Objective To investigate the clinical value of the combined detection of manganese superoxide dismutase (Mn-SOD)genetic polymorphism and lipoprotein phospholipase A2 (Lp-PLA2)in the early diagnosis of coronary heart disease (CHD).Methods 92 cases of coronary heart disease patients and 78 cases of healthy control group were selected.The activ-ity of Lp-PLA2,the activity of Mn-SOD and genotype of Mn-SOD 9 Ala/Val genetic polymorphism were detected in the ser-um of each group via the use of colorimetry,continuous monitoring technique and gene-sequencing method respectively and then the correlation of serum Mn-SOD,Lp-PLA2 and Mn-SOD genetic polymorphism with CHD were analyzed.Results The Lp-PLA2 activity in serum of CHD groups with Mn-SOD 9 VV genotype was statistically significantly higher than that of patients with the AV+AA genotype (P<0.01).The serum Mn-SOD activity in patients with VV genotype was signifi-cantly lower than that of those with AV+AA genotype (P<0.01).Conclusion Combined detection of Mn-SOD 9 Ala/Val genetic polymorphism and Lp-PLA2 activityin the serum can provide an important foundation for the diagnosis and predic-tion of coronary heart disease.

Objective To investigate the relationship between the activity of superoxide dismutase (T-SOD)as well as its manganese superoxide dismutase isozyme (Mn-SOD),concentration levels of hypersensitive c-reactive protein(hs-CRP)and coronary heart disease.Methods The levels of serum T-SOD,Mn-SOD and hs-CRP were measured in 81 patients with coro-nary heart disease and 60 healthy controls,respectively.T-SOD was measured by colorimetricmethod and hs-CRP was meas-ured by latex enhanced immune turbidimetric assay.Results Compered with the control group,activity of T-SOD,Mn-SOD in CHD group were significantly decreased (t=9.41,6.34,all P<0.01).However,hs-CRP in CHD group were significantly increased to those in controls (t=3.09,P<0.05).The activity of T-SOD,Mn-SOD were negatively correlated with hs-CRP (P<0.01).Conclusion The variation of T-SOD,Mn-SOD activity and hs-CRP content were closely related to the occur-rence and development of CHD,they could be the impor tant indicators for riskfactors assessment of CHD.Moreover,con-joint analysis the correlation of T-SOD,Mn-SOD and hs-CRP has certain guiding significance for the clinical treatment and prognosis of coronary heart disease.

Objective To investigate the relationship between serum homocysteine (HCY)levels and oxidative stress in pa-tients with coronary heart disease (CHD).Methods According to the diagnostic criteria of CHD in 2010 (WS 319-2010)is-sued by the Ministry of Health,79 patients with CHD in Xi’an Central Hospital from June 2014 to December 2014 were se-lected as the experimental group,and 5 5 cases of healthy physical examination for the same period were selected as the nor-mal control group.The levels of oxidative stress marker malondialdehyde (MDA)and the activity of antioxidant enzyme su-peroxide dismutase (SOD)in serum were detected to analyze the state of oxidative stress.The levels of serum HCY were de-tected,and the correlation between HCY levels and MDA levels and SOD activity was analyzed.Results The levels of MDA in the CHD group were significantly higher than that in the control group,while SOD activity was significantly lower than that in the control group,the differences were statistically significant (t=3.112,2.684,all P<0.05).The levels of HCY in the CHD group were significantly higher than that in the control group,the difference was statistically significant (t=3.268,P<0.05).In the CHD group,the levels of HCY were positively correlated with the levels of MDA (r=0.236,P<0.05),and negatively correlated with SOD activity (r=-0.221,P<0.05).Conclusion Serum HCY levels were increased and oxidative stress reaction was enhanced in patients with CHD.Oxidative stress was associated with elevated serum HCY levels.Serum HCY levels and oxidative stress played an important role in the occurrence and development of CHD.

Object To study the augmentation of the T activated killer (T AK) cell proliferating and tumor killing activities of the polysaccharide fraction from Rhodiola kirilowii (Regel) Regel, Lycium barbarum L., Hedysarum polybotrys Hand. Mazz., Glehnia littoralis F. Schmidt ex Miq., Rehmannia glutinosa (Gaert.) Libosch. ex Fisch. et Mey. and Aloe barbadensis Mill. in vitro. Methods The T AK cells were induced by anti CD 3 antibody (CD 3McAb) and rIL 2 from human peripheral blood mononuclear cells (PBMC). The effects of the above six plant polysaccharides (1～100 ?g/mL) on the proliferation of T AK cell, the cytotoxicity to Raji cells and L 1210 cells, and the IL 2 receptor (IL 2R) expression in T AK cells were determined. Results The six polysaccharides alone had no obvious effect on the proliferation of T AK cells. In the presence of rIL 2 and CD 3McAb, they could reinforce the proliferation of T AK cells and its tumor killing activities against Raji cells and L 1210 cells at a different extent, and a 21%～68% increase of IL 2R expression in T AK cells was observed. Conclusion The plant polysaccharides significantly enhance the proliferation and the tumoricidal activities of T AK cells and the enhancing actions related to the increase of IL 2R expression.

Objective To investigate the relationship between manganese superoxide dismutase (Mn-SOD)9 Ala/Val genetic polymorphisms and the levels of blood lipid and homocysteine (HCY).Methods The genotypes of Mn-SOD 9 Ala/Val ge-netic polymorphisms were identified by sequencing method,the serum activities of T-SOD and Mn-SOD were detected by colorimetric method,the serum level of HCY was detected by enzymatic method,and the serum levels of cholesterol (TC), triglyceride (TG),high density lipoprotein cholesterol (HDL-C)and low density lipoprotein cholesterol (LDL-C)were de-tected by end-point method in 137 patients with coronary heart disease (CHD)and 85 controls.Results Compared with the control group,the VV genotype and V allele of Mn-SOD 9 Ala/Val genetic polymorphisms in the CHD group were higher, while the serum activities of T-SOD and Mn-SOD in the CHD group was significantly lower.The serum activities of T-SOD and Mn-SOD of the Mn-SOD 9 VV genotype was significantly lower than the Mn-SOD 9 AA genotype.Compared with the Mn-SOD 9 AA genotype,the serum levels of TC,TG,LDL-C and HCY of the Mn-SOD 9 VV genotype were significantly higher,while the serum level of HDL-C was significantly lower.The serum activity of Mn-SOD was negativelycorrelated with the serum levels of TC,TG,LDL-C and HCY and positively correlated with the serum level of HDL-C.Conclusion The antioxidative ability in patients with CHD was decreased.Mn-SOD 9 Ala/Val genetic polymorphisms led to lipid metab-olism disorders by affecting the Mn-SOD activity,promoting the development of CHD.HCY resulted in increased oxidative substances by self-oxidation and inhibition of the Mn-SOD activity,increasing the risk of CHD.

Background and purpose:Bladder cancer radical surgery is dififcult with many perioperative complications, and the learning curve is long. To introduce the clinical application of complete retrograde radical cystectomy and consequent abdominal cavity reconstruction in male bladder cancer, and to provide a standardized surgical procedure with minimum perioperative complications and short learning curve. Methods:From Apr. 2012 to Apr. 2013, 110 cases of male patients with bladder cancer received this procedure in our department, with a median age of 64 (35-83) years;Preoperative characters, surgical parameters, perioperative complications, pathology, long-term complications and short-term prognosis were collected and analyzed. Results:The median number of lymph nodes resected in operation was 12 (8-16);Neurovascular bundles were reserved bilaterally in 65 cases, and unilaterally in 31 cases;The complete procedure including urinary diversion took 4.4 (2.2-6.0) hours, with a median time of opened abdominal cavity of 43.0 (5.0-75.0) minutes;The median blood loss was 140.0 (50.0-600.0) mL, and 4 patients needed transfusion; Median time of abdominal and pelvic drainage was 10.0 (6.0-15.0) days, the median gastrointestinal recovery time was 2.5 (1.0-12.0) days, and the median postoperative hospital stay was 17.0 (10.0-39.0) days;Grade 2 Clavien-Dindo classiifcation (CDC) of surgical complications that required medical intervention were found in 19 cases, CDC grade 3 or above were found in 8 cases;Mild to moderate postoperative ileus happened in 5 cases, all recovered in median 2 (1-4) weeks with supportive treatments;There were no perioperative deaths. All samples were sent to pathological analyses. After a median follow-up of 9 (3-15) months, no complications of or above CDC grade 3 happened, and there were no recurrence. Conclusion:Complete retrograde radical cystectomy in male bladder cancer provided clear anatomical approach, reliable neurovascular bundle preservation, less blood loss, limited abdominal organs disturbance and better surgical exposure; With respect to tumor control, more peritoneal was retained for subsequent abdominal cavity reconstruction. The introduced procedure effectively speeded up gastrointestinal recovery, reduced postoperative complications, especially the incidence of ileus and its severity, and shortened hospital stay. The learning curve of this procedure for urologists was short, and further investigation was warranted.

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.

Objective To study the effect of the recombinant Lentivirus containing calcitonin gene related peptide (CGRP) gene on cells biological activity and differentiation of rat bone marrow stem cells(MSCs).Methods Rat MSCs were isolated and cultured by granulocytes adherent.MSCs were transfected with Lenti-EGFP CGRP(MSCsCGRP+/+ group),While MSCs were transfected with Lenti-EGFP as control group.Cell transfection rate was detected by flow cytometry,protein secretion in the above-mentioned MSCsCGRP+ + supernatant was detected using ELISA method.Cells surface markers weare detected by flow cytometry and immunohistochemistry.Trypan blue was used to examin the survive rate,β galactosidase staining was used to examin aging of MSCs transfection,and MTT was used to examine cell vitality.Results At first day after transfecting with Lenti-EGFP-CGRP,fluorescence was not observed by fluorescence microscope,but a small amount of CGRP protein was detected by ELISA in MSCsCGRP+/+ group,at 3 days and 4 days after transfecting with MSCs,strong fluorescence was observed by fluorescence microscope (the cell transfection rates were 77.87％ and 79.58％).The CGRP expression was significantly higher in MSCsCGRP+ + group than in control group [(19.53±0.50) pg/ml vs.(3.12±0.00) pg/ml,t=48.964,P＜0.01].At three days after transfection with MSCs,CD29 and CD90 expression were significantly higher,as compared with control group,CD31 expression was increased in MSCsCGRP+ /+ group.Seven days after transfection with MSCs,CD31 expression was significantly increased in MSCsCGRP+ + group,vWF expression was significantly increased in MSCsCGRP+ + group after MSCs were transfected with LentiEGFP CGRP for 14 days,but a SMA expression was decreased in MSCsCGRP+ +group.At 3 days and 7 days after transfection with Lenti-EGFP-CGRP,the proliferation,survive and aging showed no difference in MSCsCGRP+/+group and in control group (the proliferation of cell:t=0.253,0.290the survive of cell t=-0.307,0.690,all P＞0.05).At 14 days after transfection with Lenti-EGFP-CGRP,aging of cell were decreased in MSCsCGRP+ + group as compared with control group (t=2.446,P＜ 0.05).Conclusions After MSCs are transfected with Lenti EGFP-CGRP,biological characteristics of MSCs has no significant effects,there is still proliferation and differentiation activity.Cell secretion of CGRP can promote the endothelial cell differentiation,and inhibit the differentiation to smooth muscle cells.The CGRP modification of MSCs may play a role in the regulation of angiogenesis.

The free membrane of Eudragit L100/S100 which is pH-sensitive, colon-specific was prepared by plane casting films. The film humidity, species and amount of plasticizers, the ratio of membrane material was investigated. The rate of membrane permeability and mechanical properties were used as indicators of orthogonal experiment, and its related properties were studied. The results show that the mechanical properties of the membrane and phragmoid capacity are the best when 30% TEC was used as plasticizer; the ratio of membrane material have little effect on the rate of membrane permeability and mechanical properties. By adjusting the species and amount of plasticizers, the ratio of Eudragit L100/S100, the free membrane which is colon-specific can be obtained.

BACKGROUND: That the nerve growth factor(NGF) is capable of treating peripheral nerve injury has been broadly acknowledged. But it is not sure whether it is able to pass through blood brain barrier(BBB) to act on central nerve system. In this study, the NGF encapsulated in liposome was compared with NGF alone in their abilities of passing through BBB.OBJECTIVE: To compare the dedicated distribution of NGF in different forms using single photon emission computerized tomography(SPECT).DESIGN: It is a randomized controlled study with New Zealand rabbits as subject.SETTING: An affiliated hospital of Nanjing Medical College.MATERIALS:The trial was conducted in Nanjing Senke Medical Company and the Nuclear Medicine Department of the First Hospital of Nanjing Medical College from June 2003 to May 2004. The subjects were 19 New Zealand rabbits of either sex, weighting (2.0 ±0. 2) kg, from Anlimo Technology Company. [99Tcm] -NGF(labeling rate 98.9%, purity 99.7% )was made by Senke Medical Company. Liposome was provided by Shengyang Pharmaceutical College. Urethine solution(200 g/L) was from Pharmacy Department of NanJing Medical Collage.METHODS: [99Tcm]-NGF was encapsulated in liposome and was treated as the following: The liposomesA containing 1.48 × 108Bq[99Tcm] -NGF were injected into rabbits and its distribution percentage was analyzed with SPECT. The same amounts of[99Tcm] -NGF and[99Tcm] -NGF-ordinary liposomesB were treated in the same way.MAIN OUTCOME MEASURES: The ratio of concentration and radiation percentage of NGF in brain to those in the whole body.RESULTS: The[99Tcm]-NGF encapsulated in self-made liposomeA presented high radiation in the brain. But[99Tcm] -NGF alone was almost completely excreted through urinary system and[99Tcm] -NGF encapsuled in ordinary liposomeB was mostly phagocytized by liver reticuloendothelial system.CONCLUSION: The self-made NGF-liposomeA is brain-dedicated, which set a basis for drugs to pass through BBB.