Although the functions of metabolic enzymes and nuclear receptors in controlling physiological homeostasis have been established, their crosstalk in modulating metabolic disease has not been explored. Genetic ablation of the xenobiotic-metabolizing cytochrome P450 enzyme CYP2E1 in mice markedly induced adipose browning and increased energy expenditure to improve obesity. CYP2E1 deficiency activated the expression of hepatic peroxisome proliferator-activated receptor alpha (PPARα) target genes, including fibroblast growth factor (FGF) 21, that upon release from the liver, enhanced adipose browning and energy expenditure to decrease obesity. Nineteen metabolites were increased in Cyp2e1-null mice as revealed by global untargeted metabolomics, among which four compounds, lysophosphatidylcholine and three polyunsaturated fatty acids were found to be directly metabolized by CYP2E1 and to serve as PPARα agonists, thus explaining how CYP2E1 deficiency causes hepatic PPARα activation through increasing cellular levels of endogenous PPARα agonists. Translationally, a CYP2E1 inhibitor was found to activate the PPARα-FGF21-beige adipose axis and decrease obesity in wild-type mice, but not in liver-specific Ppara-null mice. The present results establish a metabolic crosstalk between PPARα and CYP2E1 that supports the potential for a novel anti-obesity strategy of activating adipose tissue browning by targeting the CYP2E1 to modulate endogenous metabolites beyond its canonical role in xenobiotic-metabolism.

Objective: Symptomatic intracranial hemorrhage (sICH) is one of the severe complications of ischemic stroke thrombolysis. Several prognostic scales have been developed to predict the risk of sICH. The performance of seven scales was compared in a single center cohort. Methods: Data of patients with consecutive ischemic stroke who received 0.9 mg/kg intravenous recombinant tissue plasminogen activator (rt-PA) thrombolysis within 4.5 h time window from stroke onset were collected. Seven scales that can provide an estimate of risk of sICH were identified and evaluated: Hemorrhage After Thrombolysis (HAT), blood Sugar, Early infarct signs, (hyper) Dense cerebral artery sign, Age, National Institutes of Health (NIH) Stroke Scale (SEDAN), Stroke Prognostication using Age and NIH Stroke Scale (SPAN)-100, Safe Implementation of Thrombolysis in Stroke (SITS), Total Health Risks In Vascular Events (THRIVE), Glucose at presentation, Race (Asia), Age, Sex (male), systolic blood Pressure at presentation, and Severity of stroke at presentation (NIH Stroke Scale; GRASPS) and Multicenter Stroke Survey (MSS). The area under the receiver operating characteristic curve (AUROC) was calculated and Logistic regression and the Hosmer-Lemeshow test were also performed. Results: The current study included 293 patients, of whom 7.85% (23/293) had sICH by National Institute of Neurological Disorders and Stroke (SICH_NINDS), 5.46% (16/293) by Europe Cooperative Acute Stroke Study Ⅱ (SICH_ECASSⅡ) and 4.44% (13/293) by Safe Implementation of Thrombolysis in Stroke (SICH_SITS) criteria. SEDAN had the highest AUROC for predicting sICH: sICH_NINDS: AUROC=0.843, OR=3.167, 95%CI 2.106-4.762, P<0.01; sICH_ECASSⅡ: AUROC=0.797, OR=2.509, 95%CI 1.652-3.812, P<0.01; sICH_SITS: AUROC=0.784, OR=2.172, 95%CI 1.405-3.357, P<0.01. And SPAN-100 had the lowest AUROC among all the seven scales and was only associated with risk of SICH_NINDS in regression analysis. Furthermore, when sub-grouped the cohort into anterior circulation infarction and posterior circulation infarction, regression analysis suggested that all the seven scales were however not associated with sICH risk in patients with posterior circulation infarction. Conclusions SEDAN constantly had the highest predictive power, SPAN-100 had the worst. The seven scales studied could not predict sICH in posterior circulation infarction.

Objective To explore the role of CORM-3 in alleviating cognitive dysfunction and cortical neuronal pyroptosis of rats exposed to hemorrhage shock and resuscitation. Methods One hundred and sixty-eight male SD rats, weighting 350-400 g, in accordance with random number table, were divided into 4 groups (n=42):sham-operated group, hemorrhage shock and resuscitation (H group), hemorrhage shock and resuscitation plus CORM-3 (CO group), hemorrhage shock and resuscitation plus iCORM-3 (ICO group). The rat hemorrhagic shock resuscitation models were established in H, CO and ICO groups:bleeding from femoral vein was performed to achieve mean arterial pressure of 25-35 mmHg (1 mmHg=0.133 kPa) for 60 min;and then, the collected blood was returned to the body within 15 min to reach the initial blood pressure level as resuscitation, and normal saline was injected if necessary. The rats in CO group were injected CORM-3 (4 mg/kg) via femoral vein after resuscitation, the rats in ICO group were injected iCORM-3 (4 mg/kg), and rats in the sham-operate group and H group were only injected equal amount of normal saline containing DMSO. Rats were sacrificed for cortex 12 h after end of resuscitation;CO content was assessed by gas chromatograph assay; Western blotting was used to detect the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and BTB-CNC homology 1 (Bach1) in the nuclear, and heme oxygenase-1 (HO-1), interleukin (IL)-1β and IL-18 in cytosol; and neuronal pyroptosis rate was detected by cleaved caspase-1-Cy3/neuron-specific nucleoprotein (NeuN)-FITC/DAPI. Thirty d after resuscitation, open field test was used to assess the cognitive ability of the rats. Results At 12 h after resuscitation, as compared with sham-operated group, rats in the H, CO and ICO groups had significantly increased CO content, neuronal pyroptosis, ratio of Nrf2/Bach1, and expressions of HO-1, IL-1β and IL-18, statistically longer time spending in the central square, significantly smaller times crossing the grid and times standing on the back legs (P<0.05). As compared with H and ICO group, CO group had significantly increased CO content, ratio of Nrf2/Bach1, and HO-1 expression, times crossing the grid, times standing on the back legs, but significantly decreased neuronal pyroptosis, expressions of IL-1β and IL-18, and time spending in the central square (P<0.05). Conclusion CORM-3 can reduce the neuronal pyroptosis rate and alleviate cognitive dysfunction in rats with hemorrhagic shock and resuscitation, whose mechanism may be related to the increase of HO-1 expression after up-regulation of Nrf2/Bach1 ratio.

To develop a simple and easy promotion risk score to identify individuals with undiagnosed papillary thyroid carcinoma (PTC), and further to compare the diagnostic efficiency of PTC risk sore with fine needle aspiration cytodiagnosis (FNAC). in order to optimize the screening process of PTC. A sample of 1003 individuals aged 11-82 years underwent a surgical treatment of thyroid nodule participated in the study. The risk score was developed by stepwise backward multiple logistic regression. And using the receiver operating characteristic (ROC) curve to evaluate the diagnostic efficacy, the best diagnostic cut-off point and the risk stratification of malignant. Compare the sensitivity, specificity, accuracy, and area under curve ( AUC) of PTC risk score, FNAC and their combined diagnosis to judge their diagnostic efficiency. The risk score included age, TSH, nodule morphology and boundary by palpation, nodule characteristics, shape, boundary, calcification and blood flow signal by ultrasound. Its AUC= 0. 815, 6 point was the best cutoff point to differentiate benign from malignant thyroid nodules, and risk stratification of thyroid carcinoma were divided into four levels: very high risk group (score ≥ 9 points), high risk group ( score were 5-8 points), moderate risk group ( score were 3 ~ 4 points), low risk group ( score ≤ 2 points). The sensitivity, specificity, and accuracy of FNAC were 86. 3% , 90. 0% , and 87. 0% respectively, AUC=0. 891, while the sensitivity, specificity, and accuracy of PTC risk rating scale were 83. 8% , 70. 0% , and 81. 0%respectively, AUC = 0. 822. The sensitivity, specificity, and accuracy of their combined diagnosis were 97. 5% , 85. 0% , and 95. 0% , AUC=0. 965. This risk score can be used as a screening method before FNAC. If combined with FNAC, it may improve the diagnostic efficacy of PTC, and thereby possibly minimizing the unnecessary invasive examination and surgical treatment for patients with thyroid nodules and reducing personal costs.

Objective:To treat the mixed and deeper infantile hemangioma with ladder dosage propranolol, and to explore its efficacy and safety.Methods:A total of 98 infants with hemangioma were treated by ladder treatment of propranolol.Before treatment,comprehensive assessments of electrocardiogram(ECG),heart color ultrasound, blood glucose,liver function,kidney function,myocardial enzymes and blood routine were conducted.After excluding contraindications,the dose of propranolol incrementally doubled from 0.5 mg·kg-1·d-1 to 4.0 mg·kg-1·d-1.Propranolol was taken 3 times a day.Before and after medication for 1 and 2 h,ECG was monitored.The changes of tumor size,texture,color and other changes or an onset of adverse reactions were dynamicly observed.The infants were visited every month.The efficacy was evaluated using Achauer system.Results: After medication,98 cases had different degrees of color changes or tumor consistency softening.After the dosage of propranolol was increased to 4.0 mg·kg-1·d-1,the change of tumor was the fastest.According to the 4-grade method, there were 84 cases(85.71%) as gradeⅣ (excellent),2 cases (2.04%) as grade Ⅲ (good),4 cases (4.08%) as gradeⅡ (medium)and 8 cases (8.16%) as gradeⅠ (poor).The curative effect of mixed hemangioma was better than that of deeper hemangioma(P<0.05).The recovery time of 74 cases of hemangiomas was 6 months.The major adverse reactions were heart rate decline(5/98,5.10%),drowsiness(3/98,3.06%),diarrhea(7/98,7.14%),loss of appetite (1/98,1.02%), and convulsions (2/98,2.04%).After treatment,all adverse reactions disappeared.Two months after drug withdrawal there were 4 cases of recurrence,and they were continously treated with propranolol.Conclusion: The efficacy of oral ladder dosage propranolol in treatment of mixed and deeper infantile hemangioma is increased significantly and there are no significant adverse reactions.

Objective To investigate the expression level and clinical significance of long non-coding RNA(LncRNA) growth arrest specific gene-antisense 1(GAS8-AS1) in papillary thyroid microcarcinoma(PTMC) patients. Methods We investigated the expression profile of GAS8-AS1 in tissue samples of patients with PTMC as well as nodular goiter(NG) by quantitative real-time polymerase chain reaction(RT-qPCR). Results GAS8-AS1 in cancer tissue was down-regulated in PTMC patients compared with adjacent thyroid tissue and NG samples(P<0.05). Lower level of GAS8-AS1 was also correlated with central cervical lymph node metastasis(CLNM, P<0.05). The area under the ROC curve for GAS8-AS1 was up to 0.717 3 in CLNM prediction(P<0.05). Conclusion GAS8-AS1 may act as a potential biomarker for PTC diagnosis and CLNM prediction.

Objective To establish a risk rating scale for central lymph node metastasis in papillary thyroid carcinoma and make risk stratification.Methods Data of 502 patients with PTC who were treated in Beijing Shijitan Hospital between January 2010 and June 2015 were retrospectively analyzed.The independent predictors for CLNM were found.Then a risk rating scale was established and stratification risk was made.The diagnostic value of the risk rating scale in predicting CLNM was evaluated.Data of 100 patients with PTC who were treated in Beijing Shijitan Hospital between July 2016 and June 2016 were used to validate the risk rating scale.Results A cutoff value of 5 points was found to be the best prediction for CLNM,with the sensitivity and specificity were 73.8％ and 70.2 ％.We definite score ≤ 4.5 as low risk for CLNM,as well as score from 5 to 7 as middle risk,score ≥ 7.5 as high risk.The other data of 100 patients was used to validate the risk rating scale.The sensitivity and specificity were 79.5％ and 78.7 ％ respectively.The positive predictive value and the negative predictive value were 70.5％ and 85.7％ respectively.Conclusions The risk rating scale provide a convenient,intuitive and quantized method to predict CLNM,which is helpful to select suitable surgical strategy and reduce operative complications.

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective Inflammatory response is an important part in pathological change of traumatic arthritis(TA).Studies show that hydrogen has antioxidate and anti-inflammatory properties, however, there are few reports on treating TA with it.So this research aimed to investigate curative effects of curing traumatic arthritis in rabbits with intraperitoneal injection of saturated hydrogen saline solution . Methods A total of 26 New Zealand white rabbits were divided into three groups:normal control group (n=6), model experimental group (n=10), and model control group (n=10).Rabbits in model experimental group were intraperitoneally injected with 8 ml /kg saturated hy-drogen saline, once every three day;equivalent saline was injected in rabbits of model control group;while no treatment was done on normal control group.The treatment lasted for 6 weeks.Before and after the treatment, the serum and joint fluid of the rabbits in the three groups were respectively taken to determine the content of nitric oxide (NO) and hyaluronic acid (HA), which was used to evaluate therapeutic effect in combination with the behavioral performance of the rabbits . Results Significant improvement in behavior was found in model ex-perimental group after the treatment (P<0.05).No statistical differences were found in knee swelling scores of the three groupsbefore the treatment(P>0.05), but model experimental group was superior to model control group in knee swelling scores after the treatment (P<0.05).The contents of NO both in serum ([79.58 ±13.46] vs [76.23 ±12.15]) and joint fluid ([89.45 ±14.98] vs [80.36 ±12.78]) in two model groups increased before the treatment, which was of statistical difference (P<0.05), but the NO content decreased dramatically in model experimental group after the treatment ([436.82 ±60.91] vs [340.21 ±40.57],P<0.05), and no obvious changes were observed in model control group (P>0.05).The content of serum HA in model experimental group declined dramatically (P<0.05) and the joint fluid increased after the treatment ([1.72 ±0.37] vs [2.47 ±0.62],P<0.05), while no significant changes were found in model control group before and after the treatment. Conclusion The intraperitoneal injec-tion of saturated hydrogen saline may be effective in the treatment of trau-matic arthritis.

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.