We hypothesized that the concentrated urea and NaCl solutions may opened the blood-brain barrier to the horseradish peroxidase and trypan blue-albumin complex by shrinking barrier cells and opening up spaces between them. The experiments were carried out on 30 healthy, adult rats. Two experimental groups were used. First, intracarotid perfusion of anesthetized rats were prepared by exposing and catheterizing the left common carotid artery. A test solution of 3.4 M, 3.0 Osm urea and 0.87 M, 1.6 Osm NaCl, was perfused manually for 30 sec. in a cranial direction so as to expel the blood from the pial arterioles of the exposed brain. The pressure, which was not measured, varied between what was required which to expel blood from both arterioles and venules. Five milliliters of the test solution usually were used. Horseradish peroxidase and trypan blue was injected intravenously or through the carotid artery after perfusion. Threshold of barrier damage due to the intracarotid substance was defined as the lowest osmotality which produced obvious blue staining of the brain both on surface observation and coronal section. The effect of a substance was defined as reversible if a threshold concentration did not produce blue staining when the dye was injected 30 min following perfusion. Second, we applied a concentrated solution of 3.0 Osm urea and 1.6 Osm NaCl to the pia-arachnoid of the cerebral cortex, to study the barrier to the intravascular horseradish peroxidase and trypan blue-albumin complex. Hyperosmotic solution of 3.0 Osm urea and 1.6 Osm NaCl, either infused into one internal corotid artery or applied topically to the pia-arachnoid surface of the brain of rats, results in the opening of endo thelial tight junction through which horseradish peroxidase passes from blood to the basal menbrane and astrocytes and neurons. The evidence for this opening of the blood-brain barrier to protein is the entry of peroxidase into the neurons. It was postulated that sufficiently high concentrations of electrolytes or relatively lipid-insoluble non-electrolytes such as urea, osmotically pulled water from the cerebral endothelial cells resulting in their shrinkage. The shrinkage, in turn, was believed to open the tight junction between continuous endothelial cells so that the dyeprotein complex could pass through the junction from blood to neurons. The present study shows that these tight junctions, the sites of the barrier to neuron movement of protein, are indeed opened by the osmotic action of urea or NaCl.

OBJECTIVE:To study the pharmacokinetics of oral single dose adefovir dipivoxil(ADV) tablet in Chinese healthy volunteers.METHODS:The study was a randomized,open,three-way crossover study.Twelve healthy volunteers were randomly assigned to receive single oral dose of 5 mg(Group A),10mg(Group B) or 30 mg(Group C) ADV tablets in three weeks.The plasma concentrations of ADV were determined by LC/MS/MS method.The pharmacokinetic parameters were computed.RESULTS:The main pharmacokinetic parameters in Group A,B and C were as follows:Cmax were(11.4? 3.7),(25.4? 8.2) and(76.3? 23.0) ng? mL-1;tmax were(1.69? 1.41),(0.90? 0.56) and(0.94? 0.50) h;AUC0～ t were(102.7? 51.7),(235.0? 82.3) and(715.4? 267.6) ng? h.mL-1;AUC0～ ∞ were(168.7? 30.7),(266.2? 83.7) and(741.5? 273.9) ng? h? mL-1,respectively.CONCLUSION:ADV tablets had a rapid absorption in healthy volunteers and the Cmax and AUC of adefovir tablets were directly correlated to doses.It is safe for healthy volunteers to take ADV tablets at a dose of 5～30mg.

[Objective]To evaluate the two-dimensional shear wave elastography(2D SWE)in predicting the prognosis of the patients with acute-on-chronic hepatitis B liver failure(ACLF-HBV).[Methods]312 patients diagnosed with ACLF-HBV at the De-partment of Infectious Diseases in the Third Affiliated Hospital of Sun Yat-sen University from October 1st 2013 to December 31th 2015 were recruited. The baseline data of all patients,including 2D SWE,laboratory data,ultrasonographic data,Model for End-stage Liver Disease score(MELD),Child-Turcotte-Pugh score(CTP),and King′s College Hospital score(KCH),were collected when enrolled. All patients were followed up for at least 90 days and their clinical outcomes(recovering or worsening)were recorded. 2D SWE values of all patients were dynamically collected every 2~4 weeks during the follow-up until death,liver transplantation, or discharging from hospital.[Results]The worsening patients showed higher 2D SWE values than the recovering ones[(44.0 ± 7.5) kPa vs(36.8 ± 10.3)kPa,t=-6.5,P=0.000],2D SWE value less than 39.2 kPa could be a prediction of the patient′s recovery in 90 days.The predictive value of 2D SWE(AUROC=0.73)was higher than that of KCH(AUROC:0.65,z=2.1,P=0.033). Among all the dynamically measured patients,2D SWE rose from(42.1 ± 8.0)kPa to(47.5 ± 9.3)kPa in the worsening group during the 90-day follow-up,while nearly stable in the recovery group.[Conclusion]Noninvasive liver stiffness measurement by 2D SWE is a use-ful method in evaluating the prognosis of ACLF-HBV patients. Taking a cutoff of 39.2 kPa,patients with higher 2D SWE values could have worse clinical outcomes ,otherwise better. It seems that dynamically measuring 2D SWE values could also be helpful ,but more research in the future should be needed.

on,size and the extent structures of atrial septal defect on line.

Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.

Objective To evaluate the short-term efficacy of lamivudine versus entecavir for patients with HBeAg-negative acute-on-chronic liver failure (ACLF) with different pretreatment liver failure degrees.MethodsA total of patients with HBeAg-negative ACLF were enrolled into this retrospective study.Seventy-two cases were treated with lamivudine 100 mg daily,while 93 cases were treated with entecavir 0.5 mg daily.Biochemical items,model for end-stage liver disease (MELD)score,hepatitis B virus (HBV) DNA level and mortality were observed.The efficacies of the two drugs were analyzed in patients with different degrees of liver failure.The comparison of rates was done using chi-square test and the measurement data were compared by t test.ResultsAmong the patients with pretreatment MELD scores above 30,the post-treatment HBV DNA levels in lamivudine group and entecavir group were (3.6 ± 1.1) lg copy/mL and (3.7 ± 1.4) lg copy/mL,respectively (t=0.181,P=0.859) and the mortalities were 92.0％ and 91.8％,respectively (χ2 =0.002,P=0.680).For the patients with pretreatment MELD scores from 23 to 30,the post-treatment HBV DNA levels in two groups were (3.2± 1.1) lg copy/mL and (3.2±2.3) lg copy/mL,respectively (t=0.760,P=0.455) and the mortalities were 42.9％,54.1％,respectively (χ2 =0.799,P=0.455).In patients with pretreatment MELD scores below 23,the post-treatment HBV DNA levels in two groups were (3.1±1.0) lg copy/mL and (2.8±1.5) lg copy/mL,respectively (t=-0.740,P=0.464) and the mortalities were 3/19 and 6.3％,respectively (χ2=1.227,P=0.455).In lamivudine group,the mortalities were significantly different among patients with three different ranges of pretreatment MELD scores (χ2 =26.967,P =0.000).The similar differences were also found in entecavir group (χ2 =41.260,P=0.000).ConclusionsAmong treatment na?ve patients with HBeAg-negative ACLF,the short-term efficacy of lamivudine versus entecavir is equal if the degree of pretreatment liver failure is similar.Meanwhile,the degrees of pretreatment liver failure significantly affects the outcome of the treatment.

OBJECTIVE:To study the alleviation effect of nervonic acid on movement disorder of model mice with Parkinson's disease(PD). METHODS:Mice were randomly divided into blank control group(normal suline),model group(normal saline), Levodopa and benserazide hydrochloride tablet group (positive control,calculated by L-dopamine 120 mg/kg),nervonic acid low-dose,medium-dose,high-dose groups(20.0,40.0,80.0 mg/kg),10 in each group. Except for blank control group,mice in other groups were inducced for PD models. After modeling,mice were intragastrically given relevant medicines,once a day,for 14 d. After the last administration,behavioral changes of mice in each group were observed. HPLC was conducted to detect dopa-mine(DA)and its metabolites dihydroxybenzoic acid(DOPAC),homovanillic acid(HVA)concentrations in the striatum of mice. RESULTS:Compared with blank control group,climbing time was extended in model group,drum time was shortened,spontane-ous movement times was decreased,and DA,DOPAC,HVA contents in the striatum were reduced (P<0.05). Compared with model group,climbing time was shortened in Levodopa and benserazide hydrochlo ride tablet group,nervonic acid dose groups, drum time was extended,and DA,DOPAC,HVA contents in the striatum were increased(P<0.05);and spontaneous movement times was increased in Levodopa and benserazide hydrochloride tablet group,and nervonic acid high-dose group(P<0.05). CON-CLUSIONS:Nervonic acid can effectively improve symptoms of movement dysfunction of model mice with PD. The mechanism may associate with increasing DA content in the striatum.

Antimony ; urine ; Flow Injection Analysis ; Spectrometry, Fluorescence ; methods ; Spectrophotometry, Atomic ; methods

ObjectiveTo investigate the relationship of lymphotoxin β receptor (LTβR) and classical nuclear factor-κB (NF-κB) activation pathway in the pathogenesis and progress of cystitis and bladder cancer.MethodsThe LTβR and P65 mRNA expression were detected by Real-time quantitative PCR in 108 cases of fresh bladder tissue specimens (75 cases of bladder cancer,10 cases of inflammation and 23 normal bladder mucosa cases grouped by the tissue classification ),and protein expression were analyzed by immunohistochemistry assay in 118 cases of paraffin-embedded biopsy specimens (73 cases of bladder cancer,30 cases of cysitis and 15 normal bladder mucosa cases).The correlation analysis between the expressions of LTβR and P65 with clinical pathological data was then performed.Differences between LTβR and P65 mRNA and protein expression level were compared in different groups of bladder tissues using Kruskal-Wallis H test and the Chi-square test.Results( 1 )The mRNA expressions of LTβR and NF-κB/P65were higher in bladder cancer than those in normal group ( LTβR:29.4 ( 14.2 - 46.7 ) × 10 - 3/1.2 ( 0.3 -7.0) ×10-3,Z=-5.508; P65:9.7 (2.7 -21.1) ×10-3/1.0(0.8 ～1.8) ×10-3,Z=-5.030,P＜0.05 ).There were significantly differences between bladder cancer with different histological grades ( LTβR:18.2(2.1-31.3) × 10-3/ 28.4(16.6-36.2) × 10-3/47.9(34.3 -70.5) ×10-3,x2K-W=20.378;P65:4.9(1.3 - 12.0) × 10-3/7.4(3.0-21.9) × 10-3/17.0(10.0 ～28.3)× 10-3 ,x2K-W2 =15.219,P all ＜0.05) and lymph node metastasis (LTβR:27.2(9.7-40.1) ×10-3/39.4(26.7 -52.6) ×10-3,Z=-2.552; P65:7.4(2.3-15.6) ×10-3/13.4(6.7-23.3) ×10-3,Z=-2.026,P＜0.05).(2)The positive rates of LTβR and phosphorylated P65 ( p-P65 ) protein in cancer were higber than those of normal group (LTβR:69.8％/13.3％,x2 =16.600 ; p-P65:56.2％/6.7％,x2 =12.220,P ＜ 0.05 ).Upregulation of LTβR and p-P65 were associated with the histological grade (LTβR:56.3％/70.0％/90.4％,x2 =7.055; p-P65:40.6％ /60.0％/76.2％,x2 =6.679,P ＜0.05) and with lymph node metastasis (LTβR:58.3％/92.0％,x2 =8.849; p-P65:52.1％/64.0％,x2 =5.088,P ＜0.05).(3)There was a positive correlation between LTβR and P65 expression ( mRNA:r =0.654,P ＜ 0.05,protein:r =0.399,P ＜ 0.05 )in the bladder cancer and cystitis (r =0.521,P＜0.05).ConclusionsThe activation of LTβR and P65 was associated with progression and metastasis of bladder cancer.The activation of classical NF-κB pathway by LTβR may be achieved by P65.

End-stage liver disease refers to the advanced stage of liver disease caused by various chronic liver damage. Orthotopic liver transplantation is the most important final treatment option, but liver transplantation is still limited by many factors at present. Stem cell transplantation therapy has attracted widespread attention as a potential treatment for end-stage liver disease. This article reviews the research progress of mesenchymal stem cell therapy in end-stage liver disease.