DNA Mutational Analysis ; Drug Resistance ; genetics ; Genetic Variation ; Glucocorticoids ; pharmacology ; Humans ; Mutation ; Receptors, Glucocorticoid ; classification ; genetics ; physiology

Objective To evaluate whether the MDR1 exon 26 genotype is associated with the dosage of FK506 and acute rejection in renal transplant patients. Methods One hundred and six patients who administrated FK506 after renal transplantation were retrospectively analyzed. The patients were investigated for MDR1 genotype determined by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) assays. According to the genotype, the patients were divided into 3 subgroups, wild homozygous-CC, mutant homozygous-TT and heterozygous-CT. At 3rd, 6th and 12th month after renal transplantation, enzyme-linked immunosorbent assay (ELISA) was used to measure the FK506 whole blood concentration. Whole blood FK506 concentration per dose/kg/day and acute rejection within one month postoperation were compared among 3 subgroups. Results Of the 106 patients, 32 (30.2 %), 44 (41.5 %) and 30 (28.3 %) were carriers of CC, CT, TT in MDR1 exon 26, respectively. The whole blood FK506 concentration per dose/kg/day for patients with CC was remarkably lower that that of the patients with either CT or TT (P

Objective To assess the value of endoluminal convex array probe combined with high frequency linear array probe in diagnosis of internal carotid artery occlusion, and to improve the diagnostic accuracy rate of ultrasonography. Methods Patients suspected with ischemic cerebrovascular disease (n=490) were examined by carotid ultrasound. First, suspicious patients with carotid artery occlusion were selected using high-frequency linear array probe, then screen results were confirmed by endoluminal convex array probe and compared with previous results in those patients. Results Sixty eight patients,who were suspected of internal carotid artery occlusion,were screened out by high-frequency linear array probe. Results were re-examined by endoluminal convex array probe, 42 patients were diagnosed as internal carotid artery occlusion, 19 patients were diagnosed as severe stenosis as the internal carotid artery and 7 patients were diagnosed as ca?rotid artery tortuosity with no significant stenosis. Conclusion Endoluminal convex array probe is an effective complement for internal carotid arterial inspection when it combines with high-frequency linear probe. It can improve the diagnostic accu?racy rate of degree and the range in internal carotid artery occlusion and stenosis, therefore reduce chances of misdiagnosis.

BACKGROUND: Adhesion molecules are closely associated with inflammation. Inflammation due to white blood cell (WBC) infiltration and free radical injury following brain ischemia are believed to be important factors contributing to the pathogenesis of multi-infarct dementia.OBJECTIVE: To determine the level of serum adhesion molecules and free radicals in patients with multi-infarct dementia to explore the relationship between their levels and multi-infarct dementia.DESIGN: A case-control trial.SETTING: Department of Neurology, Second Affiliated Hospital of Jilin University.PARTICIPANTS: Totally 82 patients with multi-infarct dementia were hospitalized in the Department of Neurology, Second Affiliated Hospital of Jilin University between January 2000 and December 2004. These patients included 32 cases of mild dementia, 21 of moderate dementia, and 29 of severer dementia. The normal controls were 23 concomitant healthy volunteers who came for routine physical examination.METHODS: Enzyme-linked immunosorbent assay (ELISA) and electron spin resonance were used to determine the level of serum soluble intercellular adhesion molecules (ICAM) and vascular cell adhesion molecules (VCAM), as well as oxygen free radical concentration in the normal controls and patients with multi-infarct dementia, and the association between the severity of the illness and the levels of adhesion molecules and oxygen free radicals was analyzed.MAIN OUTCOME MEASURES: Serum levels of ICAM-1 and VCAM-1 and oxygen free radical concentration in the two groups.RESULTS: Totally 82 patients with multi-infarct dementia and 23 healthy controls were included in this study and all enter the result analysis. In multi-infarct dementia patients, the serum levels of soluble ICAM-1 and VCAM-1 and oxygen free radical concentration [(469.00±76.33), (196.00± 45.91) and (1 103.30±98.96) μg/L, respectively] were significantly higher than those of the control group [(601.00±76.30), (4.018±1.656), and (1.295±0.718) μg/g, respectively, t=5.517-6.754, P ＜ 0.01], and the 3 indices were positively correlated with the severity of dementia (r=0.659 4,r=0.697 2, r=0.649 4, respectively, P ＜ 0.05); serum ICAM-1 and VCAM-1 levels were positively correlated with the concentration of oxygen free radicals (r=0.714 7, r=0.732 4, respectively, P ＜ 0.01).CONCLUSION: Serum ICAM-1, VCAM-1 and oxygen free radicals might be implicated in the pathophysiological development of multi-infarct dementia, and their levels increase in parallel with the severity of dementia.

Objective To evaluate the therapeutic effects of autolngous bone marrow mesenchymal stem cells transplantation in the early phase of acute renal ischemia-reperfusion(I/R)injury in rabbits and their impact on tubular epithelial cell apoptosis.Method Bone marrow mesenchymal stem cells(MSCs)were isolated and cultured in vitro.Thirty rabbits were divided randomly into transplanted group,control group and sham-operated group,10 in each group.Rabbits of transplanted group were induced acute renal I/R injury by clamping bilateral renal arteries for 90 minutes,then transplanted MSCs labeled with BrdU immediately after the resumption of circulation in kidney.The rabbits of control group were induced acute renal I/R injury and then infused with saline instead.The sharn operated rabbits were transplanted autologous MSCs labeled with BrdU after sham operation.Fortyeight hours after transplantation,all rabbits were sacrificed.Renal functional and structural damage were evaluated.MSCs derived BrdU positive cells were determined by immunostalning.Tubular epithelial cell apoptosis was arialyzed by usign TUNEL.The SPSS version 11.3 Software was used for statistical analysis.ANOVA were used to ana lyze the data of renal function and tubular epithelial cell apoptesis.Data of renal structure darnage were analyzed by using the nonparametric Kruskal-Wallis test.Results Compared with control group,the renal functional and struc tural darnage were significantly ameliorated(24 h,BUN:F=7.483,Ser:F:15.091;48 h,BUN:F:17.741.Scr:F=61.865;P<0.05),tubular epithelial cell apoptods Wsa reduced significantly in transplanted group(F=135.495,P<0.01).MSCs derived BrdU positive cell were detected in the renal tissues of transplanted rabbits.Conelusiu Autologous bone marrow mesenchymal stern cells transplantation is curative for acute renal I/R injury in rabbits.Bone marrow MSCs might ameliorate renal I/R injury by reducing tubular epithelial cell apoptosis.

Objective To determine the effect of Vibrio vulnificus cytolysin (VVC) inducing ap-optosis in human umbilical vein endothelial cell(HUVEC) and its possible mechanism. Methods The en-tire vvhA gene that encoding VVC from V. vulnificus strain GTC333 was amplified by PCR and sequenced af-ter T-A cloning. E. coli BL21DE3pET-42a-vvhA, a prokaryotic expression system of the vvhA gene, was then con-structed. Ni-NTA affinity chromatography was applied to purify the target recombinant protein rVVC, and SDS-PAGE plus Bio-Rad Agarose Image Analyzor were used to measure the output of rVVC and to determine the purity of rVVC extract. The activity of rVVC dissolving rabbit erythrocytes was detected by hemolysis test. DPNH chromotometry and TphBNa chromotometry were performed to examine the contents of LDH and K+ in the supernatants of rVVC-treated HUVEC cultures, respectively. The effect of rVVC inducing apepto-sis of HUVEC was detected by flow cytometry, rVVC was labeled with FITC and the location of FITC-labe-ling rVVC in HUVEC was observed by laser canfocal microscopy. Results The cloned whA gene had 96.09% and 98.26% similarities of nucleotide and amino acid sequences compared to the corresponding se-quences in GenBank. rVVC, with a dosage of 1 μg/ml, could dissolve rabbit erythrocytes (P<0.01). 10 μg/ml rVVC was able to promote the increases of K+ content (P<0.01) but no change of LDH content could be found in the cell supernatants. HUVEC was apoptotic after the cell was treated with 1～100 μg/ml of rVVC for 2 h. In the 5～240 min duration of co-incubation of FITC-labeling rVVC and HUVEC, the rV-VC gradually moved from surface to inner side of the membrane and then entered the cytoplasms. When FITC-labeling rVVC treated HUVEC for 30 min, most of the rVVC was found to be intracellular location. Conclusion rVVC has cytolytic activity. VVC has an ability to enter HUVEC and causes injury of HUVEC via inducing apoptosis, which may be the major pathogenic mechanism of VVC.

Objective To investigate the factors affecting pulse transit time during anesthesia. Methods Sixty cases of gastrointestinal surgery under general anesthesia were investigated.The pulse transit time (PTT),BP and HR were measured at the time before and after intravenous anes-thesia induction,and at the time before and after the injection of vasoactive drugs,respectively. Results Compared to at the time before injection,HR,SBP and DBP increased significantly,and PTT decreased significantly (P <0.01 )at the time after injection of atropine and ephedrine.Com-pared to at the time before injection,HR and PTT increased significantly,while SBP and DBP de-creased significantly (P < 0.01 )at the time after injection of nicardipine.While HR and PTT de-creased significantly,and SBP,DBP increased significantly (P <0.01 )with the injection of pheny-lephrine.Obvious negative correlation between SBP and PTT was observed before and after induc-tion;meanwhile,only weak correlation between DBP and PTT was observed,and no obvious correla-tion between HR and PTT was detected.SBP and PTT were well negatively correlated with the injec-tion of atropine,ephedrine,nicardipine or phenylephrine.Conclusion All the PTT changes during anesthesia were found to be negatively correlated with the systolic blood pressure.

Objective To investigate the distribution of ompA gnne in 15 Chinese reference standard strains belonging to 15 serogroups of Leptospira interrogate, and to express recombinant OmpA ( rOmpA ) and to identify immunogenicity and immunoprotection of rOmpA. Methods Genomic DNAs from different leptospiral strains were extracted by phenol-chloroform method. Entire ompA gene fragments from the strains were amplified by PCR and then sequenced after T-A cloning. A prokaryotic expression system of ompA gene from L. interrogans strain 56601 was constructed, and the expression and yield of rOmpA were determined by SOS-PAGE plus Bio-Rad Agarose Image Analyser. Rabbits were immunized with rOmpA for obtaining antiserum, and immunodiffusion test was used to measure the antiserum's titer. Western blot assay was performed to determine the immunoreaetivity of rOmpA with the antiserum against rOmpA and antiserum against whole cell of L. interrogans strain 56601, while mi-croscopic agglutination test (MAT) was applied to detect the cross agglutination to the 15 L. interrogans strains. A leptospire adhering cell model and a leptospire infecting guinea pigs model were used to determine the adhesion-bloc-king effect of rOmpA antiserum and immunoprotection of rOmpA. Results All the 15 L. interrogans strains, but not L. biflexa strain Patoe Ⅰ , had sequence conserved ompA genes. The yield of rOmpA was approximate 20% of the total bacterial proteins, rOmpA could induce rabbits to produce antibody and immunodiffusion titer of the anti-serum was 1:4. Both antisera against rOmpA and against whole cell of L. interrogans strain 56601 were able to pro-duce positive Western blot signs to rOmpA, and the former offered 1 : 20-1 : 320 MAT titers to the 15 L. interrogans strains. 1: 10-1:160 dilutions of rOmpA antiserum could efficiently block L. interrogans strain 56601 adhering to J774A. 1 cells, and 100 μg and 200 μg rOmpA displayed 50.0% and 75.0% immunoprotective rates in the infee-ted guinea pigs. Conclusion ompA gene only exists in genomes of different pathogenic L. interrogans serogroups. rOmpA has relatively stronger antigenicity, cross immunoreactivity and certain immunoprotection, implying that this recombinant protein may be used as a candidate antigen for developing universal genetic engineering vaccine of L. interrogans.

Objective To observe the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) on patients with type A thoracolumbar fractures treated by pedicle screw fixation via dorsal-approach and vertebral body bone graft.Methods A retrospective case-control study was conducted to analyze the clinical data of 64 patients with type A thoracolumbar fractures treated from June 2012 to July 2015.The patients were divided into control group (32 cases) and research group (32 cases) according to the random number table.There were 22 males and 10 females aged (48.2 ± 11.2)years old in control group.The injury was located at T11 in 2 cases,T12 in 4,L1 in 9,L2 in 7,L3 in 7 and L4 in 3 in control group.There were 24 males and 8 females aged (50.7 ± 11.4) years old in research group.The injury was located at T11in 2 cases,T12in 4,L1in 8,L2 in 8,L3 in 7 and L4 in 3.The control group was treated with posterior pedicle screw reduction,internal fixation and allograft cancellous bone grafting through pedicle of vertebral arch.The research group was treated with rhBMP-2 (mixed with cancellous bone implants) on the basis of the control group.The time of operation,the amount of bleeding during operation,the volume of postoperative bleeding,the amount of postoperative drainage,the recovery of the injured vertebra,the Lane-Sandhu score,the time of fracture healing,and the postoperative complications were compared.Results There was no difference between two groups on operation time,amount of bleeding during operation or amount of postoperative drainage (P ＞ 0.05).No difference existed in two groups about Cobb angle and injured vertebral leading edge height at one week after operation (P ＞ 0.05).At the time of last follow-up (12 months),in research group,Cobb angle was lower while injured vertebral leading edge height was much higher than that of control group (P ＜ 0.05),both of which were better than before (P ＜ 0.05).At 12 months after surgery,the lost Cobb angle of [(2.0 ± 0.7) °] and lost height of injured vertebral leading edge [(3.2 ± 1.0) ％] were smaller than those in control group [(5.6 ± 1.7) ° and (6.8 ± 2.4) ％ respectively] (P ＜ 0.05).The Lane-Sandhu score and healing time of fracture in research group were (8.9 ± 0.8) points and (6.9 ± 0.9) months,which were better than that (6.8 ±0.8)points and (8.4 ± 1.6)months in control group respectively (P＜0.05).The complication rate of research group was lower than that of control group,with no significant difference (P ＞ 0.05).Conclusion Compared with simple vertebral bone graft,posterior thoracolumbar vertebrae pedicle screw fixation with vertebral bone graft and hBMP-2 treatment for type A thoracolumbar fractures can restore vertebral stability,shorten the time of fracture healing and reduce the incidence of complications.

40 years old in 6 Uygur communities of Urumqi. The multi-phasic stratified cluster sampling method was adopted. Finally 191 men with high risk were selected as samples in the study and classified into two groups according to oral glucose tolerance test(OGTT)results, anthropometric and biochemical indicators. Results The prevalence of type 2 diabetes and IGR was 17.39% and 7.56% respecfively in the Uygur nationality. Waist-hip ratio, abdominal and waist circumference,the level of HbA1c and serum insulin of male diabetic patients were significantly higher than those of male IGR potieuts. But another four indicators of blood lipid and three indicators of renal function were not significantly different between two groups. Conclusion The indicators releted to abdominal obesity may be sensitive to diabetes and valuable to screen diabetic patients for giving effective health education and behavioral intervention in high risk population.