnetization was 7.75 A·m²·kg^-1.The entrapment efficiencies of quercetin and resveratrol in QR-MSLN were 99.10% and 80.83%,respectively.QR-MSLN had a significantly higher effect of tumor inhibition than SLN(containing quercetin and resveratrol) and free drug(P<0.01). Conclusion:QR-MSLN has uniform particle size and good magnetic response,and shows remarkable antitumor effect on H22 bearing mice.

Adult ; Asian Continental Ancestry Group ; Collagen Type II ; genetics ; Exome ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Osteochondrodysplasias ; diagnosis ; genetics ; Pedigree ; Sequence Analysis, DNA

Objective: To explore the association between sleep duration and the risk of frailty among the elderly over 80 years old in China. Methods: Using the data from five surveys of the China Elderly Health Influencing Factors Follow-up Survey (CLHLS) (2005, 2008-2009, 2011-2012, 2014, and 2017-2018), 7 024 elderly people aged 80 years and above were selected as the study subjects. Questionnaires and physical examinations were used to collect information on sleep time, general demographic characteristics, functional status, physical signs, and illness. The frailty state was evaluated based on a frailty index that included 39 variables. The Cox proportional risk regression model was used to analyze the correlation between sleep time and the risk of frailty occurrence. A restricted cubic spline function was used to analyze the dose-response relationship between sleep time and the risk of frailty occurrence. The likelihood ratio test was used to analyze the interaction between age, gender, sleep quality, cognitive impairment, and sleep duration. Results: The age M (Q₁, Q₃) of 7 024 subjects was 87 (82, 92) years old, with a total of 3 435 (48.9%) patients experiencing frailty. The results of restricted cubic spline function analysis showed that there was an approximate U-shaped relationship between sleep time and the risk of frailty. When sleep time was 6.5-8.5 hours, the elderly had the lowest risk of frailty; Multivariate Cox proportional risk regression model analysis showed that compared to 6.5-8.5 hours of sleep, long sleep duration (>8.5 hours) increased the risk of frailty by 13% (HR: 1.13; 95%CI: 1.04-1.22). Conclusion: There is a nonlinear association between sleep time and the risk of frailty in the elderly.
Aged ; Humans ; Aged, 80 and over ; Frailty/epidemiology* ; Sleep Duration ; Prospective Studies ; Sleep/physiology* ; China/epidemiology*