The solvent sublation technique was applied for the separation and enrichment of L-Arg using dodecylbenzene sulfonic (DBSA) as the surfactant, di (2-ethylhexyl) phosphoric acid (P204) as the extractant and n-heptane as the organic solution. The solvent sublation was compared with the floatation complexation extraction, foam floatation and solvent extraction. The experimental results showed that enrichment ratio of 16.2 and removal rate of 97.2% to L-Arg were obtained by the solvent sublation under the conditions of room temperature, 0.09 g/L L-Arg aqueous solution 250 mL, DBSA concentration 0.15 g/L, the initial pH 7.0, volume of n-heptane 10 mL, volume of P204 4.5 mL, gas flow rate of 200 mL/min. The study of the kinetics indicated that the solvent sublation process could be divided into three stages distinctly. The processes of the first stage and the second stage followed the first order kinetics equation; the process of the third stage followed the zero order kinetics equation. The separation mechanism of solvent sublation was also discussed.

Objective To study the changes of P-selectin and E-selectin in pediatric patients with critical illness ,and analyze their relationship with the severity and prognosis of diseases.Methods Forprospective study,42 critically ill patients admitted in pediatric intensive care unit (PICU ) from September,2012 to March,2013 as critically ill group were enrolled,and blood specimens were collected with 24 hours after admission.Another 42 cases blood samples were collected from children's physical examination as control group.The severity of the critically ill patients were evaluated by Pediatric Critical illness Score (PICS)and Pediatric risk of score mortality (PRISM)-III.The levels of serum P-selectin and serum E-selectin were measured by double antibody sandwich enzyme-linked immunoassay (ABC-ELISA). Results P-selectin and E-selectin in control group children and critically ill patients group were (37.23 ± 8.99)ng/mL,(36.24 ±17.82)ng/mL,and (107.24 ±35.53)ng/mL,(114.93 ±40.17)ng/mL, respectively.There were statistical differences between two groups (P=0.000).The levels of P-selectin and E-selectin in acute phase were higher than that of levels in recovery phase in critically ill group (P =0.000).Negative correlation was observed between P-selectin concentration and the PCIS score (r =-0.673,P=0.000),as well as E-selectin (r=-0.548,P=0.000).P-selectin level and E-selectin level based upon PRISMⅢ≥10 group were significantly higher than they in PRISMⅢ <10 group (P=0.003,P=0.014).In critically ill children,the differences in P-selectin,E-selectin were significant higher in death patients (P=0.003;P =0.000).Compared with the non-sepsis illness group,the level of P-selectin and E-selectin in the severe sepsis patients were significantly higher (P =0.04,P =0.025 ). Conclusions The levels of P-selectin and E-selectin are closely related to the severity and prognosis in critically ill children.Measuring the level of P-selectin and E-selectin could be used as a judegment the severity and to understand pathological physiological process.

Objective To investigate the changes of epithelial neutrophil activating peptide-78 (ENA-78) in the serum of patients with critical illness,and to analyze the relationship between the severity and prognosis.Methods Prospective case-control study was performed,and 42 cases of critically ill patients admitted to Pediatric Intensive Care Unit,Children's Hospital Affiliated to Shanghai Jiaotong University from Sep.to Nov.2013 were selected as critically ill group,blood specimens were collected within 24 hours and 7 days after their admission.Another 42 cases of blood samples were collected during physical examinations in this hospital as control group.The severity of critically ill patients were graded by Pediatric Critical Illness Score (PICS) and Pediatric Risk of Score Mortality (PRISM) Ⅲ,and the serum ENA-78 was measured by double antibody sandwich enzyme-linked immunoassay.Results 1.The level of ENA-78 in the control group was (0.44 ± 0.28) ng/L; ENA-78 in acute phase and recovery phase of critically ill group were (2.85 ± 0.89)ng/L and (1.00 ± 0.64)ng/L,respectively,there were statistical differences between control group and critically ill group,acute phase group and recovery phase group (all P =0.000).2.The negative correlation was observed between ENA-78 concentration and PCIS score(r =-0.724,P =0.000).ENA-78 in PRISM Ⅲ ≥ 10 group was significantly higher than that in PRISM Ⅲ＜ 10 group(P =0.000).The ENA-78 between death group and the survival group was significantly different(P =0.000).3.ENA-78 in patients with severe infection was higher than that in the non-infectious cases(P =0.000).4.With the organ dysfunction expanded ENA-78 rose accordingly,and the difference was statistically significant (P =0.000).Conclusions The level of ENA-78 is different in critically ill patients in children.It can provide reference of assessing the severity of disease and predicting prognosis by determing the ENA-78 level.

Objective To investigate the correlation between tumor perfusion parameters and tumor volume and Child-Pugh classification in CT of hepatocellular carcinoma (HCC).Methods Fifty-six patients with hepatocellular carcinoma were selected to perform CT perfusion imaging.The parameters of the total tumor perfusion such as hepatic artery perfusion (HAP),portal vein perfusion (PVP)and hepatic perfusion index (HAPI)were calculated according to the degree of hepatic encephalopathy,albumin,bilirubin, clotting time,ascites for liver Child-Pugh classification.The relationship between the tumor or peritumoral perfusion parameters with tumor volume and Child-Pugh classification were analyzed.Results (1)There was no correlation between tumor or peritumoral perfusion with the tumor volume.(2)The difference of HAP,PVP and HAPI between the different Child-Pugh classification groups was statistically significant (P<0.000 1).(3)With the reduction of Child-Pugh classification,the tumor body HAP and HAPI values gradually decreased, while the PVP value increased gradually.Conclusion There is no correlation between the tumor volume of hepatocellular carcinoma with total tumor perfusion parameters.The differences in perfusion measurements between different Child-Pugh classification can intuitively and quantitatively reflect the reserve function of the liver.

Objective:To establish a predictive model of moderate to severe pain in patients with hepatocellular carcinoma (HCC) undergoing transcatheter arterial chemoembolization (TACE).Methods:264 patients with HCC who underwent TACE operation in Southern Medical University from January 2017 to April 2018 were selected as the modeling set. The pain was assessed by numeric rating scales. The patients were divided into pain group ( n=96) and non-pain group ( n=168) according to whether moderate to severe pain occurred within 24 hours after the operation. Binary Logistic regression analysis were performed for variables that were statistically significant in the univariate analyses. The predictive nomogram was constructed and the internal validation was performed. In addition, 87 patients with HCC who underwent TACE operation from January 2020 to June 2020 were selected as the validation set for external validation. Results:In the modeling set, 96 patients (36.36%) had moderate to severe pain within 24 hours after TACE operation in 264 patients with HCC, and the dosage of morphine intramuscularly injected within 24 hours was 1015 mg, with an average of 10.57 mg per patient. Multivariate Logistic regression analysis showed that preoperative pain, the distance between the tumor and capsule ≤2 cm, high prothrombin activity, dosage of lipiodol>10 ml, and several thromboembolic tumors were independent risk factors for moderate to severe pain after TACE ( P<0.05). Age>50 was the protective factor of moderate to severe pain after TACE ( P<0.05). The area under ROC curve was 0.799 (95% CI: 0.745-0.853) in the modeling set. The area under Roc curve for internal validation and external validation were 0.780 and 0.788, respectively. The calibration curves showed satisfactory agreements between the model predicted probability and the actually observed probability. Conclusion:The predictive model of moderate to severe pain after TACE was established in this study has good differentiation and accuracy, it has certain guiding significance for predicting the high-risk group of moderate to severe pain after TACE operation and formulating the targeted prevention strategy.

Objective To investigate the effect of long-term high-fat diet on cognitive function and hippocampus neurons ultrastructure in obese rats.Methods Forty SD rats were randomly assigned to a high fat diet (HFD) group and a common diet (CD) group.Meanwhile,HFD-induced obese rat model were established.The spatial learning and memory were measured by the Morris water maze,and the neurons ultrastructural changes in rat hippocampus CA1 region at the corresponding period were observed by transmission electron microscopy.Results The average weight of rats was 25％,28％,and 22％ higher in the HFD group than in the CD group at the 12,16,and 20 weeks,respectively;the Lee's indexes were 6％,4％,and 8％ higher;the average swimming latency were 52％,44％,and 40％ longer;the average swimming distance were 85％,45％,and 51％ longer;the average swimming speed were 57％,34％,and 18％ higher;the duration of staying in the target quadrant were 32％,54％,and 63％ shorter;and the average times of crossing the plate form were 30％,34％,and 34％ shorter,respectively (all P ＜0.001).In comparison of ultrastructure in hippocampus CA1 region of rats at corresponding time points,the amounts of degenerated and necrosis neurons,of the deformed and vacuolar mitochondria,and of the less rough endoplasmic reticulum were significantly more at 12,16,and 20 weeks in the HFD group than in the CD group.Conclusions Long-term HFD-induced obesity damages the structure of neurons in the hippocampus,impairs spatial learning and memory function,and accelerates cognitive aging in rats.

Objective:To explore the effects and the mechanism of metformin combined with celecoxib on the proliferation and apoptosis of hepatoma HepG2 and Huh7 cells.Methods:Hepatoma cells HepG2 and Huh7 were divided into control group, metformin group, celecoxib group and combination medication group, CCK-8 assay was used to detect cell proliferation; Hoechst33258 staining method was used to investigate the cell apoptosis; wound healing test was used to detect cells migration ability; Transwell invasion chamber test was used to detect cell invasion ability; Western blotting was used to detect the expression of AMPK, PI3K, Akt, mTOR.Results:After metformin and celecoxib treatment, HepG2 and Huh7 cells were gradually contracted, disintegrated and more apoptotic cells were noticed, and cell proliferation was significantly inhibited. The wound healing test results showed that the cell migration was significantly decreased ( P<0.05) under metformin and celecoxib treatment. The results of the transwell invasion chamber test showed that the metformin and celecoxib treatment inhibited the invasion of HepG2 and Huh7 cells ( P<0.05). The expression levels of AKT, AMPK, and mTOR were decreased in HepG2 cells in the combinational treatment group, and the expression level of PI3K was decreased and then increased; the expression levels of AKT, AMPK, PI3K, and mTOR in Huh7 cells were decreased. Conclusions:Metformin can cooperate with celecoxib to enhance the inhibitory effect on the proliferation, migration and invasion of HepG2 and Huh7 cells. The mechanism may be related to the inhibition of the expression of mTOR signaling pathway.

Objective To explore the utility of CT for the evaluation of abdominal aortic calcification(AAC)in chronic kidney disease(CKD).Methods Laboratory examination indexes and abdominal plain CT imaging of 132 CKD stage 3-5 patients were analyzed retrospectively.The ACC score was evaluated according to quantitative method,the risk factors related to AAC were analyzed.Results In correlation analyses,AAC score was positively correlated with creatinine(Cr)level,CO2 combining power(P＜0.05),and negatively corrected with albumin(ALB).Logistic regression analysis showed that Cr level were risk factors for AAC,and ALB were protective factors.Conclusion Based on abdominal plain CT images,Cr level,CO2 combining power and ALB level are related to AAC.Quantitative method based on CT has a high application value in evaluating AAC.

Objective:To explore whether thyroxine (T 4) could promote differentiated thyroid cancer (DTC) progression by binding to integrin α vβ 3in vitro and its downstream mechanism. Methods:Papillary thyroid cancer cell lines TPC-1, K1 and follicular thyroid cancer (FTC) cell line FTC133 were cultured in vitro, and the expressions of integrin α vβ 3 in those 3 DTC cell lines were determined with immunofluorescence and flow cytometry analysis. After the treatment of T 4, tetraiodo thyroacetic acid (Tetrac) and Arg-Gly-Asp (RGD) peptide alone or in combination, the proliferation and metastatic potential of DTC cell lines were detected by cell counting kit-8 (CCK-8), Transwell migration and invasion assays. The small interfering RNA (siRNA) transfection was used to verify whether integrin α v or β 3 subunit knockdown could reverse the effect of T 4 on DTC cells. The expression levels of downstream signaling proteins phosphorylated extracellular signal-regulated kinase (p-ERK)1/2 and total extracellular signal-regulated kinase (ERK)1/2 were detected by Western blot. The effects of mitogen-activated protein kinase kinase (MEK)1/2 inhibitor (GSK1120212) on the proliferation, migration and invasion of T 4-treated cells were detected. One-way analysis of variance and Tukey test were used for data analysis. Results:The integrin α vβ 3 expressions in TPC-1, K1 and FTC133 cells were all positive, with the relative mean fluorescence intensity (MFI) of 61.93±18.61, 16.89±2.43 and 32.36±0.83, and the percentages of positive cells of (94.38±1.30)%, (74.11±3.87)% and (50.67±1.78)%, respectively ( F values: 13.36 and 217.30, P=0.006 and P<0.001). Compared with control group, the proliferation, migration and invasion in the three DTC cell lines treated with T 4 were significantly enhanced (96 h, F values: 62.67-297.50, q values: 13.15-20.73, all P<0.001). T 4-induced cell proliferation, migration and invasion were markedly reversed by Tetrac or RGD (96 h, q values: 8.61-17.54, all P<0.001). T 4-induced cell proliferation, migration and invasion were also significantly inhibited by the knockdown of integrin α v or β 3 subunit (72 h, F values: 7.75-70.98, q values: 4.77-15.21, all P<0.05). Western blot results showed that the phosphorylation levels of ERK1/2 in DTC cells were significantly increased by T 4 treatment, and the T 4-induced activation of ERK1/2 signaling pathway could be blocked by Tetrac, RGD, integrin α v or β 3 subunit knockdown. T 4-induced cell proliferation, migration and invasion were significantly reversed by GSK1120212 (96 h, F values: 47.53-151.40, q values: 10.32-16.65, all P<0.001). Conclusion:T 4 can promote cell proliferation and metastasis of DTC cells by binding to integrin α vβ 3 and activating the ERK1/2 pathway.

OBJECTIVE：To establish UPLC fin gerprint of 32 compatible herb pairs with Polygonum multiflorum as the core ， and to conduct multivariate statistical analysis. METHODS ：UPLC method was adopted. Using P. multiflorum and single decoction pieces of compatible herb as reference ，UPLC fingerprints of 32 compatible herb pairs of P. multiflorum were drawn. Common peaks were confirmed by relative retention time and UV absorption spectrum. Non-supervised PCA and supervised OPLS-DA were conducted by using SPSS 19.0 software and SIMCA 13.0 software. RESULTS ：There were totally 12 common peaks in UPLC fingerprints of 32 compatible herb pairs of P. multiflorum . The results of non-supervised PCA showed that the accumulative variance contribution rate of primary 6 principal components was 84.633％. The results of cluster analysis of PCA comprehensive score showed that single decoction piece of P. multiflorum ，compatible herb pairs of P. multiflorum with Lycium barbarum ，Rehmannia glutinosa，Paeonia lactiflora ，Codonopsis pilosula ，Eclipta prostrate ，Angelica sinensis ，Glycyrrhiza uralensis ，Astragalus membranaceus and Ophiopogon japonicus were clustered into one category ；others were clustered into one category. Results of supervised OPLS-DA analysis showed that eigen values of 4 principal components were 2.32，2.61，1.58 and 0.90，respectively. There were differences in the contents of 12 common components in the compatibility of P. multiflorum with tonic medicine and non-tonic medicine. The changes of the content of the components after compatibility with tonic medicine were similar. Common peak 7，4，6，3 were main reasons for the differences （variable importance projection value were all higher than 1）. CONCLUSIONS：Established fingerprint is simple in operation ，and can be combined with multivariate statistical analysis to evaluate the content changes of common components of 32 compatible herb pairs with P. multiflorum as the core.