Objective: To clarify the changes of content and composition of tocopherol in rapeseed oil during the refining process. Methods: The content and composition of tocopherol was determined by HPLC. Results: The content of tocopherol in oil is: crude oil 0.1298%, degummed and neutralized oil 0.1189 %, bleached oil 0.0825%, deodorized oil 0.0686%. Compared to crude oil, the total tocopherol content decreased 47.1% after the whole refining process,and the ?-tocopherol content decreased 33.1%,66.4%,66.7% after degumming and neutralization, bleaching, deodorization, respectively. Conclusion: The content of tocopherol and ?-tocopherol decreases significantly as well as the relative proportion of ?-tocopherol in total tocopherol after refining.These changes must be taken into account during the refining technology optimization, utilization of by-products and evaluation of oxidant stability of rapeseed oil .

ObjectiveTo study the toxic effect of bupivacaine encapsulated in liposomes on spinal cord in rats.MethodsOne hundred and eight SD rats (200-225 g) in which intrathecal (IT) catheter was successfully implanted without complications were randomly divided into 6 groups ( n =18 each):control group (group C) ; liposome group (group L) ; 0.5％ and 1.0％ bupivacaine groups (groups B1 and B2 ) and 0.5％ and 1.0％ bupivacaine encapsulated in liposomes groups (groups LB1 and LB2 ).In groups L,B1,B2,LB1 and LB2,liposome,0.5 ％ bupivacaine,1.0 ％ bupivacaine,0.5 ％ liposomal bypivacaine and 1.0 ％ liposomal bupivacaine 20 μl were injected IT respectively once a day for 7 consecutive days,while in group C nothing was injected IT.Pain threshold was measured by mechanical stimulation of the plantar surface of hindpaw.Motor function of the hindlimbs was also assessed.The animals were sacrificed at 8 day after IT injection.The lumbar segment of the spinal cord was removed for microscopic examination,detection of neuronal apoptosis (by flow cytometry) and Fos protein expression (by immuno-histochemistry).Results1.0％ bupivacaine IT significantly increased the percentage of apoptotic neurons in group B2 as compared with control group.0.5％ and 1.0％ bupivacaine IT significantly increased the number Fos protein positive cells in group B1 and B2 as compared with group C.1.0％ bupivacaine IT induced severe histologic damage including shrinkage of nucleus and vacuole formation in mitochondria.Encapsulation of bupivacaine in liposomes significantly attenuated bupivacaine-induced increase in apoptosis and Fos protein expression and histologic damage in group LB2 as compared with group B2.ConclusionThe encapsulation in liposomes can decrease the neurotoxicity of 1.0 ％ bupivacaine administered IT in rats.

Objective To study the reliability and validity of the clinical neurologic deficit scale in evaluating stroke patients. Methods A total of 222 inpatients with acute stroke onset were included in the study. They were assessed when admitted, at the 14th and 90th day of hospitalization by different physicians using the clinical neurologic deficit scale. Intrarater and interrater reliability were determined using Kappa correlation. The split-half rehability and internal consistency were evaluated using Cranbach's a coefficient. Concurrent validity and the predictive validity were determined by spearman rank correlation coefficients. Construct validity was assessed by the factor analysis and the construct validity of the scale was measured according to the classifications of the Oxfordshire Community Stroke Project ischemic stroke subtypes in the patients with cerebral infarction. Results The scores of intrarater reliability in all items were higher than 0.6, the score of interrater reliability in the item "walking" was 0.542, the split-half reliability and the internal consistency were good as demonstrated by the score of 0.911 and 0.886 respectively, and assessment of reliability of different methods showed that "strength in upper limb" and "strength in hand", were poor as shown by the score of 0.393 and 0.386 respectively. The scale is highly correlated with the NIHSS ( both P=0.000) in both total and subtypes of stroke patients according to the classifications of the Oxfordshire Community Stroke Project by concurrent validity analysis. There was a high correlation between the scores of the scale and Barthel Index and the modified Rankin scales at the 90th day of hospitalization (both P=0.000). Conclusions The clinical neurologic deficit scale has a good internal consistency. There is concurrent validity between the scale and the NIHSS and could predict stroke outcome. Factor analysis of the scale displays the best construct validity in the patients with partial anterior circulation infarction, and could be used to evaluate the focus of vertebrobasilar artery despite its insensitivity.

Activities of N-acetyl-?-glucosaminidase (NAG) in sera of normal subjects and patients with hepatocellular carcinoma (HCC) and other diseases were microquantitatively determined by spectrophotometry. The results showed that after the reaction for 4 h, activity of serum NAG in patients with HCC (397.10 ? 174.97 nmol/ml) was obviously higher than that in normal subjects (280.00 ? 63.83 nmol/ml). The elevation of serum NAG in HCC patients had no relationship with their serum a-fetoprotein. However, the increase of serum NAG activity in some patients with benign liver diseases was always accompanied by the abnormality in their liver functions. It is suggested that serum NAG has some referable values in the diagnosis of HCC.

Objective To investigate the relationship between the polymorphism of transforming growth factor-β1 (TGF-β1 ) gene and risk of chronic hepatitis B virus(HBV)infection progressing to hepatocellular carcinoma(HCC).Methods 120 patients with chronic HBV infection(case group)and 100 age-and sex-matched healthy individuals(healthy control group)were randomly enrolled to this study.The polymerase chain reaction-restriction fragment length polymorphism technique was adopted to detect the single nucleotide polymorphism of TGF-β1 gene(T29C),and made the comparative analysis combined with TGF-β1 mRNA level. Results The risk of HCC occurrence in the patients carrying genotype CC was decreased than that in the patients carrying geno-type TT (OR=0.317,95%CI =0.110-0.913,P =0.033;OR=284,95%CI =0.093 -0.866,P =0.027),the risk of HCC in pa-tients carrying allele C was significantly decreased compared with that in the patients carrying allele T(OR =0.570,95%CI =0.341 -0.953,P =0.032;OR=0.548,95%CI =0.320-0.936,P =0.028).In the HCC group,the patients carrying genotype CC had the lower lever of TGF-β1 mRNA.Conclusion TGF-β1 gene polymorphism(T29C)may be related to the risk of chronic HBV infection progressing to HCC.

Objective To compare the effect of perioperative intravenous injection of Analgecine on the analgesic efficacy and complications of patient-controlled intravenous analgesia ( PCIA ) of different doses of fentsnyl in postoperative lumhotomy patients.Methods 200 patients underweat hmbotomy in general anesthesia were randomly divided into four groups with fifty cases each.Fentanyl 1.0mg in group A,fentanyl 0.5mg in group B,fentanyl 1.0mg in group C,fentanyl 0.5mg in group D.The drugs in each group were diluted to 100ml and infused by pumps.Besides,the patients in group C and D were injected with analgecine 3.6u and 7.2u at the night before the operation,preoperation and postoperation respectively.The visual analog scale(VAS),times of PCA and incidence of side effects were recorded during the period of postoperative 24 hours.Results The VAS of group B at 2h after operation was (5.2 ± 1.9 ) points,which was significandy higher than that of group A,C and D( P ＜ 0.05 ),VAS became similar 4h later( P ＞0.05).The demanding times for supplemental bolus in group B were also significantly higher than that of A,C and D( P ＜ 0.05 ).The incidence of nausea,vomiting,itching,somnolence in group B and D were significantly less than those in group A and C( P ＜0.05 ).No respiratory depression or abnormal bleeding occurred in the four groups.Conclusion Perioperative intravenous injection of analgecine had a better effect on PCIA of fentanyl and could reduce fentanyl requirement and its side effects in lumbotomy patients.

Objective To investigate serum NK cell levels in HBeAg positive early pregnancy women with immune activation.Methods Fifty four HBeAg positive pregnant women admitted in Taizhou People's Hospital from September 2010 to April 2013 were enrolled in the study.Among them,the serum HBV DNA load decreased ≥2 log at 12 weeks after pregnancy in 24 cases (immune activation group) and HBV DNA did not decrease in 30 cases (immune tolerance group).The serum level of alanine aminotransferase (ALT),total bilirubin,HBeAg,HBV DNA load and NK cells were measured.Results At week 12 of gestation,the mean ALT levels and ALT abnormality rate in immune active group were higher than those in immune tolerance group [(146.7 ±93.1) vs.(44.1 ± 14.7) U/L,t =2.95,P＜0.05,50.0％ vs.6.7％,x2 =4.97,P ＜0.05].There was no significant difference of HBeAg level between two groups before pregnancy,while HBeAg level in immune activation group was lower than that in immune tolerance group at week 12 week of gestation [(291.8 ± 170.5) vs.(443.7 ± 289.9) S/CO,t =2.81,P ＜0.05].The percentage and absolute number of NK cells in immune activation group were higher than those in immune tolerance group [(26.7 ±9.1)％ vs.(17.1 ±7.8)％,t =2.52,P ＜0.05 and (370.9 ±136.4)/μl vs.(213.2 ±97.8)/μl,t =2.38,P ＜0.05,respectively].Conclusions In HBeAg positive early pregnant women with immune activation,the inhibition of HBV DNA might be associated with the activation of NK cells.

Objective To investigate the therapeutic effect and the detailed mechanisms of intraarterially delivery of bone marrow mesenchymal stem cells ( BMSCs) for treatment of middle cerebral artery occlusion (MCAO) in rats.Methods BMSCs were isolated,purified and amplified with the adherence culture method.BMSCs were labeled with 5-bromo-2-deoxyuridine ( BrdU ) (10 μmol/L) for 48 h before transplation.Surface antigens of CD90,CD29,CD106,CD34,CD45,CD11b were identified by flow cytometry.The MCAO model was established with suture emboli method.In this study,3×106 BMSCs were injected into rats with MCAO through intraarterial route at day 7 after stroke.The effects on functional and physical recovery were assessed with the behavioral tests (mNSS test and adhesive test) and body weight.Bielshowsky-Luxol Fast Blue double staining was used to demonstrate the reconstruction of axon and myelin.The Brdu-labeled BMSCs in vitro and in vivo were detected with direct immunofluorescent staining.The expression of neuron specific enolase ( NSE),neurite outgrowth inhibitor-A ( Nogo-A),synaptophysin (SYN),ki-67 nuclear antigen (Ki-67),glial fibrillary acid protein( GFAP),vascular endothelial growth factor ( VEGF) in brain were analyzed with immunohistochemical staining.Results Flow cytometry indicated that the positive rates of high expression of CD90,CD29,CD106 in BMSCs were respectively 91.70%,88.40% and 52.20%.Meanwhile,the positive rates of low expression of CD34,CD45,CD11b in BMSCs were 2.70%,5.65% and 7.82%,respectively.There was a significant difference in behavioral tests ( mNSS test and adhesive test) between BMSCs group and PBS group at day 21,28,35 after MCAO (mNSS:4.89 ±1.36,7.00 ±1.67,3.78 ±1.30 and 6.33 ±1.21,2.44 ±1.13,5.67 ± 1.51;t =2.69,3.83,4.75;adhesive test:54.00 ± 10.48,68.17 ± 11.09,36.89 ±9.80 and 59.33 ± 12.40,23.44 ± 9.04,46.50 ±9.38;t =2.51,3.92,4.77;P ＜0.05).Meanwhile,a significant difference in body weight was discovered between them at day 28,35 after MCAO.In BMSCs group,the area of corpus callosum in the ipsilateral hemisphere was significantly enlarged,the positive number of Brdu,SYN,Ki-67,GFAP,VEGF in brain was significantly increased,the expression of Nogo-A in brain was significantly decreased,nevertheless,the number of NSE-positive cells in brain and the infarct volume were not significant different from PBS group at day 35 after MCAO.Conclusions These results suggest that intra-arterial transplantation of BMSCs is an efficient treatment protocol for stroke.Treatment with BMSCs increases endogenous cells proliferation,angiogenesis,synaptogenesis,enhances axonal regeneration and the protective function of astrocytes,all of which may contribute to neurological functional recovery.

Objective To investigate the correlation between the glial fibrillary acidic protein(GFAP),neuron-specific enolase(NSE),synaptoghysin (SYN),neurite outgrowth inhibitor-A(Nogo-A)expression and neurological outcome in tissue surrounding the infarct during the recovery after cerebral ischemia-reperfusion injury in rats.Methods A 2-hour middle cerebral artery occlusion(MCAO)and reperfusion model in rats was induced by the intraluminal suture method.The modified neurological severity score(mNSS)was performed at day 28,35,42,and 49.Immunohistochemistry was used to detect the expressions of GFAP,NSE,SYN,and Noga-A in tissue surrounding the infarct.Results The mNSS score decreased gradually over time after cerdnal ischemia-reperfusion injury in rats.Except day 35(5.11±0.737)vs.day 42 (4.54±0.519),and day 42 vs.day 49(4.29±0.488),there were significant differences at all other time points(all P<0.05).The numbers of GFAP positive cells deergased gradually form day 28 to day 49,in which,the numbers of GFAP positive cells at day 42(51.00±13.59)vs.day 49(44.38±11.94) were significantly less than those at day 28(69.00±15.10)(P<0.05).There were no significant differences in the numbers of NSE positive cells at all time points,but their integrated optical density(IOD)increased gradually.There were significant differences between day 28(6 218.57±1 864.25)and day 42(9 414.00±2 491.12) or day 49(12 522.50±3 106.99),and between day 35(7 343.40±1 533.35)and day 49(all P< 0.05).There were no significant differences at all other time points.The SYN express (IOD)increased gradually.and it was significantly lower at day 49(66 503.00±12 834.61)than that at day 28(43 905.14±13 208.59)(P<0.05).The numbers of Nogo-A positive cells decreased gradually,and they were significantly less at day 49(42.13±14.45) than those at day 28(59.57±15.25)(P<0.05).The GFAP expression was positively correlated with the mNSS scores(r=0.993,P=0.007).The NSE(r=-0.954,P=0.044)and SYN(r=-0.992,P=0.008) expression was negatively correlated with the mNSS scores.Conclusion The neurological outcome was associated with the downregulation of GFAP expression and the upregalation of NSE and SYN expression during the recovery after cerebral ischemia-reperfusion injury in rats.