Objective To examine the influence of LEV adjunctive treatment on cognitive function and life quality in RE school-age children. Methods Fifty-five cases of RE children were selected from our hospital from June 2013 to December 2015. Based on original treatment all children were added with LEV treatment for 16 weeks. The initial dose was 8~10 mg/(kg·d),and gradually increased to 50 mg/(kg·d). maintenance dose was 30 mg/(kg·d). Adverse drug reac?tions were recorded and clinical efficacies were determined after treatment. Wechsler Intelligence Scale for Children re?vised in China (WISC-CR) and quality of life in children with epilepsy (QOLCE) was used to evaluate children's cogni?tive function and children's life quality, respectvely. Results Average epilepsy attack frequencies after treatment (3.8± 1.3 vs. 6.6±2.3) times/month decreased significantly compared with before treatment (P<0.05). Clinical control ratio, ex?cellent ratio, effective ratio and ineffective ratio after treatment were 9.1%, 36.4%, 43.6%, 36.4%respectively and total effective ratio was 89.1%. The arithmetic score (10.9 ± 2.6 vs. 9.2 ± 2.1) and mapping score (15.1 ± 3.9 vs. 13.8 ± 3.3) in WISC- CR evaluation increased significantly (P<0.05). Life quality score (65.7 ± 5.7 vs. 62.8 ± 4.9) , cognitive function score (60.0 ± 5.7 vs. 57.4 ± 6.2) and social function score (65.0 ± 6.3 vs. 62.5 ± 5.5) after treatment in QOLCE evaluation in?creased significantly (P<0.05). During medication, the incidences of dizziness, fatigue, lethargy, irritability, euphoria and transient aminotransferase elevation were 12.7%, 9.1%, 20.0%, 9.1%, 3.6%and 3.6%respectively. Sixteen week medica?tion retention rate was 96.4%. Conclusion LEV adjunctive treatment can significantly reduce epilepsy attacks and im?prove cognitive function and life quality in RE children, but attention should be paid to LEV's influence on mental behavior.

Objective To investigate the bioavailability and pharmacokinetics of silybin A and silybin B in rats,respectively.Methods Following iv and ig administration of silybin to 20 Wistar rats,the plasma samples were collected at different time points up to 12 h.Sample pretreatment was involved in one-step protein precipitation with acetonitrile.Silybin A and silybin B were simultaneously determined by LC-MS/MS.Results After ig dosing silybin 28,56,and 112 mg/kg to rats,the t1/2β values were 5.48,5.08,and 5.73 h for silybin A,and 4.56,4.12,and 5.53 h for silybin B; The Cmax were 674.3,1349.4,and 2042.5 ng/mL for silybin A,and 671.0,1365.4,and 2066.2 ng/mL for silybin B; The Tmax were 0.20,0.23,and 0.20 h for silybin A,and 0.20,0.23,and 0.20 h for silybin B; The AUC were 454.4,845.9,and 1219.5 h·ng/mL for silybin A,and 432.0,817.1,and 1153.6 h·ng/mL for silybin B.The absolute bioavailabilities of silybin A and silybin B were 2.86％ and 1.93％,respectively.Conclusion Silybin A and silybin B have very low bioavailability after ig administration,and there is no significant difference in the pharmacokinetic parameters between silybin A and silybin B,which indicates that the two diastereoisomers have similar pharmacokinetic behavior in rats.

ObjectiveTo evaluate the gastroprotective activity of ascaridole.MethodsThe gastroprotective effect of ascaridole was evaluated on ulcer healing in rats with acetic acid-induced chronic gastric ulcer,pylorus ligation- and Aspirininduced gastric ulcer.Ascaridole was ig administered with the dosages of 10 and 20 mg/kg once daily for 7 d.Results Ascaridole showed the significant anti-ulcer effects.In acetic acid-induced gastric ulcer rats,the ulcer areas after 10 and 20 mg/kg of ascaridole treatment were (65.1 ± 20.0) and (50.6 ± 11.0) mm2,respectively,which were significant lower (P ＜ 0.01) than that of the control group [(116.7 ± 35.8) mm2].For pylorus ligation model,ascaridole showed a gastric ulcer healing effect in a dose-dependent manner.Ascaridole at the dose of 20 mg/kg showed 50％ ulcer protection and had a significant (P ＜ 0.05) gastroprotective activity since it decreased the total acidity and pepsin activity.Compared to the control group,the two dosages of ascaridole showed the significant reduction (P ＜ 0.05) in the ulcer index on Aspirin-induced ulcer.ConclusionThis study provides evidence that ascaridole shows potential efficacy on the healing of gastric ulcers induced by acetic acid,Aspirin,and pylorus ligation.

Objective To evaluate the diagnostic efficiency of deep fungal infection by detecting the serum galactomannan ( GM) and bronchoalveolar lavage fluid ( BALF) GM, serum G test and fungal culture of BALF in patients with suspected invasive fungal infection ( IFI) in lungs.Methods A retrospective analysis was performed of the results of serum /BALF GM test ,serum G test and BALF culture from 148 patients with suspected pulmonary IFI .The indexes involved sensitivity , specificity , positive predictive value , negative predictive value , as well as diagnostic capacity for deep fungal infection with separated or combined tests .Results Among the 148 cases, 48 cases were clinically diagnosed with IFI and the rest were excluded.Among the 48 IFI cases, 3 cases were positive in serum GM test , 25 cases were positive in BALF GM test , 31 cases were positive in G test and 30 cases were positive in fungal culture .The combined detection showed a sensitivity of 91.6%,specificity of 70.0%, positive predictive value of 59.5% and negative predictive value of 94.6%.Conclusion The combination of GM/G tests and fungal culture can significantly improve the clinical diagnostic efficiency of pulmonary IFI .

Objective Survivin is overexpressed in gastric cancer. However it not expressed in normal gastric mucosa. The expression of survivin is tightly related to the prognosis of gastric cancer.By gene reconstruction we generated pcDNA3 survivin mutant(Cys84Ala) plasmid, and observed its effect on the gastric carcinoma cell lines. Methods The survivin mRNA and protein expression levels were determined by reverse transcription polymerase chain reaction(RT PCR) analysis,Western blot and immunohistochemical staining respectively . Flowcytometry and acridine orange staning were employed to detect apoptosis. Results Overexpression of survivin mRNA and protein were detected in the gastric cancer cell lines. Inhibition of survivin by survivin mutant cDNA induced apoptosis,activated caspase 3 activity,cleaved PARP and promoted cytochrome C releasing in gastric cancer cells,and effectively sensitized gastric cancer cells to chemotherapeutic agents. Conclusion Inhibition of survivin may induce apoptosis in gastic cancer and sensitize gastric cancer cells to chemotherapeutic agents.Survivin targeted therapeutic protocol may potentially benefit gastric cancer therapy.

Objective To investigate the possible signal pathway of the apoptosis in gastric cancer cell lines(SGC 7901 and MKN 45)induced by arsenic trioxide. Methods TUNEL method was used to observe the influence of calcium antagonist, inhibitors of protein kinase C (PKC) and protein tyrosine kinase(PTK) on the apoptosis of gastric cancer cells induced by arsenic trioxide. Levels of cAMP, PKC and PTK were detected before and after the treatment with arsenic trioxide. Results Both PKC and PTK inhibitors could induce apoptosis of gastric cancer cell lines, also both of them had a cooperative action with arsenic trioxide in inducing apoptosis of gastric cancer cells, while calcium antagonist had no any effect on the apoptosis of gastric cancer cell lines. PKC and PTK levels decreased but cAMP level increased during the apoptosis of gastric cancer cells induced by arsenic trioxide ( P

Objective　To investigate whether the interaction of hypertension and diabetes aggravates endothelial dysfunction and leads to smooth muscle dysfunction. Methods　Noninvasive methods were used to the investigated patients with type 2 diabetes mellitus (2-DM) (Group 1), patients with hypertension (Group 2) and patients with both 2-DM and hypertension (Group 3), as well as a normal control group (Group 4) by studying a brachial artery without evidence of atherosclerotic plaque formation. Results　Results showed that endothelium-dependent vasodilation decreased significantly in Group 1 (5.74%±3.32%, P<0.05), Group 2 (4.14%±2.93%, P<0.01), and Group 3 (2.78%±2.08%, P<0.001) as compared to the control (9.45%±3.88%). The nitroglycerin-induced vasodilation (smooth muscle function) in Group 3 was significantly decreased as compared to the control group (14.11%±4.63% vs 23.53%±6.77%, P<0.001), but there were no differences in nitroglycerin-induced vasodilation between Group 1, Group 2 and Group 4. On univariate analysis, a reduced vasodilator response to nitroglycerin was associated with endothelium-dependent vasodilation (r=0.54, P<0.001). Conclusion　Our results indicate that the interaction of 2-DM and hypertension aggravates endothelial dysfunction and further impairs the smooth muscle function.

Objective To develop a quantitative immunohistochemistry assay for duck hepatitis B virus core antigen (DHB-cAg)in duck liver tissue.Methods By comparison with no repair antigen and repair antigen with high pressure,microwave and trypsin,the best solution of antigen retrieval was determined.By optimizing the parameter of image acquisition and de-ducting blank area,mean density of yellow areas was calculated using Image-Pro Plus 6.0 software.Using the assay devel-oped to determine the level of DHBcAg in liver tissue from duck infected by DHBV,anti-DHBV activity of DHBcMAb-TAT PTD conj ugate was examined.Results SABC method with no repair antigen was selected,which was better than other methods.DHBcAg expression in duck liver tissue could be objectively and accurately quantified by setting Image-Pro Plus 6.0 software parameters and calculating mean density of yellow areas.By comparison with the differences between mean densityat baseline of treatment and end of treatment,it was showed that DHBcMAb-TATPTD conjugate treatment dose-de-pendently reduced the levels of DHBcAg in liver tissue,which show that the assay developed could effectively evaluate the anti-DHBV activity of agent.Conclusion The immunohistochemistry assay developed in this study can objectively and accu-rately evaluate the level of DHBcAg in duck liver tissue.

Objective This study investigated high fat diet influence on the changes of vitamin D receptor (VDR) expression and endothelial nitric oxide synthase (eNOS) in apolipoprotein E-deficient(apoE-/-) mice.MethodsApoE-/- mice and C57BLP6J mice were divide into two groups (normal control and high fat diet),high fat diet group were feed high fat feedstuff.Plasma 25-(OH)D levels were determined by competitive protein binding radioimmunity,VDR expression were determined by immunofluorescence and reverse transcription-polymerase chain reaction.The levels of NO and eNOS were determined by nitrate reductase.ResultsCompared with normal control group,high fat diet caused more severe dam-age of atherosclerosis in wild type mice and apoE-/- mice.In apoE-/- mice,the levels of plasma 25-(OH)D were significantly decreased [(26.44±1.28) ng/mL,(22.68±2.07)ng/mL,(17.46±2.22)ng/mL,(15.88±0.97)ng/mL,P＜0.01],the expression of VDR protein and mRNA were significantly increased[VDR :0.244±0.088,0.346±0.132,0.547±0.128,0.768±0.162;VDRmRNA:0.228±0.083,0.375±0.103,0.451±0.117,0.597±0.131,P＜0.01],and the levels of NO and eNOS were significantly increased[NO:(39.74±4.81)μmol/L,(48.1±5.24 )μmol/L,(67.34±6.14 )μmol/L,(86.74±8.05)μmol/L;eNOS:(8.6±0.77 )U/L,(12.28±1.42)U/L,(15.96±0.92)U/L,(18.68±1.15)U/L,P＜0.01].These changes were more significantly in high fat diet group(P＜0.01).ConclusionsThere were abnormalities of plasma 25-(OH)D level,VDR expression and the level of NO and eNOS in apoE-/- mice.These changes were more significantly in high fat diet group.

OBJECTIVETo investigate the roles of Domain I and Domain II of hepatitis C virus (HCV) 5' untranslated region (UTR) in the translation initiation activity of HCV 5'UTR in different host cell lines.

METHODSThe eukaryotic expression plasmid pCMVNCRLuc (pCN1), in which full-length HCV 5'UTR regulates firefly luciferase expression, was modified by deleting Domain I and the downstream single-stranded sequence (43 bp in total) from the UTR (pCNl-d2), Domain I with the downstream single-stranded sequence and Domain II (118 bp in total) from the UTR (pCNl-d3), or the total UTR (pCNl-d5). The modified plasmids were transfected via liposome into different cell lines with pRL-TK plasmid co-transfected as the normalization control. At 36 h after the transfection, the total cellular RNA was harvested for semi-quantitative RT-PCR, and the relative expression activities of luciferase were assayed with a dual luciferase reporter gene assay system. The translation initiation activities of the truncated HCV 5'UTRs in different translation systems were analyzed.

RESULTSDeletion of Domain I and the downstream single-stranded sequence caused no significant changes of the translational activity of HCV 5'UTR in Hela or C6 cells, but decreased the translational activity by 46% in L-02 cells and increased the translational activity by 46% in 293T cells. Deletion of both Domain I and Domain II resulted in decreased translational activity of HCV 5'UTR by 51% in HeLa cells, but increased the translational activity by 40% in L-02 cells, 60% in C6 cells and 135% in 293T cells.

CONCLUSIONSDomain I and Domain II of HCV 5'UTR perform cell type-specific roles in HCV IRES-driven translation initiation.

5' Untranslated Regions ; Genes, Reporter ; HeLa Cells ; Hepacivirus ; genetics ; Humans ; Luciferases ; Plasmids ; Protein Biosynthesis ; genetics ; RNA, Viral ; genetics ; Transfection