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Objective To investigate platelet aggregation on glass surface under physiological flow condition. Methods The polydimethylsiloxane (PDMS)-glass microchannel chips were fabricated by soft lithography. Anti-coagulant human peripheral whole blood was flowed through the microchannel chip at flow shear rate of 300 s-1 and 1 500 s-1, respectively. The fluorescence images of platelet aggregates formed on glass surface at the bottom of the microchannel were captured after 150 s using an inverted fluorescence microscope. The number of platelet aggregates, average size, surface coverage and average fluorescence intensity were quantified by image analysis. The glass surface was treated with oxygen plasma, BSA blocking or collagen modification to establish different surfaces for platelet aggregation. The hematocrit (Hct) of blood sample was adjusted, and the whole blood was treated with different anti-platelet agents. The platelet aggregation on glass surface was observed under the above experimental conditions. The platelet aggregations in healthy people and diabetic patients were also analyzed. Results Under the flow condition, platelet aggregation on glass surface was three-dimensional. Platelet aggregation was dependent on wall shear rate, the hydrophilicity of glass surface and Hct, and was mainly regulated by GPIIb/IIIa-fibrinogen and ADP-P2Y12 receptor pathways. The aggregation of platelets on the glass surface could also reflect the high activity of platelets in diabetic patients. Conclusions At the flow conditions of 300 s-1 and 1 500 s-1, platelet aggregation on glass surface is related to flow rate, protein adsorption, platelet related receptors and platelet activation state. In this study, a new model for microfluidic platelet function analysis without additional adhesion protein modification was established, and it could be used for clinical evaluation of platelet function.
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Objective:To understand the laboratory characteristics for the diagnosis of immunoglobulin G4-related disease (IgG4-RD).Methods:The clinical data of 28 patients with IgG4-RD and renal damage (IgG4-RKD) diagnosed in our hospital from January 2017 to May 2019 were retrospectively analyzed. The correlation between serum IgG4 concentration and clinical features as well laboratory test results was analyzed. The 28 patients were divided into two groups: high serum IgG4 concentration group and normal serum IgG4 concentration group. The serum creatinine value, erythrocyte sedimentation rate, IgG concentration, IgA concentration, complement C3, C4 concentration, peripheral blood eosinophils, hemoglobin, IgG4/IgG and other related parameters were compared between the two groups. SPSS 20.0 statistical software was used for analysis. The two groups of measurement parameters were compared between groups by independent sample t test, non-normal measurement parameters were compared between groups by Mann-Whitney U test analysis, and the correlation between patients' IgG4 and each detection parameter was analyzed by Spearman correlation analysis. Results:Among the 28 patients, 17 were male and 11 were female, with an average age of (62±14) years. The serum IgG4 concentration increased in 75% of the patients ( n=21), with an average value of 3.01(1.41, 7.52) g/L, the serum IgG concentration increased in 64.3% of patients ( n=18), with an average value of 18.91 (12.88, 24.88) g/L, and the complement C3 decreased in 50% of the patients ( n=14), with an average value of(0.77±0.28) g/L. IgG4 was positively correlated with IgG ( r=0.422, P=0.025), IgG4/IgG ( r=0.951, P<0.01), ESR ( r=0.543, P<0.01) and peripheral blood eosinophils ( r=0.487, P<0.01), but negatively correlated with complement C3 ( r=-0.431, P=0.022) and C4 ( r=-0.504, P<0.01) levels. There were significant differences in IgG ( Z=-2.255, P=0.023), IgG4/IgG ( Z=-3.793, P<0.01), C3 ( t=7.380, P<0.01) and ESR ( t=-2.195, P=0.037) between the elevated IgG4 group and the normal group. Conclusion:Serological characteristics of IgG4-RKD combined with clinical manifestations may be able to diagnose IgG4-RKD in early stage.
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Objective To assess the efficacy of different sequential therapy regimens according to the theory of cell cycle on adriamycin-induced nephropathy (AIN) rats. Methods SD rats were randomly divided into five groups:control group (n=8),AIN model group (n=8),MP+ CsA+MMF group (n=8),MP+CsA+CTX treated group (n=8) and MP+FK506+Rapa group (n=8).The levels of 24-hour urinary protein,serum total protein (TP),albumin (Alb),cholesterin (Chol),triglyeride (TG),serum urea nitrogen (BUN),serum creatinine (Scr) were measured.The renal pathological changes were observed by light microscope.The expressions of nephrin and podocin were analyzed by immunohistochemistry and real-time PCR.The expression of CTGF was detected by Western blotting. Results (1)Compared with control group,the levels of 24-hour urinary protein in AIN model group were obviously increased at 2nd,4th,8th and 12th week (all P＜0.01).The level of 24-hour urinary protein were obviously decreased in treatment groups at 8th and 12th week than those in AIN model group (all P＜0.05). (2)The levels of TP and Alb were significantly lower in AIN model group than those in control group (all P＜0.01),and the levels of TG and Chol were significantly higher in AIN model group than that in control group (all P＜0.01).The levels of TP and Alb were significantly higher and the levels of TG and Chol were significantly lower in treatment groups than those in AIN model group (all P ＜0.05). (3)The results of immunohistochemistry indicated the expressions of nephrin and podocin in treatment group rats were obviously higher than those in AIN model group (all P＜0.01). (4)The results of Western blotting indicated the expression of CTGF in treatment group rats was higher than that in AIN model group (P＜0.05).The effect of inhibitting fibrous degeneration in MP +FK506 +Rapa group was more greater than other treatment groups. Conclusions Sequential combined regimens according to the cell cycle can improve the pathological change in adriamycin-induced nephropathy rats,reduce the urine protein,increase the levels of TP and Alb,decrease the levels of TG and Chol,increase the expression of nephrin and podocin,and ameliorate kidney fibrosis.