Objective To explore the effects of hypericin on inhibition of hepatitis 15 virus (HBV) replication.Methods The concentration gradient hypericin was added to HepG2.2.15 cell culture system and lamivudine was used as control.Enzyme-linked immunosorbent assay (ELISA) and Southern blot were used to examine HBsAg,HBeAg and HBV DNA level in the culture supernatant,respectively.Half inhibitory concentration (IC50) and half effective concentration (EC50) of hypericin were calculated.The effects of hypericin on HBV DNA polymerase were detected by 32p marked deoxy-ribonucleoside triphosphate as the substrate. Independent sample t-test and single factor analysis of variance were used to compare the data between two groups and among multiple groups.Results The inhibition rates of hypericin on HBsAg, HBeAg were enhanced with hypericin concentration increasing and those were higher than lamivudine control group when the concentration was higher than 0.5 μmol/L (t=-0.127,P＜0.05).Southern blot confirmed that hypericin was stronger in inhibition of HBV DNA (EC50=0.2 μmol/L) than lamivudine (t=-0.058,P＜0.05).Hypericin was non-toxic on HepG2.2.15 cells in the range of test with EC50 of 0.2 μmol/L and IC50 of 200 μmol/L.Hypericin did not act on HBV DNA polymerase which was quite different from lamivudine.Conclusions Hypericin can effectively inhibit HBV replication as well as antigen synthesis and is non-toxic on HepG2.2.15 cell.The anti-HBV target of hypericin is different from nucleos(t)ide analogues.

To elucidate the role of reactive carbonyl compounds in the pathogenesis of vascular complication in patients with chronic renal failure or diabetes. Vascular endothelial cells (VECs) were isolated from human umbilical vein and cultured with 3-deoxyglucosone (3-DG) or methylglyoxal(MGO) in vitro. Expression of ICAM-1 and VCAM-1 on the surface of VECs was detected by flow cytometer. The results showed that up-regulation of expression of ICAM-1 and VCAM-1 was observed when either 3-DG or MGO was added into the cultures, which was inhibited by a carbonyl compounds scavanger, aminoguanidine. These data suggested that accumulation of reactive carbonyl compounds in patients with chronic renal failure or diabetes might be involved in the mechanism of arteriosclerosis.

Objective:To investigate the expression of NKG2D in peripheral blood of patients with ovarian benign or malignant tumors, and the expression of the human MHC class Ⅰ chain-related gene A(MICA) on the correspondent tumor tissues. To analyze and discuss the function of NKG2D in anti-ovarian cancer mechanism and its relation with immune escaping of cancer.Methods:Flow cytometry analysis was used to detect NKG2D in the peripheral blood of 42 ovarian carcinoma patients, 23 ovarian tumor patients and 20 health persons. The expressions of MICA in part of the correspondent tissues were examined by means of reverse transcription-polymerase chain reaction (RT-PCR).Results:The expression of NKG2D were (94 23?6 02)%?(98 70?0 98)%?(98 61?1 59)% respectively, compared with the other two groups, the NKG2D expression in malignant group was significantly low; The rate of MICA mRNA expression in ovarian carcinomas was significantly higher than that in benign and normal tissues. No evident relation was found between the expression of MICA mRNA and the clinical factors.Conclusion:The activity of NK cell and the anti-cancer cellular immunity level reduce in patients with ovarian cancer; The decrease of the receptor NKG2D is a reason for the descend of the activity of NK cells; MICA mRNA expression is relative to malignant transformation; the immune-escape of ovarian cancer probably is relative to the down-regulation of NKG2D and the up-regulation of its ligand MICA.

To elucidate the role of reactive carbonyl compounds in the pathogenesis of vascular complication in patients with chronic renal failure or diabetes. Vascular endothelial cells (VECs) were isolated from human umbilical vein and cultured with 3-deoxyglucosone (3-DG) or methylglyoxal(MGO) in vitro. Expression of ICAM-1 and VCAM-1 on the surface of VECs was detected by flow cytometer. The results showed that up-regulation of expression of ICAM-1 and VCAM-1 was observed when either 3-DG or MGO was added into the cultures, which was inhibited by a carbonyl compounds scavanger, aminoguanidine. These data suggested that accumulation of reactive carbonyl compounds in patients with chronic renal failure or diabetes might be involved in the mechanism of arteriosclerosis.

Background Trans-arachidonic acids (TAAs) are specific lipid markers of nitrative stress and the major products of NO2·-mediated ismoerization of aracchidonic acid(AA),and they represent a possible new mechanism of NO2·-induced toxicity in ischemic retinopathy.Our previous study demonstrated that high glucose induce nitrative stress response. Objective This study aimed to evaluate the effect of nitrative stress on retinal capillary in diabetic rats by detecting the TAAs with gas chromatography and mass spectrometry(GC/MS).Methods One hundred 2-week-old clean male SD rats were randomly divided into control group and diabetic group.Diabetic rat models were induced by intraperitoneal injection of streptozoticin of 60 mg/kg,and the successful models were identified as the blood glucose level>16.7 mmol/L.The synthesis method was modified in this study andaraehidonic acid w8$utilized as starting material.14E-AA was obtained as a standard sample via expoxidation and deoxidation in a safe and practical pattern.Serum TAAs and AA in diabetic rats were detected by GC/MS in 2,4,8,12 and 16 weeks after injection of streptozoticin.The ratio of peak area of TAAs to AA with selected ion of 79 was estimated and compared with normal rats. Results Fifty rats had been in Diabetic models were established in 50 rats with the successful rate 100%,and the high blood glucose level in model rats retained throughout the experimental duration.GC/MS results showed that the 14E-AA was obtained with the purity of 97.3%,which provided a standard sample to nitrative stress-related research as a premise. No significant differences were found in serum TAAs/AA values at 2 weeks and 4 weeks between model group and control group (t =-0. 376, t =-0. 642,P>0. 05). However,serum TAAs/AA values in at 8,12 and 16 weeks after streptozoticin injection were considerably elevated in comparison with those of control group (0. 0832 vs 0. 1042,0. 0910 vs 0. 1568, 0. 1100 vs 0. 1895;t=-36.409,t=-166.714,t=-148.212,P<0. 05). Conclusion Elevation of serum TAAs/AA is associated with diabetic duration,implying that damage of nitrative stress to retinal capillary is a parallel procedure to diabetic course.Establishment of GC/MS detection system offers a new evaluating indicator in the research on microvascular ischemic disease and could be implemented in clinical testing.

Medicine, Chinese Traditional ; methods ; Systems Theory

Objective In order to observe the role of genetically modified Schwann cell (SC) with pSVPoMcat in the regeneration of injured spinal cord.Method The cells were implanted into the spinal cord.Ninety SD rats were used to establish a model of hemi- transection of spinal cord at the level of T8,and were divided into three groups,randomly, that is,pSVPoMcat modified SC implantation(Group A), SC implantation(Group B),and without cell implantation as control(Group C).After three months the presence of axonal regeneration of the injured spinal cord was examined by means of horseradish peroxidase(HRP)retrograde labeling technique and stereography.Result The results indicated that HRP labeled cells in Group A and B could be found in the superior region of injured spinal cord and the brain stem such as the red nuclei and oculomotor nuclei. The density of ventral horn neurons of the spinal cord and the number of myelinated axons in 100 μ m of the white matter was A >B >C group.Conclusion In brief,the pSVPoMcat modified SC intraspinal implantation could promote regeneration of the injured spinal cord.

Objective:To establish a method for isolating adipose derived stem cells(ADSCs)from resected human subcutaneous adipose tissues.Methods:ADSCs were isolated,cultured,and expanded from human subcutaneous adipose tissues.Immuno-fluorescent staining of specific molecules.FACS and multi-lineage differentiation induction were used to characterize the obtained ADSCs.Results:ADSCs obtained in this study had the characteristics of stem cells and expressed specific molecules;they also possessed a multi-lineage differentiation potential,which was genetically stable.Conclusion:ADSCs can be isolated from human subcutaneous adipose tissues,which provides a novel and abundant seeding cells for tissue engineering.

Objective To establish a method for determination of trace bromate in flour and the products by ion chromatography.Methods Conductance-ion chromatography and Ionpac~R AS9-HC column were employed and 9.0 mmol/L sodium carbonate was used as the eluent,the flow rate was 1.0 ml/min.BrO_3~-in the samples soaked in the purified water was extracted by ultrasonic microwave,the water-soluble macromolecules were removed by ultra filtration cup in the extract,then the direct injection determination was conducted.Results The lowest detection limit was 0.01 mg/L,the quantitative detection limit was 0.03 mg/kg,the relative standard deviation was 0.6%-7.6%,the recovery rate was 97.8%-103.6%.Conclusion This method is simple,rapid,accurate,sensitive and is applicable to the determination of trace bromate in flour and the products.

Objective To evaluate the plasma level of amino-terminal pro-brain natriuretic peptide(NT-proBNP) before and after treatment in children with dilated cardiomyopathy (DCM) and explore the relationship between NTproBNP and heart functional classes.To evaluate the significance of dynamic change in NT-proBNP level in children with dilated cardiomyopathy.Methods The study comprised 49 children with DCM from Aug.2011 to Sep.2013 in Hunan Children's Hospital.NT-proBNP levels were measured before and after treatment.All the patients were evaluated under the modified Ross Heart Association functional class and echocardiographic determination of left ventricular ejection fraction(LVEF) and fractional shortening(SF).The patients were divided into improvement group(38 cases) and the non-improvement group(11 cases) according to cardiac function after treatment.Results The plasma level of NTproBNP was positively correlated with Ross heart failure classification (r =0.756,P ＜ 0.01),and much better than LVEF and SF.To identify mild heart failure,the cut-off value of 329 ng/L was 93.8％ sensitive and 90.9％ specific.To identify moderate heart failure,the cut-off value of 2 833 ng/L was 95.6％ sensitive and 92.9％ specific.To identify severe heart failure,the cut-off value of 12 125 ng/L was 80.6％ sensitive and 84.8％ specific.There was significant difference in the improvement group before and after treatment(t =2.357,P ＜ 0.05).In non-improvement group there was no significant difference (t =0.132,P =0.778) before and after treatment.Conclusions There is significant correlation between the plasma level of NT-proBNP and heart failure severity.The dynamic changes in NT-proBNP levels before and after treatment can help us to determine the effect of treatment and prognosis.