Background and purpose:Liquid biopsy is a kind of blood, urine and other non-solid biologi-cal tissue sampling analysis, mainly for malignant tumor diagnosis, monitoring and predicting its prognosis. In this research, we optimized the extraction of miRNA in urine, established a standardized means of liquid biopsy, screened and verified the miRNA markers in patients with bladder cancer.Methods:From Jan. 2014 to Sept. 2015, we used miRNA microarray in six patients with bladder cancer and six healthy controls. Samples of 78 cases of bladder cancer and 23 healthy controls were tested by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) to verify the relationship between miRNA markers in liquid biopsy and clinical pathological parameters. The diagnostic value of miRNA markers was also analyzed and compared.Results:We screened 10 miRNAs differential expression in urine. Combined with previous literature, we selected 20 miRNAs to verify their expression levels in bladder cancers and healthy controls. miR-509-5p/miR-124 ratio in the urine was found higher in patients with bladder cancer than in healthy controls (P<0.0001). With the rise of miR-509-5p/miR-124 ratio in urine, tumor stage and grade were also increased (P=0.003). When the cutoff was set at 0.41, the diagnostic sensitivity and specificity of miR-509-5p/miR-124 ratio were 73.1% and 82.6%, respectively. The AUC of miR-509-5p/miR-124 ratio to detect bladder cancer was 0.864, higher than that of urinary exfoliated cells (P=0.0002).Conclusion:We optimized the extraction of miRNAs in urine,established a standardized liquid biopsy of miRNA markers. The miR-509-5p/miR-124 ratio could be an ideal diagnos-tic marker for bladder cancer.

OBJECTIVETo develop and evaluate a mouse model of nonalcoholic steatohepatitis (NASH) induced by a high-fat and high-fructose (HFHFr) diet.

METHODSSix-week-old C3H mice were randomly divided into groups for HFHFr diet experimental modeling, high fat-only (HF) diet controls, high fructose-only (HFr) diet controls, and standard chow (SC) diet controls. The standard HFHFr diet was modified so that it consisted of 76.5% standard chow, 12% lard, 1% cholesterol, 5% egg yolk powder, 5% whole milk powder, and 0.5% sodium cholate, along with 20% fructose drinking water. At the end of experimental weeks 4, 8, and 16, measurements were taken for the NASH-related parameters of body mass, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), lipid profile, and wet liver weight (upon sacrifice). In addition, histological changes in the liver were evaluated by hematoxylin-eosin (HE) and oil red O staining. The significance of differences between groups was assessed by statistical analysis, using the

METHODSof t-test, Wilcoxon rank sum test, x2 test, F test or Fisher's test as appropriate.

RESULTSAs compared to the mice in the SC group at the corresponding time points, the mice in the HFHFr and HF groups showed significantly higher body mass and wet liver weight, as well as more extensive and robust lipid disposition in hepatic tissues as evidenced by oil red O staining. However, HE staining indicated that the HFHFr and HF groups had different degrees of macrosteatosis accompanied with intralobular inflammatory foci, with the former showing more remarkable NASH-related histological changes. Analysis at the end of week 16 showed that about 80% of the mice in the HFHFr group had developed NASH [nonalcoholic fatty liver disease (NAFLD) activity score (NAS): less than 5]. The levels of low-and high-density lipoprotein (LDL and HDL) cholesterol, as well as the levels of ALT and AST, were increased from the end of week 4 to the end of week 8 for the HFHFr and HF groups. At the end of week 16, the two groups differed in the extent of increase in total cholesterol and LDL and HDL cholesterol, with only the HFHFr group showing statistically significant changes. Specifically, at the end of week 16, the HFHFr group showed ALT levels of 108.5 +/- 93.34 U/L (F=5.099, P =0.005 vs. HF group: 44.30 +/- 35.71 U/L, HFr group: 46.70 +/- 17.95 U/L, SC group: 24.70 +/- 6.57 U/L), AST levels of 316.30 +/- 208.98 U/L (F=6.654, P=0.001 vs. HF: 132.12 +/- 75.43 U/L, HFr: 143.30 +/- 38.53 U/L, SC: 122.60 +/- 12.76 U/L), total cholesterol levels of 5.18 +/- 0.58 mmol/L (F=72: 470, P =0.000 vs. HF: 3.94 +/- 0.75 mmol/L, HFr: 2.30 +/- 0.50 mmol/L, SC: 2.02 +/- 0.24 mmol/L), HDL cholesterol levels of 3.05 +/- 0.49 mmol/L (F=25.413, P =0.000 vs. HF: 2.65 +/- 0.54 mmol/L HFr: 1.77 +/- 0.47 mmol/L, SC: 1.58 +/- 0.16 mmol/L), LDL cholesterol levels of 1.11 +/- 0.23 mmol/L (F =83.297, P =0.000 vs. HF: 0.72 +/- 0.17 mmol/L, HFr: 0.27 +/- 0.04 mmol/L, SC: 0.20 +/- 0.05 mmol/ L).

CONCLUSIONThe present study suggests that a mouse model of NASH can be successfully induced by a 16-week modified HFHFr diet.

Animals ; Diet, High-Fat ; Disease Models, Animal ; Fructose ; administration & dosage ; Male ; Mice ; Mice, Inbred C3H ; Non-alcoholic Fatty Liver Disease

Objective To evaluate the efficacy and complications of component blood transfusion combined with recombinant activated factorⅦa(rFⅦa)in treatment of severe active hemorrhage after cardiac surgery??Methods Fifty patients who suffered from severe active hemorrhage after cardiac surgery were selected from the First Affiliated Hospital of Dalian Medical University from July 2015 to May 2017??All patients were divided into GR group ( component blood transfusion combined with rFⅦa) and GA group (component blood transfusion combined with tranexamic acid) by random number table method,25 cases in each group??The changes of disseminated intravascular coagulation (DIC) were screened on admission(D1), after cessation of cardiopulmonary bypass ( D2 ), and 2 h ( D3 ), 6 h ( D4 ) and 12 h ( D5 ) after medication??The difference of activated partial thromboplastin time (APTT), international normalized ratio (INR),fibrinogen,hemoglobin and platelet of the two groups at each time point of D1,D2,D3,D4 and D5 were analyzed??Meanwhile, the postoperative drainage, postoperative blood transfusion, postoperative plasma transfusion volume, postoperative mechanical ventilation time, ICU retention time, the 30 d mortality and complications were compared between the two groups??Results There were significant differences in APTT, INR,fibrinogen,hemoglobin and platelet between the two groups ( all P＜0??05)??There was no significant difference in the indices of DIC screening between the two groups at D1, D2 and D5 time points ( all P＞0??05),but at D3 time point,APTT in GR group was significantly shorter than that in GA group((50??3 ±6??6)s vs??(60??1±6??5)s,P＝0??027),and INR in GR group at D4 time point was also significantly lower ((1??3 ± 0??3) vs??( 1??5 ± 0??3), P＝0??041)??In addition, the amount of red blood cells transfusion after treatment in GR group and GA group (( 3??2± 1??0) U vs??(4??1 ± 1??0) U,P＝0??005),the amount of fresh plasma transfusion ((303??2±98??5) ml vs??(469??6± 190??5) ml,P＝0??000),the amount of 24 h drainage after operation ((519??9±107??5) ml vs??(657??2±100??1) ml, P＝0??000) were significantly decreased,the differences were statistically significant??Conclusion Blood component transfusion combined with rFⅦa can significantly improve APTT and INR of severe active hemorrhage after cardiac surgery,at the same time,it can reduce the amount of red blood cells transfusion and plasma transfusion??

OBJECTIVE: To explore the genetic basis of cerebral palsy (CP). METHODS: A pair of twins with cerebral palsy and different phenotypes were subjected to whole genome sequencing, and other 8 children with CP were subjected to whole exome sequencing. Genetic variations were screened by a self-designed filtration process in order to explore the CP-related biological pathways and genes. RESULTS: Three biological pathways related to CP were identified, which included axon guiding, transmission across chemical synapses and protein-protein interactions at synapses, and 25 susceptibility genes for CP were identified. CONCLUSION The molecular mechanism of CP has been explored, which may provide clues for development of new treatment for CP.
Cerebral Palsy ; genetics ; Child ; Genetic Testing ; Humans ; Phenotype ; Whole Exome Sequencing ; Whole Genome Sequencing

Objective To investigate the feasibility of using a sitting aldosterone renin ratio (ARR) instead of a standing ARR to screen for primary aldosteronism (PA) in hypertensive patients. Methods From April to July 2017, 63 hypertensive patients admitted to the Department of Hypertension of Fuwai Hospital of Chinese Academy of Medical Sciences were selected as subjects by convenient sampling. During the hospitalization, the patients performed two position tests on different days. On the first day, the patients were asked to keep the standing and walking position for 2 hours, and the standing blood samples were taken. The next day, the patients were asked to keep the sitting position for 2 hours and then take the blood samples. Comparing ARR results for both body position measurements. Results A total of 62 patients successfully completed the study. There were no significant differences in peripheral blood ARR results between the sitting and standing position of patients with same age groups and genders (P＞ 0.05). Conclusions The sitting position ARR does not affect the PA test results. It is recommended to use the sitting position ARR for PA screening in clinical practice,which can help reduce the incidence of adverse reactions and interruption events.

Objective:To explore the clinical utility of liquid chromatography tandem mass spectrometry forprimary aldosteronism screening.Methods:From January to October 2019, 413 inpatients diagnosed hypertension from Fuwai Hospital of Chinese Academy of Medical Sciences were enrolled, including 60 Primary aldosteronism(PA)patients and 353 primary hypertension patients. The plasma aldosterone concentration (PAC) and renin concentration (DRC) were measured after 2 h of standing. The 24 h urine samples were collected for measurement of aldosterone using LC-MS/MS. The performance of urine aldosterone and urine aldosterone/renin ratio (UADRR) in PA screening was evaluated by ROC, and compared with PAC/DRC ratio (ADRR). Meanwhile, the efficiency of urine aldosterone in elderly patients or patients with low blood potassium or 24 h urine sodium over 200 mmol was investigated.Results:Area under the curve (AUC)of urine aldosterone was 0.725 (95 %CI 0.679-0.767), and the best cut-off was 7.13 μg/24 h, which was lower than AUC of ADRR (0.958, 95 %CI 0.934-0.975). The AUC of UADRR was 0.947 (95 %CI 0.920-0.966), the best cut-off was 1.11 (μg/24 h)/(μIU/ml), the sensitivity and specificity were 91.7% and 89.0%, respectively. There is no significant differences found with ADRR. In patients with 24 h urine sodium over 200 mmol, AUC of aldosterone was 0.834 (95 %CI 0.730-0.910) and the best cut-off was 9.31 μg/24 h. The sensitivity and specificity were 90.9% and 68.7%, respectively. For the elderly patients over 60 years old, the AUC of urinary aldosterone was 0.860 (95 %CI 0.770-0.925), and the best cut-off was 6.91 μg/24 h. The sensitivity and specificity were 84.6% and 81.3%, respectively. When admission blood potassium was less than 3.50 mmol/L, AUC of urinary aldosterone was 0.822 (95 %CI 0.684-0.917), and the best cut-off was 10.63 μg/24 h. The sensitivity and specificity were 85.7% and 66.7%, respectively. Conclusion:The detection of aldosterone in urine by LC-MS/MS can provide clinical information for PA screening, and the screening performance is better in patients with 24-hour urine sodium over 200 mmol, elderly patients or patients with low blood potassium. If combined with renin, screening efficiency was the same as that in ADRR.

Objective To investigate the co-occurrence incidence,clinical features and risk predictors of autism and intellectual disability in patients with Lennox-Gastaut syndrome (LGS).Methods Sixty-four patients with LGS were recruited in our Epilepsy Center from June 2012 to June 2018.Autism Behavior Checklist (ABC) and Childhood Autism Rating Scale (CARS) were performed to evaluate autism,while Chinese Wechsler Intelligence Scale for Children (C-WISC) and Gesell Developmental Scale were applied to estimate intelligence.The influences of different clinical factors in autism and intellectual disability were analyzed in patients with LGS.Results Among 64 patients with LGS,only three (4.7％) were diagnosed as having autism,and their average ABC and CARS scores were 80.0 and 40.0,respectively.The average ABC and CARS scores were 40.9±26.7 and 26.0±8.9 in thepatients with onset age＜one year,which were significantly higher than those in other two groups,respectively (P＜0.05).The average ABC and CARS scores in the patients accepted antiepileptic drugs (AEDs) ≥ 3 were 27.8±22.8 and 22.2±8.7,which were significantly higher than those in the patients accepted one or two kinds ofAEDs (P＜0.05).In addition,the ABC and CARS scores showed significant differences in the groups with different seizure frequency and in the groups with or without symptomatic etiologies (P＜0.05).Fifty patients (78.1％) presented different levels of intellectual disability;severe intellectual disability was the leading type,which accounted for 31.3％ (20/64);12(18.8％),7(10.9％),and 11 (17.2％) patients were with mild,moderate or profound intellectual disability,respectively.As compared with patients without intellectual disability,patients with intellectual disability had younger onset age,higher proportion of slow background activity on EEG and higher proportion of symptomatic etiologies,with significant differences (P＜0.05).Conclusion Patients are in higher risk of autism when they have earlier epilepsy onset age,higher frequency of epilepsy seizure attack,administration of AEDs ≥3 and symptomatic etiologies;early onset age is an independent risk predictor for intellectual disability of patients with LGS.

OBJECTIVE To provide reference for the diagnosis and treatment of Stevens-Johnson syndrome caused by rebamipide， and to explore the predisposing factors of Stevens-Johnson syndrome. METHODS Clinical pharmacists analyzed the treatment process of a patient with gastrointestinal diseases and evaluated the correlation between the drug used and adverse reactions， in order to determine the suspected allergenic drug causing Stevens-Johnson syndrome. The predisposing factors of patients with Stevens-Johnson syndrome were explored. RESULTS & CONCLUSIONS The suspected allergenic drugs that caused the patient to develop Stevens-Johnson syndrome included Ilaprazole enteric-coated tablets， Rebamipide tablets and Kangfuxin liquid. In summary， the suspect drug was identified as Rebamipide tablets according to the causality evaluation method of the National Center for Adverse Drug Reaction Monitoring， Naranjo’s scoring method and the algorithm of drug causality for epidermal necrolysis scoring criteria. Hypoproteinemia， competitive binding of plasma proteins between drugs， advanced age， bacterial and viral infections were the predisposing factors of Stevens-Johnson syndrome. Therefore， before using rebamipide in clinical practice， it is necessary to inquire about the patient’s allergy history in detail. During the use process， it is necessary to strengthen the patient’s medication monitoring and be alert to the occurrence of serious adverse reactions. If any abnormalities are found， the medication should be stopped immediately and symptomatic treatment should be given as soon as possible to ensure the safety and effectiveness of the patient’s medication.

Objective: To evaluate the effect of 3D-printed oval root canal preparation by using small field-of-view cone beam CT (CBCT) combined with computer-aided technologies. Methods: An extracted tooth with a suitable single oval canal was selected by small field-of-view CBCT scanning. Three-dimensional (3D)-printed resin teeth were obtained based on the CBCT data after data conversion and processing. 50 general dentists were selected to prepare the oval root canal of the resin teeth with X-Gold rotary Ni-Ti instruments. Small field-of-view CBCT was applied to scan the oval root canals before and after preparation. Then, computer-aided technologies were used to calculate and compare these CBCT data, analyzing the effects of oval root canal cleaning as well as the root canal deviation by 3D reconstruction and rendering of the images. Results : Among the 50 cases, the mean unprepared area of the oval root canal wall was（56.20 ± 11.91）% and the mean maximum deviation distance of the root canal was（0.140 ± 0.041）mm. There was no correlation between root canal cleanliness and deviation (r=0.18, P=0.212). Conclusion Combined with small field-of-view CBCT and computer-aided technology, we can effectively quantitatively evaluate the root canal preparation effect.