The increased serum Th17 related cytokine concentration in patients with cryptococcal meningitis and its clinical significance / 国际检验医学杂志

Objective To investigate the IL‐17A/F and IL‐22 ,the Th17 related cytokine ,in the serum of cryptococcal meningitis patients and its clinical implications .Methods We collected peripheral blood from 22 cryptococcal meningitis patients and 20 matched healthy controls from February 2011 to February 2013 .Then the peripheral blood mononuclear cells(PBMC)were seperat‐ed with isodensity centrifugation .The RT‐PCR and ELISA were used to detect the mRNA and protein lever of Th17 related cyto‐kine such as IL‐17A ,IL‐17F and IL‐22 .In addition ,the relationship between these cytokines and cerebrospinal fluid characteristics were analyzed .Results The serum concentration of IL‐17A and IL‐22 were much higher than that of healthy controls (P<0 .01) . Their concentration were even higher in patients with poor prognosis factors ,including higher cerebrospinal fluid protein level and cryptococcal antigen titer ,lower cerebrospinal fluid glucose concentration and increased cerebrospinal fluid open pressure .Conclusion IL‐17A and IL‐22 were involved in the pathogenesis of cryptococcal meningitis and may be a potential prognosis biomarker for cryptococcal meningitis.

Diagnostic Value of Osteopontin for Ovarian Cancer:a Meta-Analysis / 现代检验医学杂志

Objective To evaluate the diagnostic performance of osteopontin (OPN)for ovarian cancer.Methods Wanfang, CQVIP and CNKI were retrieved to identify eligible studies on diagnostic value of OPN for ovarian cancer that published be-fore May,2014.The quality of the studies was evaluated by QUADAS tools.The diagnostic sensitivity,specificity,negative and positive likelihood ratios and diagnostic odds ratio (DOR)were pooled by random-effects models.The overall diagnostic performance was estimated by summary receiver operating characteristic (SROC)curves approaches.Results Six studies met the included criteria.The summary estimates for OPN in the diagnosis of ovarian cancer in the studies included were as follows:sensitivity 0.83 [(95% confidence interval(CI):0.78~0.87)],specificity 0.91 (95% CI,0.88~0.94),positive likelihood ratio 9.00 (95% CI,5.91~13.71),negative likelihood ratio 0.19 (95% CI,0.15~0.25),and DOR 47.58 (95%CI,27.93~81.05).The area under curve (AUC)for OPN was 0.87 with Q value of 0.80.Conclusion OPN has high diag-nostic value for ovarian cancer.

MiR-146a regulate the proliferation and interleukin-2 production of T helpers from primary biliary cirrhosis patients / 中华风湿病学杂志

Objective To investigate the effect of miR-146a on the proliferation and interleukin (IL)-2 production of T helper cells from primary biliary cirrhosis (PBC) patients.Methods MiR-146a in the peripheral blood mononuclear cells (PBMC),monocytes,T helper cells,cytotoxic T cells and B cells from 20 confirmede PBC patients and age/sex matched healthy controls were detected by quantitative PCR.By gainand-loss of function,the miR-146a's effect on anti-CD3/anti-CD28 activated T helper's proliferation and IL-2 production ability were measured by CCK-8 approach and enzyme linked immunosorbent assays (ELISA),respectively.Statistical analysis were carried out by t-test.Results PBMCs (0.46±0.20 vs 1.00±0.26; t=7.47,P＜0.01),T helpers (0.33±0.13 vs 1.00±0.14; t=6.15,P＜0.01) and monocytes (0.56±0.11 vs 1.00±0.11; t=4.97,P＜0.05),but not B cells (0.91±0.06 vs 1.00±0.14; t=0.97,P＞0.05) and cytotoxic T cells (0.98±0.15vs 1.00±0.12; t=0.22; P＞0.05) from PBC patients had lower miR-146a expression level than that of healthy controls.Inducible up expression of miR-146a was observed in PBC patients'T helpers stimulated with antiCD3/anti-CD28 (1.00±0.18 vs 9.12±2.05; t=8.81; P＜0.01).The activated T helpers from PBC patients had higher proliferative ability [PBC:0.35±0.06 (A); healthy controls:0.26±0.04 (A); t=2.83; P＜0.05] and increased IL-2 production [PBC: (685.60±109.19 pg/ml)]; Healthy controls: [(512.20±72.26) pg/ml; t=2.96; P＜0.05 ] than those of healthy controls.For activated T helpers,the proliferation ability,as well as IL-2 production,was enhanced by miR-146a.Conclusion MiR-146a can down regulate the proliferation and IL-2 releasing of activated T helpers.The reduced miR-146a expression enhances IL-2 production and promotes proliferation of T helper of PBC patients,thus,may be involved in the pathogenesis of PBC.

IL-22 is involved in atherosclerosis lesions by regulated impaired proliferation ability of oxidized low density lipoprotein treated CRL-1730 / 中华微生物学和免疫学杂志

Objective To investigate the association between IL-22 and the pathogenesis of coronary artery atherosclerosis(AS).Methods The relative expression of IL-22 mRNA in PBMC from 30 AS patients and 8 patients without any signs of coronary artery stenosis was detected by RT-PCR.Serum IL-22 levels of 22 patients without any signs of coronary artery stenosis and 79 AS patients were detected by ELISA.CRL-1730 cells(human umbilical vein endothelial cells) were stimulated with oxidized low density lipoprotein (ox-LDL) at different dosage for 24 h,and the expression of IL-22R1 was detected by flowcytometry.The proliferation ability of CRL-1730 cells treated with IL-22(20 ng/ml) and/or ox-LDL(100 μg/ml)was measured by MTS assay,and the expression of basic fibroblast growth factor(bFGF) was detected by RTPCR and ELISA.Results Decreased IL-22 expression in PBMC and serum was observed as worsen of AS.The expression of IL-22R1 in ox-LDL treated CRL-1730 cells was increased in dose dependent manner.OxLDL decreased proliferation ability,as well as bFGF expression and releasing,of CRL-1730 cells.This effect of ox-LDL was partially rescued by IL-22.Conclusion IL-22 may have anti-atherosclerosis effect.This effect may be mediated by regulating bFGF expression and endothelial cells proliferation ability in the presence of IL-22.

Increased expression of microRNA-146a in peripheral blood mononuclear cells of patients with chronic immune thrombocytopenic purpura and its clinical significance / 中华微生物学和免疫学杂志

Objective To investigate the increased expression of microRNA-146a(miR-146a) in peripheral blood mononuclear cells (PBMC) of patients with chronic immune thrombocytopenic purpura (ITP) and its clinical significance. Methods Twenty-eight patients with chronic ITP and 28 healthy controls matched with age and gender were enrolled in this study. Fluorescent quantitative PCR reaction was used to detect the relative expression of miR-146a in their PBMC. The serum concentration of TNF-α, IL-2,IL-1 β and IFN-γ were measured by ELISA. CCK-8 method was used to detect the proliferation ability of PBMC , which transfected with miR-146a mimics or inhibitor and then stimulated with platelet . Results The relative expression of miR-146a in ITP patients was higher than that of healthy controls. The increased expression of miR-146a was negatively correlated with the serum TNF-α, IL-2 and IFN-γ. The PBMC transfected with miR-146a mimics had reduced expression of IL-2 and proliferation when stimulated with platelet.In contrast, the opposite effect was observed with the miR-146a inhibitors transfection. Conclusion MiR146a was involved in the pathogenesis of chronic ITP by controlling IL-2 production and PBMC proliferation.Thus, it may be a potential therapy target for chronic ITP.

The role of IL-27 in patients with primary biliary cirrhosis and its clinical significance / 中华微生物学和免疫学杂志

Objective To detect the expression of IL-27 in PBC (primary biliary cirrhosis)patients and the possible involvement of IL-27 signal pathway in PBC. Methods The gene transcription and protein expression levels of IL-27 in patients with PBC, chronic hepatitis B(CHB) group and healthy controls(HC) were measured by real-time PCR, ELISA, flow cytometry and immunohistochemistry. AST,ALP, ALT, TBIL, GGT were determined and their correlation with IL-27 was also analyzed. Results IL-27 was significantly elevated in patients with PBC and IL-27 is present in the liver tissues of patients with PBC. Expression of IL-27 on CD4+T cells was increased in patients with PBC(72.40% ±6.22% ) and CHB(59.40% ± 7.03%) compared with HC(1.70%±0.55%,P＜0.01). Expression of IL-27 protein was increased in patients with PBC [( 126.25 ± 36.00 ) pg/ml] compared with CHB [( 51.81 ± 23.30 )pg/ml, P ＜ 0. 01] and HC[(34.19 ± 9.70) pg/ml, P ＜ 0.01], and it was positively correlated with GGT( r = 0.554, P＜0.01) and TBIL (r = 0.559,P＜0.01), but no correlation with ALT, AST, ALP.Conclusion These facts indicated the key role of IL-27 in the immune inflammatory reaction in patients with PBC.

Prediction of standard Gibbs energies of the transfer of peptide anions from aqueous solution to nitrobenzene based on support vector machine and the heuristic method.

Quantitative structure-property relationship (QSPR) method was performed for the prediction of the standard Gibbs energies (DeltaGtheta) of the transfer of peptide anions from aqueous solution to nitrobenzene. Descriptors calculated from the molecular structures alone were used to represent the characteristics of the peptides. The four molecular descriptors selected by the heuristic method (HM) in COmprehensive DEscriptors for Structural and Statistical Analysis (CODESSA) were used as inputs for support vector machine (SVM) and radial basis function neural networks (RNFNN). The results obtained by the novel machine learning technique, SVM, were compared with those obtained by HM and RBFNN. The root mean squared errors (RMS) of the training, predicted and overall data sets are 2.192, 2.541 and 2.267 unit (kJ/mol) for HM, 1.604, 2.478 and 1.817 unit (kJ/mol) for RBFNN and 1.5621, 2.364 and 1.756 unit (kJ/mol) for SVM, respectively. The prediction results were in agreement with the experimental values. This paper provided a potential method for predicting the physiochemical property (DeltaGtheta) of various small peptides.

Separation and determination of active components in Schisandra chinensis Baill. and its medicinal preparations by non-aqueous capillary electrophoresis.

A simple, economical and effective non-aqueous capillary electrophoresis separation and detection method was developed for the quantification of deoxyschizandrin and gamma-schizandrin in Schisandra chinensis Baill. and its medicinal preparations for the first time. After optimization of separation conditions, a buffer of 140 mmol/L sodium cholate in methanol was selected for separating the two analytes, but baseline separation of SA and SB in real samples was not obtained. Therefore second-order derivative electropherograms were applied for resolving overlapping peaks. Regression equations revealed good linear relationships (correlation coefficients 0.9975--0.9988) between peak heights in second-order derivative electropherograms and concentrations of the two analytes. The relative standard deviations (RSD) of the migration times and the peak height of the two constituents were in the ranges 0.62--0.79% and 0.25--2.17% (intra-day) and 1.43--2.06 and 4.08--5.72% (inter-day), respectively. The recoveries of the two constituents ranged from 93.2 to 103.0%. The results indicated that baseline separation of the analytes was sometimes hard to obtain in real samples and second-order derivative electropherograms were applicable for the resolution and analysis of overlapping peaks.

Rapid and sensitive determination of ephedrine and pseudoephedrine by micellar electrokinetic chromatography with on-line regenerating covalent coating.

A rapid, sensitive and reproducible micellar electrokinetic chromatographic method using hexamethyldisilazane as on-line regenerating covalent coating was developed for the quantification of ephedrine (E) and pseudoephedrine (PE). E and PE were derivatized with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazol for laser-induced fluorescence detection. The on-line regenerating covalent coating formed a combinative double coating with the subsequently produced dynamic SDS coating. The total coating can be easily removed and conveniently regenerated on-line. The simple coating procedure was described. By a series of optimization, a running buffer of 20 mm Na(2)B(4)O(7) + 16 mm SDS was applied for the separation of the derivatives. Linear relationships for E and PE were obtained in the range of 0.044-6.60 microg mL(-1) (correlation coefficients: 0.9975 for E, 0.9981 for PE), and the detection limits for E and PE were 1.71 and 0.67 ng mL(-1), respectively. The separation speed, the reproducibility and the sensitivity were much improved over those of other capillary electrophoresis methods more recently reported. The method was applied to the analysis of the two alkaloids in traditional herbal preparations with recoveries in the range 92.8-104.8%.

Separation and determination of two sesquiterpene lactones in Radix inulae and Liuwei Anxian San by microemulsion electrokinetic chromatography.

A novel microemulsion electrokinetic chromatography (MEEKC) method for separating and determining two sesquoterpene lactones, alantolactone (AL) and isoalantolactone (IAL), in Radix inulae and Liuwei Anxian San has been developed. The effects of several important factors such as internal organic phases, concentration of microemulsion, concentration of acetonitrile, injection time and running voltage were systematically investigated to determine the optimum conditions. The optimum microemulsion system was composed of n-hexane (0.32% w/w), SDS (1.24% w/w), 1-butanol (2.64% w/w), acetonitrile (10% w/w) and 10 mm sodium tetraborate buffer (85.80% w/w, pH 9.2). The applied voltage was 20 kV. The analytes were detected at 214 nm. Regression equations revealed linear relationships (correlation coefficients 0.9950 for AL and 0.9946 for IAL) between the peak area of each analyte and the concentration. The limits of detection (defined as a signal-to-noise ratio of about 3) were approximately 0.45 microg/mL for AL and 0.56 microg/mL for IAL. The levels of the analytes were successfully determined with recoveries ranging from 98.2 to 104.3%. Furthermore, a simple and effective extraction method, with methanol in an ultrasonic water bath for 60 min, was used for sample preparing. Also, MEEKC was compared with micellar electrokinetic chromatography (MEKC) and shown better separation results.

Microemulsion electrokinetic chromatography with laser-induced fluorescence detection: as tested with amino acid derivatives.

Over a decade ago, microemulsion electrokinetic chromatography was introduced as a novel mode of capillary electrophoresis. However, there has not been publication on the combination of microemulsion electrokinetic chromatography with laser-induced fluorescence detection. In this paper, a preliminary method using microemulsion eletrokinetic chromatography combined with laser-induced fluorescence detection and second derivative electrophoregram was established as a sensitive and selective assay for separation and determination of nine amino acids after derivatization with 4-chloro-7-nitrobenzo-2-oxa-1, 3-diazol. The derivatization and separation conditions were optimized. In the investigated concentration ranges correlation coefficients were better than 0.995. The relative standard deviation (n = 5) of the migration times and peak heights were 0.56-0.76 and 2.21-7.15%, respectively. The detection limits (S/N = 3) were at a neaomolar level (0.32-2.20 nM). The method was applied for the analysis of compound amino acid injection and a Chinese traditional herbal medicine. The recoveries were 95.9-107.9%.

Identification and determination of effective components in Euphrasia regelii by capillary zone electrophoresis.

A capillary zone electrophoresis (CZE) method has been developed for simultaneous determination of eukovoside, cinnamic acid and ferulic acid in Euphrasia regelii for the first time. The electrophoresis buffer was 20 mmol/L sodium borate containing 10% (v/v) methanol (pH 8.50). The effects of concentration of borate and electrolyte pH on electrophoretic behavior and separation were studied. Regression equations revealed linear relationships (correlation coefficients 0.9995-0.9998) between the peak area of each analyte and the concentration. The levels of analytes in the different parts of Euphrasia regelii were easily determined with recoveries ranging from 95.5 to 104.2%.

Determination of rhein, baicalin and berberine in traditional Chinese medicinal preparations by capillary electrophoresis with two-marker technique.

A CZE method for the identification and determination of three bioactive components, rhein, baicalin and berberine, was developed, with 10 mM borate at pH 9.20 as background electrolyte and direct UV detection at 254 nm. The two-marker (glycyrrhizin acid and cefalexin) technique was used to improve the repeatability of analysis. When the migration indices were used, the repeatability was greatly enhanced compared with migration times. The apparent dissociation contents of rhein and baicalin were also obtained. This method has been successfully applied to the simultaneous analysis of the three components with the recoveries from 96.7% to 104.6%.

Human serum albumen enhanced resonance light scattering of dyes.

For the first time, the relationship between the structural parameters of dye molecule and the enhanced resonance light scattering (RLS) of dye-human serum albumen (HSA) was investigated using chemometrics technique. The data set of the enhanced RLS intensity of the dye-HSA was obtained under the same experimental conditions for 22 dyes. After calculated with HyperChem software and selected by stepwise regression approach, six structural parameters for the maximum RLS wavelength were used in multivariable linear regression (MLR) (R=0.932), seven structural parameters for the corresponding enhanced RLS intensity were employed by MLR (R=0.934). According to these models, the maximum RLS wavelength and the enhanced RLS intensity are both attributed to the spatial structure and energies of dye.

Identification and determination of ecdysone and phenylpropanoid glucoside and flavonoids in Lamium maculatum by capillary zone electrophoresis.

The simultaneous determination of 20-hydroxy ecdysone (1), 3,7-dimethoxy-quercetin (2), acteoside (3) and rutin (4) in the mixture of leaf and stem, and the flower of Lamium maculatum has been investigated by capillary zone electrophoresis for the first time. With an electrolyte containing 30 mmol/L borate, at pH 9.47 and 20 kV applied voltage, the four active compounds were completely separated within 5 min with satisfactory results. The effects of concentration of borate and electrolyte pH on electrophoretic behavior and separation were studied. Regression equations revealed linear relationships (correlation coefficients 0.9998-0.9999) between the peak area of each analyte and the concentration. The levels of analytes in the different parts of Lamium maculatum were easily determined with recoveries ranging from 98.3 to 105.0%.

The fluorescence study of the inclusion coordinated compound of beta-cyclodextrin and p-hydroxyphenolacetamide and its analytical applications.

The inclusion coordinated compound fluorescence of p-hydroxyphenolacetamide (PHPA) with beta-cyclodextrin was studied and a high sensitive analytical method to determine the content of PHPA was established based on the increased fluorescence intensity of the coordinated compounds in Britton-Robinson buffer solution (pH 7). The wavelength of the excited emission are 282 and 310 nm, respectively. The enhanced coordinated compounds fluorescence intensity is proportional to the concentration of PHPA in the range 0.030-11.0 mg/l. The relative standard deviation (R.S.D.) were within 0.66-4.2%. Correlation coefficient and inclusion binding constant Kf are obtained within 0.9950-0.9996 and 4.67 x 10(2), respectively. Meanwhile, a linear increase of the resonance light scattering (RLS) with the inclusion coordinated compounds concentration was noticed at a synchronous wavelength of 584 nm. The forming reasons of inclusion coordinated compounds were investigated. The 1:1 molar ratio composition of inclusion compound was measured with DTA and elemental analysis. The method is simple, rapid and has a good reproducibility. The method was used for determination of PHPA in an artificial sample with satisfactory results.

Catalytic Spectrophotometric Determination of Osmium (Ⅳ) Using Osmium (Ⅳ)-KIO4-Chlorophosphonazo-mA System / 分析化学

A catalytic spectrophotometric method for the determination of micro amounts of osmium has been established based on the catalytic action of Os(Ⅳ) on the oxidation fading reaction of chlorophosphonazo-mA with KIO4 in alkaline-medium. The reaction conditions were optimized by orthogonal experimental design. The detection limit for osmium was 2.0 μg/L.Beer＇s law was obeyed in the range from 7.0 to 25.0 μg/L for Os(Ⅳ). The method has been applied to the determination of micro amounts of Os(Ⅳ) in concentrate of noble metals and secondary alloy,both of the relative errors were 0.9%. Recoveries varied from 95.38% to 106.0%.