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In vitro lipolysis model has become a new and promising technique to screen and evaluate oral lipid formula-tions .This model mimics the gastrointestinal tract environment and well reflects the fate of lipid formulations in GI tract after oral administration .This review summarizes the characteristics of lipid formulations ,process of gastrointestinal digestion ,ap-plications of in vitro lipolysis model and methods for the characterization of the lipolysis process ,which provides the basis for the research of oral absorption mechanism and in vivo-in vitro correlations of lipid formulations with lipolysis model .
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Objective To understand the status of infection with multidrug-resistant organisms (MDROs) in intensive care units(ICUs),and evaluate the intervention efficacy of targeted monitoring.Methods Prospective study was adopted,patients who were admitted to ICUs in 2014-2015 were selected (January-December 2014 was as preintervention stage,January-December 2015 was as intervention stage),trend of MDRO infection before and after intervention were compared and analyzed.Results Before and after intervention,297 and 217 strains of MDROs were isolated respectively,except carbapenem-resistant Pseudomonasaeruginosa (CRPA),the isolated strains of carbapenem-resistant Acinetobacterbaunannii (CRAB),carbapenem-resistant Enterobacteriaceae (CRE),methicillin-resistant Staphylococcus aureus(MRSA),and vancomycin-resistant Enterococcus (VRE) declined after intervention.MDRO infection rate declined from 7.17 ％ before intervention to 3.88％ after intervention,infection rate of CRAB and CRE after intervention were both lower than before intervention (both P＜0.05);MDRO infection rates in general ICU and internal medicine ICU increased from 8.75％ and 7.84‰ before intervention to 4.39‰ and 2.28％ after intervention,respectively (both P＜0.05).After taking comprehensive intervention measures,compliance to prevention and control measures,such as ordering rate of doctor's advice on contact isolation for 24 hours,hand hygiene,health care workers' awareness all enhanced significantly(all P＜0.05).Conclusion Targeted monitoring and intervention measures can reduce isolation rate of MDROs in ICUs.
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Objective@#To investigate the effect of mesenchymal stem cells (MSCs) on apoptosis of breast cancer cell line MCF-7 induced by cisplatin (DDP), MSCs derived from breast cancer (BC-MSCs) or adjacent non-cancerous tissues (BN-MSCs) were isolated, cultured and identified.@*Methods@#BC-MSCs and BN-MSCs were isolated and cultured by tissue adherent method. The differentiation potential of BC-MSCs was detected by osteogenic and adipogenic induction, and cell surface markers of BC-MSCs and BN-MSCs were evaluated by flow cytometry. MCF-7 cells were co-treated with DDP and conditioned medium (CM) collected from BC-MSCs and BN-MSCs after being cultured for 48 hours, respectively. Inhibition rate of cell proliferation was evaluated by MTT. Cell apoptosis and viability were detected by MUSE cell analyzer. Cytokines in MSC-CM were detected by Luminex liquid chip. Interleukin 6 (IL-6) mRNA expressions in MCF-7 cells with different treatment were detected by RT-PCR.@*Results@#The morphology of BC-MSCs and BN-MSCs successfully isolated and cultured was uniform fibroblast-like clusters under the microscope. These cells expressed high levels of CD29 and CD44, but neither CD14 nor CD34 were detected. MSCs could also differentiate into osteoblasts and adipocytes after specific induction. After treatment with 2.5, 5, 10, 20, 40 and 80 μmol/L DDP, the inhibitory rates of proliferation of MCF-7 cells in DDP group were (17.33±2.00)%, (22.37±0.73)%, (30.77±1.23)%, (44.93±1.27)%, (62.03 ±1.97)% and (73.93±1.10)%, respectively. While the inhibitory rates of DDP+ BC-MSCs group were (8.27±0.63)%, (11.50±1.30)%, (20.57±0.93)%, (32.60 ±1.90)%, (52.27±0.73)% and (62.13±2.17)%, respectively. The inhibitory rates of DDP+ BN-MSCs group were (12.90±1.60)%, (16.53±2.87)%, (25.90±1.50)%, (39.40±2.40)%, (57.40±0.70)% and (69.03±1.07)%, respectively. The inhibitory rates of DDP+ BC-MSCs group were significantly lower than those of DDP group (P<0.05). The apoptotic rates of MCF-7 cells in DDP group, DDP+ BC-MSCs group and DDP+ BN-MSCs group were (47.77±1.98)%, (29.20±2.12)% and (37.92±2.21)%, respectively. The apoptotic rates of DDP group was significantly higher than that of DDP+ BC-MSCs group (P<0.05). The cell viabilities of MCF-7 in DDP group, DDP+ BC-MSCs group and DDP+ BN-MSCs group were 0.52±0.02, 0.72±0.02 and 0.64±0.02, respectively. The cell viability of DDP group was significantly lower than that of DDP+ BC-MSCs group (P<0.05). The result of Luminex liquid chip analysis showed that, the level of IL-6 in BC-MSCs group increased 2.50±0.68 fold when compared with BN-MSCs group (P<0.05). The relative expressions of IL-6 mRNA in DDP group and DDP+ BC-MSCs group were 1.02±0.10 and 7.58±0.55, respectively, with a statistically significant difference (P<0.01). The apoptotic rates of MCF-7 cells in DDP+ BC-MSCs group with or without IL-6 neutralizing antibody were (27.41±1.95)% and (42.45±2.87)%, respectively, with a statistically significant difference (P<0.05). The cell viabilities of MCF-7 cells in DDP+ BC-MSCs group with or without IL-6 neutralizing antibody were (72.40±2.60)% and (59.76±3.89)%, respectively, with a statistically significant difference (P<0.05).@*Conclusions@#BC-MSCs and BN-MSCs have been isolated and cultured successfully. Compared with BN-MSCs, BC-MSCs could attenuate the effect of DDP on MCF-7 cells, evidently decrease the apoptosis and increase the proliferation and vitality in an IL-6 dependent manner.
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Objective To investigate the expressions of TK1 (thymidine kinase 1) in PHC (primary hepatic carcinoma). Methods TK1 and AFP in serum of 33 cases of PHC (primary hepatic carcinoma), 38 cases of hepatic cirrhosis,36 cases of hepatitis and 35 cases of healthy people were detected by means of Western blot—enhanced chemiluminecence and electrochemiluminescence. Results The difference of TK1 level in PHC group indicated significance when compared with that in hepatic cirrhosis group , hepatitis group and control group (U value was 436.4, 352.1, 163.6, respectively, all P 0.05). Kaplan-Meier curve analysis revealed that PHC patients with TK1≤ 2.0 pmol/L had a significantly shortened overall survival when compared with those with TK1 > 2.0 pmol/L (χ2 = 3.954,P < 0.05). Multivariable Cox regression analysis indicated that the level of TK1 and TNM stage were the independent risk factors for patients with PHC (all P <0.05). Conclusions The detection of TK1 has a certain clinical value in the diagnosis, monitoring and evaluation of the prognosis of the PHC.
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Objective To investigate the expression of helper T cells (Th)17/regulatory T cells (Treg) balance associated factors in rheumatoid arthritis (RA) patients and their correlation with serum midkine (MK).Methods A total of 60 RA patients were divided into active RA patients (n=32) and inactive RA patients group (n=28).MK level in sera was detected by enzyme linked immunosorbent assay (ELISA) in 60 patients with RA and 30 healthy controls (HCs).The fraction of CD4+CD25+FOXP3+ Treg cells and IL-17+CD4+ Th17 cells in RA patients and healthy controls were determined by flow cytometry (FCM), and the expreasion of Foxp3, RORγt, Signal transducer and activator of transcription (STAT) 3 and STAT5 mRNA were detected with real-time polymerase chain reaction (PCR).Results were evaluated using ANOVA followed by q tests for comparisons of Th17 population between active RA patients, inactive RA patients and HCs, t test was used for comparing of Foxp3, RORγt, STAT3, STAT5 mRNA between RA group and HCs.The correlations between serum MK concentration and peripheral Treg cells, Th17 cells, Foxp3, RORγt, STAT3, STAT5 mRNA were analyzed by Pearson's correlation analysis.Results The percentages of Treg cells from active RA patients, inactive RA patients and HCs were significantly different (F=129.6, P＜0.01), the percentages of Treg cells of active RA patients [(1.41±1.05)％] were lower than that of the inactive RA patients [(3.6±1.6)％;q =7.92, P＜0.05] and healthy group [(7.7±1.7)％;q=22.45, P＜0.05], and there was significant difference between healthy group and inactive RA group (q=14.53, P＜0.05).The percentages of Th17 cells of the three groups were also significantly different (F=36.3, P＜0.01),the percentage of Th17 cells of active RA patients [(1.84±1.01)％] was significantly higher than that of inactive RA patients [(0.71±0.28)％;q=9.59, P＜0.05] and healthy group (0.53±0.16)％ [(P＜0.05;q=1 1.10, P＜0.05], there was no significant difference between the inactive RA group and healthy group (q=1.51, P＞0.05).The expression of RORγt and STAT3 mRNA in RA patients was higher than that of healthy controls (t=5.84, P＜0.01;t=4.52, P＜0.01).The expression of Foxp3 and STAT5 mRNA in RA patients were lower than healthy controls (t=6.01, P＜0.01;t=2.18, P＜0.05).Serum MK values were correlated with STAT5 (r=-0.55, P＜0.01), but not with Foxp3, RORγt, STAT3 mRNA or the percentage of Treg/Th17 cells.Conclusion Serum MK expression and the percentage of Th17 cells increase, while the percentage of Treg cells decrease in RA patients.Serum MK values are negatively correlated with STAT5 mRNA which is associated with Th17/Treg balance.This may be important in the pathogenesis of RA.
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Objective To evaluate the detection of membrane neutrophilic alkaline phosphatase ( mNAP) by flow cytometry in diagnosis of bloodstream infection .Methods A total of 298 patients with suspected bloodstream infections admitted in the First People ’ s Hospital of Lianyungang during June 2013 and October 2014 were enrolled;80 healthy subjects in physical examination center were also enrolled as the control group.Bloodstream infection was diagnosed by blood culture and mNAP was detected by flow cytometry.Serum levels of procalcitonin (PCT) and C-reactive protein (CRP) were detected by electro-chemiluminescence (ECL) and immune scatter turbidimetry , respectively.The value of mNAP, PCT and CRP in diagnosing bloodstream infection was determined by receiver operating characteristic ( ROC) curve. Results Among 298 patients, 109 were confirmed with bloodstream infections , including 43 patients with Gram-positive bacterial infections and 66 with Gram-negative bacterial infections .The median levels of CRP , PCT and mNAP in bloodstream infection group were 138.71 mg/L, 7.04 ng/mL and 13 929 AB/c, which were significantly higher than those in healthy control group (1.50 mg/L, 0.12 ng/mL, 1 831 AB/c;U=5.00, 48.50 and 65.01, P<0.01).The expression of mNAP in Gram-positive bacterial infection group was 9 598 ( 6 064-11 643 ) AB/c, which was significantly lower than that in Gram-negative bacterial infection group [16 512 (11 654-22 001) AB/c] (U=250.00, P<0.01).ROC curve analysis showed that, the areas under the curve (AUCs) of mNAP, PCT and CRP in diagnosing bloodstream infection were 0.987, 0.962 and 0.901.When 4 578AB/c, 0.90 ng/mL and 13.50mg/L were taken as optimal cut-off values, the sensitivities of mNAP, PCT and CRP in diagnosis of bloodstream infection were 95.8%, 93.0%and 90.3%; the specificities were 97.8%, 95.6% and 85.5%, respectively.Conclusion Among mNAP, PCT and CRP, mNAP is of the highest value in diagnosing bloodstream infection , and may be used as a biomarker for clinical diagnosis of bloodstream infection .
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Background and purpose:Recently, studies showed that non-steroidal anti-inlfammatory drugs (NSAIDs) could reduce the incidence of cancer. Whether ibuprofen could inhibit the growth of hepatocellular carcinoma cells had not been reported yet. In the current study, we investigated the effects of ibuprofen on hepatoma carcinoma BEL-7402 cells and the relevant mechanisms. Methods: Hepatocellular carcinoma BEL-7402 cells were randomly divided into 7 groups:the control group and the ibuprofen groups (0.1, 0.5, 1.0, 2.0, 3.0 and 4.0 mmol/L). The effect of ibu-profen on BEL-7402 HCC cells was measured by MTT method, the cell cycles were analyzed by flow cytometry (FCM), cell vitality and apoptosis were determined by cell analyzer. PCNA, Cyclin D1, Bcl-2 and COX-2 protein levels were examined by Western blot, and the expressions of prostaglandin E2 (PGE2) were measured by ELISA. Results:After the exposure to ibuprofen, the suppression ratio of BEL-7402 cells was increased (P<0.05). BEL-7402 cell vitality was decreased by degrees significantly (P<0.05), early apoptosis of BEL-7402 cells was increased (P<0.05), and the G0/Gl phase ratio was increased significantly compared with control group (P<0.05). Ibuprofen effectively decreased PCNA, Cyclin D1, Bcl-2 and COX-2 expressions in BEL-7402 cells (P<0.05), and decreased PGE2 protein expression in cell culture supernatants sig-nificantly (P<0.05). Conclusion:Ibuprofen is effective for inhibiting the proliferations, increasing apoptosis and blocking cell cycles of BEL-7402 HCC cells. The anti-tumor mechanisms of ibuprofen may be related with the inhibition of COX-2 and PGE2 expressions.
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The aim of this study was to develop a formulation to improve the oral absorption of baicalin (BA) by combining a phospholipid complex (PC) and self-emulsifying microemulsion drug delivery system (SMEDDS), termed BA-PC-SMEDDS. BA-PC was prepared by a solvent evaporation method and evaluated by complexation percentage (CP). The physicochemical properties of BA-PC were determined. The synergistic effect of PC and SMEDDS on permeation of BA was studied in vitro with Caco-2 cells and in situ with a single pass intestinal perfusion model. The improved bioavailability of BA in BA-PC-SMEDDS was confirmed in an in vivo rat model. The CP of BA-PC reached 100% when the molar ratio of drug to phospholipid (PP) was ≥1:1. The solubility of BA-PC increased in both water and octanol, and the log P o/w of BA-PC was increased significantly. BA-PC-SMEDDS could be dispersed more evenly in water, compared to BA and BA-PC. Both the Caco-2 cell uptake and single-pass intestinal perfusion models illustrated that transport of BA in BA-PC was lower than that of free BA, while improved significantly in BA-PC-SMEDDS. The relative bioavailability of BA-PC(1:2)-SMEDDS was 220.37%. The combination system of PC and SMEDDS had a synergistic effect on improving the oral absorption of BA.
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Objective To establish a flow cytometric method for the detection of alkaline phosphatase (ALP) expression on the membrane of neutrophils (mNAP).Methods EDTA-K2 anticoagulant venous bloods were collected.Expression of mNAP in peripheral blood was measured by flow cytometry using a phycoerythrin (PE)-labeled anti-ALP monoclonal antibody.BD QuantiBRITE PE was used to generate a calibration curve for PE fluorescence and ratios of PE to anti-ALP antibody and detect the bound ALP antibodies per cell (antibodies bound per cell,AB/c).Preanalytical handling including anticoagulants (EDTA-K2,citrate,and heparin),storage temperature,storage time,and plasma ALP were optimized and measured the precision.The expression levels of mNAP from 481 healthy controls were measured to establish a clinical reference range.The mNAP levels of 84 patients with severe infection and local infection,39 patients with virus infection were determined by this method.Results For preanalytical handling,application of PBS washing can effectively eliminate the interference of plasma ALP.The mNAP levels were not influenced by different anticoagulants and storage conditions (stored for 12 h either at room temperature or 4 ℃).This method had preferable reproducibility (CV in batch were 2.01％-3.33％,average 2.67％ ; CV between batch were 5.80％-6.00％,average 5.90％).The median (quartiles) of mNAP in health controls were 1 758 (1 378-2 310) AB / c for men and 1 897 (1 369-3 249) AB / c for women.There was no significant difference between genders (U =27 140,P =0.243 8).The clinical reference ranges (2.5 percentile to 97.5 percentile) of mALP was 920.5-3 493.0AB / c.The expression levels of mNAP of patients with bacterial infections (13 532,9 756-16 869 AB / c) were significantly higher than those of patients with virus infection(1 143,536-2 012 AB / c) and healthy controls (1879,1399-2497 AB / c) (H=221.5,P＜0.01).Conclusion BD QuantiBRITE PE kit can be used to standardize flow cytometer settings and quantitatively detect molecules per cell.The flow cytometric method for detection of mNAP has important clinical application for differentiating bacterial and virus infection.
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Objective To establish a method for detecting the concentration of paraquat (PQ) and creatinine(Cr) in urine simultaneously by capillary electrophoresis.Methods Experimental methodological study.8 acute PQ poisoned patients who were treated in the First People's Hospital of Lianyungang from January 2011 to June 2012 were collected.2 were male,and 6 were female.The separation were carried out using a 25 mmol/L pH1.97 glycine-HCl buffer(containing 40 mmol/L NaCl) in a fused-sillica capillary tube of 47 cm ×75μm I.D.by capillary zone electrophoresis.Urine had been injected by pressure for 4 s after samples were centrifuged and diluted for 10 times with H2O.The detection were monitored by a diode-array detector at 200 nm while samples were separated at a voltage of 20 kV.A systemic methodological evaluation of this method was carried out (The linear range,detection limit,repeatability test,recovery test and interference test).And the method was used to detect the concentration of PQ and Cr in PQ poisoned patients' urine.Results The peaks of PQ and Cr appeared within 5 min.The linear ranges of PQ and Cr were 2-1000,10-5000 μmol/L,respectively,with the correlation coefficients of 0.9997 and 0.9999 (P ＜0.01).The detection limits were 1.0 μmol/L for PQ and 5.0 μmol/L for Cr.The mean within-day(n =10) CVs of peak area for PQ and Cr were 2.84％ and 1.72％,while the mean inter-day(n =10) CVs of peak area were 3.62％ and 3.06％.The average recovery rate of PQ and Cr were 88.6％ and 90.2％ respectively.Diquat(DQ) didn't interfere with the assay.The range of PQ/Cr(μmol/mmol) for 8 cases was 8.9-2215.Conclusions A method was established successfully for the rapid determination of PQ and Cr in urine by capillary electrophoresis.The CE method devised here for direct measurement of urinary PQ and Cr from PQ poisoned patients is simple,fast,automatic and with good repeatability.It is an ideal method for rapid detection of urinary PQ in PQ poisoned patients.
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Objective To investigate the relation between the apotosis of B cells in the peripheral blood (PB) and the expression of interleukin (IL)-17 in patients with rheumatoid arthritis (RA).Methods The proportions of apoptosis of B cells in the PB of 80 patients with RA and 80 healthy controls were measured by flow cytometry.B cells in the PB of 20 RA and 20 healthy individuals were isolated by MACS and Western blotting was used to detect the Bcl-2 and Caspase-3 protein levels.IL-17 levels were detected by enzyme-linked immunosorbent assay (ELISA).T-test and linear regression were used to analyze the data.Results The proportions of apoptosis of B cells in the PB of patients with RA and healthy controls were (14±6)％ and (24±9)％ respectively.The rate of apoptosis of B cells in patients with RA was significantly less than healthy controls (t=2.737,P=0.021).The Bcl-2 protein level of B cells in the PB of patients with RA group was significantly higher than that of control group (26±10,12±6,P＜0.01).Conversely,the Caspase-3 protein level of B cells in the PB of patients with RA group was significantly lower than that of the control group (16±7,31±12,P＜0.01).ELISA detected elevated level of serum IL-17 in the patients with RA as compared with controls [(69±19),(27±10) pg/ml,t=4.631,P=0.014].There was a negative correlation between the level of IL-17 and apoptosis of B cells in patients with RA (r=0.36,P＜0.01).Conclusion The elevated bcl-2 and reduced caspase-3 of B cells in patients with RA further proves there is abnormal apoptosis of B cells in RA patients.There is negative correlation between the expression of IL-17 and apoptosis of B cells in patients with RA and IL-17 can inhibit B cell apoptosis.
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Objective To investigate the clinical significance of apoptosis of B cells in the peripheral blood of patients with rheumatoid arthritis (RA).Methods The proportions of B cells in the peripheral blood (PB) of 51 active and 30 remission patients with RA and 80 healthy controls were detected by flow cytometry.B cells in the PB of 10 active,10 remission and 10 healthy individuals were isolated by MACS.The apoptosis of cultured B cells,which were collected at 24,48,72,96 h respectively,were assessed by flow cytometry.ANOVA,t test and,Spearman correlation analysis were used for statistical analysis.Results The proportions of B cells marked as CD19 and CD22 in the PB of active and remission patients with RA and healthy controls were (26±11)％,(12±8)％,(10±7)％,(26±10)％,(12±8)％,(11±5)％ respectively.The proportions of B cells in the PB of active patients was significantly higher than that of remission patients and healthy controls (P＜0.01).There was positive correlation between B cell proportion in the PB of active patients and DAS 28 and IgG level.The proportion of apoptosis of B cells in the PB with active patients was less than healthy controls.Conclusion The pathway of apoptosis of B cells in the PB of active patients is inhibited,which could increase B cell proportion.Moreover,the high proportion of B cells in the PB of active patients is closely related to disease activity.
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Objective To study the potential use of the urinary beta-trace protein ( βTP) for diagnosis of type 2 diabetic renal injury.Methods 174 patients with type 2 diabetic mellitus (T2DM) were classified into 2 groups according to the ratio of urinary albumin to creatinine (Alb/Cr):diabetes without renal injury group (group A) and diabetes with renal injury group (group B).70 healthy subjects served as normal control group ( group C).The level of urinary βTP and αl microglobulin (α1MG) was measured by latex particle enhanced immunoturbidimetry assay.The urinary Alb and Cr were determined by nephelometry and Jaffe method respectively.The level of uriuary βTP among all groups was compared and ROC curve analysis was performed.The relevant analysis on urinary βTP,urinary α1MG and other related indexes was made.Results The median level of urinary βTP/Cr in group B was 9.1mg/g Cr,significantly higher than 3.1mg/g Cr of group A and 2.0mg/g Cr of group C.The difference had statistical significance ( H =45.5,P ＜ 0.01).The other indexes ( Alb/Cr,α1MG/Cr,SCr) were all higher in group B than in the other 2 groups ( H =110.9,38.3,11.4 respectively,P ＜0.01).The relevant analysis showed that urinary βTP/Cr was positively correlated with urinary α1MG/Cr (r =0.894,P ＜ 0.01),SCr (r =0.367,P ＜ 0.05 ),HbA(J) C ( r =0.242,P ＜ 0.05 ),systolic pressure ( r =0.162,P ＜0.05 ),and the course of the disease ( r =0.251,P ＜ 0.05 ).No correlation was found between urinary βTP/Cr and diastolic pressure,fasting blood glucose(FBG) or BMI.ROC curve analysis showed the area under the curve (AUC) was 0.86 (95％CI,0.78-0.93)for urinary βTP/Cr and 0.76 (95％ CI,0.67-0.85) for urinary α1MG/Cr.The best cut-off value of urinary βTP/Cr and α1MG/Cr was 4.1mg/g Cr vs 10.9mg/g Cr,the sensitivity was 68.5％ vs 59.7％,and the specificity was 89.8％ vs 80.3％.The difference had statistical significance (P ＜ 0.05).Conclusions Urinary βTP has better diagnostic value for type 2 diabetic patients with renal injury than urinary α1MG.It can sensitively reflect renal tubular injury and can be used as a novel available biomarker to evaluate the renal tubular injury in clinic.
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Objective To establish a method for detecting urinary vanillylmandelic acid (VMA), homovanillic acid (HVA) and creatinine (Cr) simultaneously by high performance capillary electrophoresis (HPCE). Methods The separations were carried out using a 120 mmol/L phosphate buffer (pH 6.80) in a fused-silica capillary tube of 47 cm×75 μm I.D. by capillary zone electrophoresis (CZE). Injections were made by using the pressure mode for 4 s at 1 p. s. i. after samples were centrifuged and diluted. The detections were monitored by a diode-array detector (DAD) at 200 nm after samples were separated at a voltage of 20 kV. The method developed was validated systematically and applied to urine samples from healthy adults (n = 100) and children (n = 100) for establishing the reference ranges of VMA/Cr and HVA/Cr, respectively. Results Under these conditions, the separations of VMA, HVA and Cr could be completed within 13 min. The linearity ranges of VMA, HVA and Cr were 0-500, 0-500 and 0-4 000 μmol/L, respectively, with the correlation coefficients (r) between 0.997 2 and 0. 999 1 (P < 0.01). The detection limits (S/N= 3) were 1.0 μmol/L for VMA, 1.0 μmol/L for HVA and 50.0 μmol/L for Cr. The mean within-run (n = 10) CVs of migration time for VMA, HVA and Cr in urine were 0.58%, 0.56% and 0.25% respectively, while the mean between-run (n = 10) CVs of migration time were 0.95%, 1.00% and 0.48% respectively. The mean within-run (n = 10) CVs of peak area for VMA, HVA and Cr were 3.78%, 3.97% and 2.76% respectively, while the mean between-rim (n = 10) CVs of peak area were 4.60%, 4.08% and 4.42% respectively. The average recoveries were 98.36% for VMA, 93.56% for HVA and 98.85% for Cr. Other compounds in human urine such as catecholamines, 5-hydroxytryptamine and albumen didn't interfere with the assay. The correlation between CE method and HPLC method was good. And the correlation coefficients (r) of VMA and HVA were 0.954 9(P <0.01) and 0.945 1 (P < 0.01), respectively. Skewness distributions were presented for VMA/Cr and HVA/Cr in random urine from both adults and children, and the 95% reference ranges were established by the percentile method. For adults, the reference ranges of VMA/Cr and HVA/Cr were 0-4. 26 and 0-1.69 (μmol/mmol), respectively. For children, the reference ranges of VMA/Cr and HVA/Cr were 0-10.39 and 0-4.31 (μmol/mmol), respectively. Conclusions The CE method devised here for direct measurement of urinary VMA, HVA and Cr is simple, fast,precise and automatic with good repeatability. It is an ideal method for routine detection and mass screening of pheochromocytoma and neuroblastoms.
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Objective To investigate the expression of serum sHLA-G in systemic lupus erythematosus (SLE) patients and the association with the disease activity.Methods The serum concentration of sHLA-G in SLE patients and healthy controls was measured with enzyme-linked immunosorbent assay.Results Significant higher sHLA-G levels were detected in patients of SLE than control group (P＜0.01),The serum concentrations of sHLA-G in active SLE patients were markedly higher than stable SLE patients (P＜0.01).The expression level of sHLA-G showed positive correlations with SLE activity index (SLEDAI)(r=0.30,P=0.01).There was no correlation between sHLA-G levels and serum concentration of Anti-dsDNA,C3,C4 and Anti-ANA in SLE patients (P＞0.05).Conclusion The level of serum sHLA-G is significantly increased in SLE patients.Positive correlations are observed between sHLA-G levels and SLEDAI.These data indicated that sHLA-G may play certain roles in the pathogenesis and progress in SLE.
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Objective To establish a rapid and sensitive method for detection of urinary protein.Methods In B-R buffer solution with pH 4.2,the signals of resonance light scattering by Poncesu S (PS) combined with protein in ?ex=?em=306nm were detected.Results There was a linear relation between the scattering signals of resonance light,and the protein concentration ranged from 0 to-1500 mg/l. The regression equation was ?I=2.24c-0.41,r=0.999 and the detection limit was 1.48 mg/l. The average recovery was 102.8% and the between-and within-subject coefficients of variation were 2.09% and 5.40% respectively.No significant difference was found compared with the method of PS.Conclusion The established method in this study is a simple,rapid and high sensitive method for determination of urinary protein.