Objective To investigate the relationship between levels of serum matrix metalloproteinases(MMPs)and brain edema and neurologic impairment in patients with intracerebral hemorrhage(ICH).Methods The levels of serum MMP-9 and MMP-2 in 31 patients with ICH were tested with ELISA for at 1 d,3 d,7 d and 2 weeks after onset.The volumes of hematoma and its peripheral edema were evaluated by CT,the neurologic impairment was evaluated by NIHSS at 1 d and 14 d after onset.Results Levels of serum MMP-9 and MMP-2 were significant higher in ICH group at each time point after onset than those in normal control group(allP

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.

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.

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.

Objective To investigate the diagnostic values of serum free fatty acids (FFA) and urine podocalyxin (PCX) in the patients with type 2 diabetic nephropathy(DN).Methods A total of 120 patients with type 2 diabetes mellitus (DM) were enrolled,and they were divided into three groups,normal albumin group(NA),microalbuminuria group(MA) and heavy albuminuria group(HA),based on the urinary albumin-creatinine ratio(UACR).In addition,40 healthy persons participated in medical examination were selected as the control group.Serum FFA levels were detected by a biochemical analyzer and urine PCX levels by ELISA.The correlation between them was analyzed by Pearson correlation,and their diagnostic values in type 2 DN were evaluated by the receiver operating characteristic curve (ROC).Results Serum FFA levels in the NA,MA,HA and control groups were (0.61 ± 0.14),(0.81 ± 0.13),(0.95 ± 0.18) and (0.49 ± 0.11) mmol/L,respectively,and urine PCX levels were (1.86 ± 0.45),(4.47 ± 1.48),(6.72 ± 1.40)and(1.38 ±0.24) ng/mL,respectively.The levels of serum FFA and urine PCX in type 2 DM patients were significantly higher than those in the control group(P ＜ 0.05),and increased with the progression of type 2 DM.Pearson correlation analysis showed that serum FFA levels were positively correlated with urine PCX levels(r =0.73,P ＜ 0.05).The sensitivity of serum FFA,urine PCX and combined detection in the diagnosis of type 2 DN were 74.1％,80.6％ and 89.5％,respectively,and the latter significantly higher than the former two (P ＜ 0.05).Conclusion Serum FFA and urine PCX levels increase significantly in diabetic patients with renal injury,and both of them have important clinical values in the diagnosis of type 2 DN.

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.

Objective Analysis of the expression of CD38,CD133 antigen and their clinical significance in myelodysplastic syndrome (MDS).Methods CD38 and CD133 antigen were analyzed by flow cytometry in 31 cases of MDS patients.Results CD38 was expressed in 18 cases (58.1% ),among them,12 cases were found to be myelodysplastic syndrome refractoryanermia ( MDS-RA ),accounting for 57.1%,6 cases were found to be MDS-RAEB,accounting for 66.7%.CD133 was expressed in 20 cases(64.5% ) ,among them,11 cases were found to be MDS-RA ( 52.4% ),1 case MDS-RAS,and 8 cases of MDS-RAEB,accounting for 88.9% .CD38 expressed significantly higher in MDS than anemia and relatively normal group ( P ＜ 0.05 ).CD133 expression in anemia groups was different from MDS-RA without statistical significance ( P ＞ 0.05 ),but was significantly different from relatively normal group (P ＜0.05).CD133 expression was significantly higher in these with MDS-RAEB than those in anemia and normal group ( P ＜ 0.05 ).Conclusions Combining with conventional antibodies,flow cytometry used in detection of CD38 ,CD133 ,could improve the diagnostic rate of MDS.

Objective To analyze and identify the chemical constituents of essential oil from the fruits of Cinnamomum camphora chvar. Borneol. Methods The essential oil from the fruits was extracted by steam distillation and its chemical constituents were analyzed and identified by GC-MS. Results Fifty compounds were separated, and 42 kinds of which accounting for 99.536% were identified. D-borneol was the most abundant compound, of which the amount was 50.684%of the total constituents. Conciusion This present study demonstrated higher content of natural D-borneol, providing scientific basis for further exploration and utilization of the fruits of Cinnamomum camphora chvar. Borneol.

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.

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.