We compared the total errors observed from both precision and accuracy validation experiments with defined medically allowable errors for each quantitative test,and used Westgard's method evaluation decision chart for initial judgement of the acceptability of method validation performance. Moreover,we need to judge the acceptability of other method validation parameters,so that we could integratedly assess and judge whether the testing system is acceptable to use in the laboratory.

Animal Feed ; Animals ; Gynostemma ; Male ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Swimming

Objective: To investigate the concentrations of secreted protein acidic and rich in cysteine(SPARC) in the serum and urine of patients with IgA nephropathy and its expression in the kidney tissues. Methods: The concentrations of SPARC, tumor necrosis factor-a (TNF-a),interleukin 1β (IL-1β), and interleukin 6 (IL-6) in the serum and urine were measured with enzyme-linked immunosorbent assay(ELISA). The contents of SPARC protein in the culture medium of human mesangial cell (HMC) and human renal tubular epithelial cell (HKC), which had been treated with IL-6, were determined by ELISA. The expression and distribution of SPARC in IgA nephropathy and normal kidney tissues were observed by immunohistochemistry assay. Results: The concentrations of SPARC in serum and urine of IgA nephropathy patients were higher than those of the normal control subjects ([2.43±±1.22] μg/ml vs [0.69±0.21] 2μg/ml, [7.73±2.81] μg/ml vs [1.17±1.03]ρg/ml, P<0.01). The serum levels of TNF-α, IL-1β and IL-6 in IgA nephropathy group were significantly higher than those in the control group (P< 0.05); the urinary levels of TNF-α and IL-6 in IgA nephropathy group were also higher than those in the controls (P<0.01). SPARC protein secreted by HKC was higher than that by HMC (P<0.01). SPARC was weakly positive in normal distal cortical tubules. SPARC protein expression in tubular epithelial cells of IgA nephropathy patients was obviously higher than that of the normal controls. Conclusion: The secretion of SPARC by renal tubular epithelial cells is increased in patients with IgA nephropathy, which results in elevation of serum SPARC and may have a protective feedback inhibitory effect on HMC proliferation.

Objective: To observe the changes of Bcl-xl expression in hippocampus of the rats with the forced-swimming and the changes of Bcl-xl after poststress. Methods: The protocol was established with the forced-swimming as the chronic stress model. Open-field test was executed to measure the behaviors of the rats. Immunohistochemistry was used to observe the changes of Bcl-xl expression in the hippocampus. Results: Compared to the control group,the expression of Bcl-xl in CA3 region of the hippocampus and dendate gyrus(DG)of the rats was decreased morphologically. With the computerized image analysis, the gray degree increased significantly (P

[Objective]To observe the biological properties of a novel collagen/hydroxyapatite composite scaffold in vitro and to evaluate the possibility of application being used in tissue engineering for osteochondral repair.[Method]The scaffolds were constructed of collagen I and hydroxyapatite.The pore size and interpores of the scaffold were observed by scanning electronmicroscopy(SEM).The porosity was measured by liquid displacement method.Rabbit bone marrow stromal cells(BMSCs) were isolated and amplified,then inoculated onto the scaffold.By SEM scanning,the condition of the cells adhering onto the scaffold was observed.The proliferation of the cells on the scaffolds was examined using MTT method,and the growth curve was drawn.[Result]The scaffold possessed high porosity and proper pore size.The pore diameter of the collagen layer was about 90?m,the pore diameter of the HA layer was about 120?m,and the overall porosity of the composite scaffold was 75%.The proliferation of the cells on the scaffold was good.[Conclusion]The novel collagen/hydroxyapatite composite scaffold possesses desirable pore structure and good biocompatibility,and it can be used in tissue engineering for osteochondral repair.

Objective　To investigate the pattern of human neutrophil degranulation mediated by Fc alpha receptors. Methods　IgA immune complexes (IgA IC) and specific monoclonal antibody were used to stimulate neutrophil degranulation. Lactoferrin released by stimulated neutrophils was quantitatively determined. Results　Monoclonal antibody and IgA IC could both induce neutrophil lactoferrin release. However, monoclonal antibody induced rapid and strong degranulation whilst the effect of IgA IC was much weaker. Conclusion　The pattern of neutrophil degranulation induced by IgA IC and monoclonal antibody is different.

Objective To study the relationship between GAT-1 and neuron injury in focal cerebral ischemia/infusion model.Methods Focal cerebral ischemia/reperfusion models were established in rats by thread embolic, then the brain tissues were taken at 3 h, 6 h,12 h, 24 h and 3 d after cerebral ischemia/reperfusion.The number of GAT-1 positive neurons were calculated and the average optical density were detected by immunohistochemical technique in different time points after cerebral ischemia/reperfusion, compared with the normal group and the sham operated group.Results The expression of GAT-1 was up-regulation obviously at 3 h after cerebral ischemia/reperfusion( P

Objective To investigate the changes of expression of Fas-associated proteins named Fas-associated death domain protein(FADD)and death-associated protein(Daxx)in the ischemic penumbra following transient focal cerebra ischemia in rats.Methods ①Adult male Sprague-Dawley rats were randomly divided into the sham-operated group and the cerebral ischemia model group.Rats underwent right middle cerebral artery occlusion(MCAO)for 2 h and reperfusion for 1,3,6,12 and 24 h using an intraluminal suture technique.The expression of FADD and Daxx mRNA and protein were measured with methods of immunohistochemistry.Western blot and reverse transcription-polymerase chain reaction(RT- PCR)respectively were used in the ischemic penumbra of rats.②Double-label fluorescence confocal laser scanning microscopy(CLSM)was performed to monitor FADD and Daxx intracellular location before and after ischemia.Results RT-PCR,Immunohistochemistry,Western blot experiments indicated that a very low level of FADD mRNA and protein were detected in the cerebral cortex of sham rats.The expression level both of FADD mRNA and protein increased significantly at 3 h after reperfusion,peaked at 12 h,then declined markedly at 24 h in the ischemic penumbra of model rats.RT-PCR,Immunohistochemistry indicated that a relatively high level of Daxx mRNA was detected in the cerebral cotex of sham rats.The expression level of Daxx mRNA increased significantly at 3 h after reperfusion and persisted to 24 h at a high level,whose protein had a same change of expression level in the ischemic penumbra of model rats. Immunofluorescence double-staining laser scanning by CLSM showed that the immunoreactivity of FADD was located in cytoplasm,and the intracellular translocation of the immunoreactivity of Daxx from nucleus to cytoplasm was monitored by measuring the green fluorescence after ischemia.Conclusion The transient upregulation of FADD and the persistant high level of expression of Daxx may contribute to neuronal apoptosis following cerebral ischemia/reperfusion.

60 years old). 43 cases of antiepileptic drugs combination accounted for 10% and the therapeutic plasma concentration of 27 cases deviated from normal range(62.8%). CONCLUSION:Results of plasma concentration monitoring provide an important basis for clinical drug use. Monitoring data and other clinical index can promote rational use of antiepileptic drugs.

Objective To establish more simple and effective rat limbs transplantation. Methods Taking Wister rat as donatur,SD rat as recipient,and transplanting Wister rat rear limbs (right or left) to SD rat.The experiments were carried out in two group,the traditional group (after mutilation of SD rat limbs, transplanting the similar side Wister rat rear limbs to SD rat by fixing bone,anastomosing nervi vasorum, suturing muscle and skin) and the improvement group (retaining SD rat limbs,and fixing Wister right limbs on the left inner side of SD rat limbs or fixing Wister left limbs on the right inner side of SD rat limbs by suturing muscle and skin. Then only anastomoing femoral artery and femoral vein, and not anastoming nervi, not fixing bone). Recording operation time and weight change after operation. Microsurgery usual care Recording host drink and food and weight. Observing implant change. Results Seventy-four operations were accomplished successfully. Graft rejection reappear stably. All 74 rats survived over 14 days after transplantation. The operation time of traditional group and imrovement group were (125±40) min and (70 ±21) min respectively. Decreased body weight of traditional group and improvement group were (3.78± 1.09)and(2.05±0.90) g respectively. After 3 days, the weight of improvement group rat begin to increase, however, traditional group rat always dcrease in one week. Operation achievement ratio of traditional group and improvement group were 38%, 90% respectively. Improvement group is better than traditional group in above 4 section. Conclusion The method is more simple,applicable and requires shorter time. Less trauma to rats and rats recover quickly. It can be used in establishing rat limbs transplantation acute rejection model.