Objective: To evaluate the effect of pretreating donor liver with salvianolate on biliary injury after orthotopic liver transplantation (OLT) in rats, and to discuss the underlying mechanisms. Methods: According to the preservation solutions of the liver grafts, the recipients rats were allocated in three groups: UW solution group, salvianolate group (UW solution + 0.4 mg/ml salvianolate), and saline group (UW solution + saline). In experiment 1, the recipient rats underwent arterialized OLT with biliary extra-drainage, and the bile flow and bile salt secretion were recorded at 1, 6, and 24 h post-transplantation. In experiment 2, the recipient rats underwent arterialized OLT, and the serum alkaline phosphatase, bile salt, total bilirubin, and the incidence of biliary complications were examined at 2 weeks post- transplantation. Meanwhile, the mRNA and protein expression of bile salt export pump (BSEP) in the liver samples were also detected using real-time RT-PCR and Western blotting analysis, respectively. Results: Compared with the UW solution and saline groups, salvianolate group exhibited significantly increased bile flow and bile salt secretion at 1, 6, and 24 h post-transplantation (P<0. 05). At 2 weeks post-transplantation, salvianolate group had significantly lower serum alkaline phosphatase, bile salt, total bilirubin(P<0. 05), and incidence of biliary complications(P<0. 01), and significantly higher expression of BSEP mRNA and protein compared with the other two groups (P<0. 05). Conclusion: Salvianolate addition in UW preservation solution can ameliorate biliary injury and reduce the incidence of biliary complications after OLT in rats, which might be associated with increased expression of BSEP in liver graft.

Objective: To observe the effect of salvianolate on the maturation and immunologic functions of mouse bone marrow-derived dendritic cells (BMDCs). Methods: Mouse BMDCs were cultured with GM-CSF and 1L-4 for 5 days to obtain immature DCs (imDCs),then the imDCs were cultured in the presence of LPS (1 μg/ml) or LPS (1 μg/ml) plus salvianolate (50, 100 and 200 μg/ml) for 48 h to obtain DCLPS , SVL , SVM and SV H. The immunophenotypes and antigen phagocytosis capabilities of imDCs, DCLPS, SVL, SVM and SVH were analyzed by flow cytometry and FITC-Dextran intake. The IL-12 p40 contents in the supernatants were determined by enzyme linked immunosorbent assay (ELISA). The stimulatory activities of imDCs, DCLPS, SVL ,SV M and SVH on allogeneic T cells in mixed lymphocyte reactions (MLR) were tested by incorporation of 3H-TdR; the contents of IL-10,IL-2 and IFN-γ in the supernatants of MLR were examined by ELISA. Results: Compared with DCLPS,SVL,SVM and SVH had lower expression of MHC-II, CD40,CD80 and CD86 molecules, secreted less IL-12 p40 in the supernatant, and exhibited stronger antigen phagocytosis capability; moreover, SVL ,SVM and SV H showed significantly weaker activity in stimulating proliferation of allogeneic T cells(P<0.05,0.01). Compared with DCLPS, SV L, SVM and SVH groups had significantly decreased IL-2 and INF-γ levels in MLR supernatants(P<0.05), but there was no significant difference in the secretion of IL-10. Conclusion: Salvianolate (50-200 μg/ml) can inhibit the maturation of mouse BMDCs in vitro, and thus has a negative effect on immunologic function of BMCDs.

In order to establish an UPLC-MS method for determination of twelve active compounds in Qili Qiangxin capsules including astragaloside, calycosin-7-0-glucoside, ginsenoside Rb1, ginsenoside Re, ginsenoside Rd, ginsenoside Rg1, ginsenoside Rf, periplocin, periplocoside H1, hesperidin, narirutin, isoquercitrin, the chromatographic separations were performedon a Phenomenex UPLC Kinetex C18 column (2.1 mm x 100 mm, 2.6 microm) with gradient elution of acetonitrile and 0.1% aqueous formic acidat a flow rate of 0.4 mL x min(-1). The temperature was set as 40 degrees C and injection volume was 5 microL. The monitoring of all analytes was achieved under the negative ionization mode with TOF-MS and TOF-MS/MS method. The twelve analytes showed good linearity (R2 > 0.9990) within the test ranges, the average recoveries were 98.0%-102%, respectively, and the RSD were less than 3.9%, respectively. The established method is simple, rapid, and sensitive, and can be used for quality control of Qili Qiangxin capsules.
Capsules ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods

OBJECTIVETo explore clinical significance of rotational axis of distal femur on Chinese adults in total knee arthroplasty (TKA).

METHODSThere were 86 Chinese adults (106 normal knees) including 47 males (53 knees) and 39 females (53 knees), 54 knees were on left and 52 on right. The CT scan was employed in the distal femur. The scan direction was aligned to be on the plane perpendicular to the mechanical axis of the femoral. The CT images of cross sections across lateral and medial femoral epicondyle were moved to personal computer,lateral angle between anterior posterior line (APL) and surgical transepicondylar axis (STEA) (ATA),lateral angle between posterior condylar line (PCL) and APL (APA), angle between perpendicularity of APL and PCL (A-PA), posterior condylar angle (PCA), condylar twist angle (CTA), angle between clinical transepicondylar axis (CTEA) and STEA (CSA) were measured. These values were divided into different groups according to gender and side, the values of CTA, PCA, A-PA, angle PT (varus of tibia plateau), constant 3, ATA, APA and constant 90° were compared by statistically. A-PA and PCA, and CTA were analysed statistically with the liner regression, the relationship among CTEA, STEA ,PCL, APL and PLP were performed to assess by liner regression.

RESULTSATA was (89.79 ± 1.22)°, APA was (84.84 ± 1.83)°, A-PA was (5.16 ± 1.83)°, PCA was (4.80 ± 1.23)°, CTA was (8.23 ± 1.40), CSA was (3.45 ± 0.68)°. All the parameters had no differences on sex and side,but CSA had difference on male and female. There was no difference among angle PT, PCA, A-PA. There was significant difference in CAT, constant 30 and angle PT, PCA,A-PA. There was no difference between ATA and constant 90°, but there was difference between APA and constant 90°. There was relativity between PCA and CTA, and also PCA and A-PA, CTA and A-PA. There was significant relativity between STEA and CTEA, between STEA and APL, between STEA and PCL, and also between APLP, APL and PCL, but there was no significant relativity between PCL and CTEA.

CONCLUSIONTKA for Chinese, the section of femoral posterior condyle should be external 5° to obtain the optimum rotational orientation. The property is different entirely between STEA and CTEA, the rotational alignment is not performed according to parallel to the CTEA in distal femur. Among STEA, APL, PCL, the STEA is the most reliable mark ofrotational alignment of the distal femur, and the PCL is the less reliable mark.

Adult ; Arthroplasty, Replacement, Knee ; Female ; Femur ; anatomy & histology ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Rotation ; Tomography, X-Ray Computed

Objective To investigate the effect of recombinant human erythropoietin(rh-EPO) on the nerve regeneration of adult rats sciatic nerves. Methods Tirty-six healthy male Wistar rats were involved and left sciatic nerve repaired model was used.The experimental rats were divided randomly into two groups:the EPO group and the control group,18 rats in each group.rh-EPO 3 000 U/kg was injected daily into the abdominal in EPO group,and normal saline was injected into the abdominal every day after operation in control group.On 4 and 8 weeks after operation,these items were determined,the sciatic function index (SFI),biomechanics examination,histological observation,electrophysiological examination,myelinated fibers density and sectional area measurement.Results On weeks 4 after operation,the SFI of EPO group and control group were-65.26 ± 3.42 and-70.83 ± 4.12,respectively,the maximum tensile resistance were (3.86 ± 0.29)N/mm2 and (3.38 ± 0.21 )N/mm2,the delayed ratio of latency of motor nerve were 2.34 ± 0.23 and 2.78 ± 0.29,and the recovery ratio of wave amplitude were 0.23 ± 0.05 and 0.14 ± 0.03 respectively.On eight weeks after operation,the SFI of EPO group and control group were-51.34 ± 2.98 and-57.23 ± 4.86,respcetively,the maximum tensile resistance were (4.67 ± 0.36) N/mm2 and (4.13 ± 0.32) N/mm2,the delayed ratio of latency of motor nerve were 1.32 ± 0.15 and 1.62 ± 0.21,the recovery ratio of wave amplitude were 0.41 ± 0.09 and 0.26 ± 0.07,the nerve fibers cross ratio were 0.57 ± 0.05 and 0.38 ± 0.03,and the recovery ratio of sectional area of myelinated fibers were 0.81 ± 0.06 and 0.58 ± 0.03,respectively.Those items in EPO group were significantly superior to those in the control group (P ＜ 0.05 ＝.Conclusion rh-EPO can promote the injured nerve regeneration and improve the recovery of their function.

Objective To observe the expression of cardiotrophin-1(CT-1) in ischemia-reinfusion cardiac muscle of rats and the effect of neuregulin-1(NRG-1).Methods The model of ischemia-reinfusion cardiac muscle of rats were prepared,35 rats were randomly divided into 4 groups:model group(n=8),NRG-1 pretreatment group(n=9),pseudo-surgery group(n=8) and normal control group(n=10).The CT-1 mRNA in the observed cardiac muscle of all groups was measured by RT-PCR and the relative amount of CT-1 mRNA were calculated,and for statistical treatment.Results The CT-1 mRNA of model group was(63.96?9.34),and it was higher than that of pseudo-surgery group(36.16?5.43)and normal control group(36.84?4.64).The significant differences were found in 3 groups(F=47.37 P

Objective To investigate the protective effect of salvianolate against bile duct injury after donation after cardiac death (DCD) liver transplantation and its clinical application prospect. Methods Sixty recipients of DCD liver transplantation were randomly divided into two groups with 30 cases in each group. Salvianolate (250mg/d) was given daily for 14 days after operation in treatment group, and the same amount of normal saline was given in control group. The therapeutic regimen of anti-rejection and anti-infection is the same between the two groups. The incidence of early graft dysfunction (EAD), and the serous levels of total bilirubin (TB), alkaline phosphatase (ALP), γ-glutamyltransferase (GGT), total bile acid (TBA) 1 month, 6 months and 12 months after liver transplantation were compared between the two groups. Also, the platelets (PLT), prothrombin time (PT), activated partial prothrombin time (APTT) and fibrinogen (FIB) were compared between the two groups at 2 weeks after operation. Results There was no significant difference in baseline parameters between the two groups (P>0.05). Compared with the control group, the incidence of EAD was decreased in treatment group, but there was no significant difference [10.0%(3/30) vs. 23.3%(7/30), P=0.166]. The serous levels of TB, ALP, GGT and TBA in treatment group were lower than those in control group 1 month, 6 months and 12 months after operation: [TB: 1 month, (28.5±17.0)μmol/L vs. (39.8±20.1)μmol/L, P=0.025; 6 months, (24.5±10.6)μmol/L vs. (33.3±16.4) μmol/L, P=0.018; 12 months, (19.8±9.5)μmol/L vs. (26.4±14.1)μmol/L, P=0.037, ALP: 1 month, (147.3±76.9)U/L vs. (187.6±70.9)U/L, P=0.039; 6 months, (163.0±61.4)U/L vs. (198.1±51.6)U/L, P=0.020; 12 months, (167.9±59.9)U/L vs. (200.2±56.2)U/L, P=0.036, GGT: 1 month, (83.9±49.5)U/L vs. (113.6±61.1)U/L, P=0.043; 6 months, (130.9±48.7)U/L vs. (169.7±77.0)U/L, P=0.023; 12 months, (154.7±45.1)U/L vs. (182.5±59.8)U/L, P=0.047, TBA: 1 month, (6.6±2.1)μmol/L vs. (8.0±2.4)μmol/L, P=0.016; 6 months, (9.5±2.2)μmol/L vs. (12.1±3.4)μmol/L, P=0.001; 12 months, (12.5±2.7)μmol/L vs. (5.6±3.8)μmol/L, P=0.001]. However, there was no significant difference in PLT, PT, APTT and FIB between two groups [PLT: (148.6±88.6)×109/L vs. (152.8±74.4)×109/L, P=0.843; PT: (12.9±1.1)s vs. (13.0±1.1)s, P=0.617; APTT: (34.6±3.7)s vs. (34.9±3.4)s, P=0.716; FIB: (3.4±0.6)g/L vs. (3.2±0.6)g/L, P=0.270, repectively]. Conclusions Salvianolate has a protective effect against bile duct injury after DCD liver transplantation, and does not increase the risk of postoperative bleeding.

Objective To establish and improve the abdominal heterotopic heart transplantation model in mice, so as to provide references for the research on organ transplantation immunology. Methods C57BL/6 mice were selected as donors and recipients for syngeneic cardiac transplantation and recipients for allogeneic cardiac transplantation; BALB/c mice were selected as donors for allogeneic cardiac transplantation. The ascending aorta and pulmonary artery of the heart of the donors were end-to-side anastomosed with the abdominal aorta and inferior vena cava of the recipients, respectively, to establish abdominal heterotopic heart transplantation models in mice. Furthermore, the techniques of preparing recipient heart before implantation, extracting and preparing donor heart, and vascular anastomosis were improved, and the surgical results and advantages were observed and analyzed. Results Preliminary experiments were performed on 50 pairs of mice. For formal experiments, syngeneic cardiac transplantation was performed on 20 pairs and the success rate was 85.0% (17/20); allogeneic cardiac transplantation was performed on 15 pairs and the success rate was 86.7% (13/15). The total success rate of abdominal heterotopic heart transplantation was 85.7% (30/35). The average time for preparing recipient before vascular anastomosis was (11.2±2.5) min, the average time for extracting and preparing donor heart was (13.6±3.3) min, and the average time for vascular anastomosis was (21.7±3.5) min. Conclusion The improved abdominal heterotopic heart transplantation model in mouse has the advantages of easy-to-perform and high success rate, which lays a foundation for the further study of transplantation immunology.

BACKGROUNDThere were some papers published in the Jonrnal of Science, December 2000 suggesting that one or more important susceptibility genes for late onset Alzheimer's disease (LOAD) were located on the long arm of chromosome 10. Linkage analysis showed maximum lod score close to D10S1225 loci, which indicated the loci might contribute to the etiology of Alzheimer's disease (AD).

METHODSFifty-nine LOAD patients and 107 controls were recruited. Apolipoprotein E (ApoE) genotypes were determined by reverse dot blotting hybridization assay. The D10S1225 was genotyped by 12% nondenaturing polyacrylamide gels electrophoresis and analyzed by silver staining. Statistical analysis was used to compare genotype and allele distributions between LOAD group and control group for ApoE and D10S1225 polymorphisms.

RESULTSApoE epsilon 4 was significantly higher in LOAD group in comparison with the control group (chi(2) = 6.530, P = 0.011). Seven different alleles of D10S1225 have been identified. The length of these gene fragments were 178 bp, 181 bp, 184 bp, 187 bp, 190 bp, 193 bp, and 196 bp, respectively. A total of 21 different genotypes were observed. There was no relationship between D10S1225 polymorphism and LOAD (chi(2) = 4.488, P > 0.05). Conclusion This study suggests that ApoE epsilon 4 is a risk factor for LOAD, however, the results indicated that there is not any possible linkage for disequilibria with a nearby AD risk gene near D10S1225.

Aged ; Aged, 80 and over ; Alleles ; Alzheimer Disease ; genetics ; Apolipoproteins E ; genetics ; Female ; Genotype ; Humans ; Linkage Disequilibrium ; Male ; Middle Aged ; Polymorphism, Genetic

Proteinase-activated receptor-2 (PAR-2) expression has been observed on numerous cell types. However, little is known about the functional expression of PAR-2 in human mast cells. In the current study, the actions of a PAR-2 agonist trans-cinnamoyl-Leu-Ile-Gly-Arg-Leu-Orn-amide (tc-LIGRLO) on tryptase release from dispersed human colonic mast cells were examined. The results showed that tc-LIGRLO was able to induce a fold increase in tryptase release over the basal level following a 15 min incubation of colonic mast cells, whereas tc-OLRGIL did not have any effect on tryptase release. The potency of tc-LIGRLO appeared greater than that of anti-IgE and calcium ionophore A23187 (CI) in induction of tryptase release. Extending the incubation time to 30 min had no significant effect on the actions of tc-LIGRLO or anti-IgE. In the time course study, it was observed that the tryptase release from mast cells induced by tc-LIGRLO started at 1 min and peaked at 3 min following incubation. The above-mentioned results indicate that tc-LIGRLO is a potent stimulus of tryptase release from human mast cells, which strongly suggests that PAR-2s are expressed in human mast cells.
Cells, Cultured ; Humans ; Mast Cells ; drug effects ; metabolism ; Receptor, PAR-2 ; agonists ; Tryptases ; metabolism