Objective: To investigate the detection ability of two reagents by automated coagulation analyzer ACL-TOP using immune turbidimetric method, and to determine the merits and demerits of them. Methods:Two reagents were compared in the study. We selected 532 cases of hospitalized patients in Anzhen Hospital, 388 cases of which were unstable angina, 60 cases were pulmonary thromboembolism, 70 cases were coronary atherosclerosis, 14 cases were abdominal aneurysm, and the rest were the patients with high blood pressure and coronary heart disease. The plasmas of all the patients were extracted, and D-dimer was detected on ACL-TOP by D-Dimer (referred to as reagent A) and D-DHS (referred to as reagent B). Results:The results of D-dimer increased in different degrees, and there are 88 cases of false positive cases. The case whose FDP and individual D-dimer of D-dimer simultaneous detection was elevating and clinical symptoms were disappeared was detected by the VIDAS instrument platform verification. There were significant differences between A and B. The detection result of reagent A was [1594.15±2286.18], and reagent B was [912.97±1814.06]. There were significant differences between them (t=3.654, P<0.05). In addition, the consistency of the two reagents was poor (r=0.419, P<0.05). Conclusion:Comparing two reagents to detect D-dimer quantitatively by automated coagulation analyzer ACL-TOP, D-DHS owns greater ability to exclude false-positive results, so does the ability of anti-interference with jaundice, hemolysis and chylemia.

Objective To observe the function and survival of parathyroid cells cultured under simulated microgravity condition after cotransplanation of syngeneic allogeneic testicular sertoli cells.Methods Parathyroid cells in SD rats were assigned to flask-culture or bioreactor-culture.Allogeneic testicular sertoli cells in SD rats were cultured by using routine method.The recipients of hypoparathyroidism Wistar rat models were divided into 3 groups randomly:group A,receiving parathyroid cells(cultured with routine method)transplantation only;group B,receiving parathyroid cells and allogeneic testicular sertoli cells(cultured with routine method)transplantation;group C,receiving parathyroid cells(cultured under simulated microgravity condition)and allogeneic testicular sertoli cells transplantation.Allograft survival,change in cell components,apoptosis of infiltrative lymphocytes and parathyroid cells function were analyzed after transplantation respectively.Results The average survival time in group A,B and C was(17.3±1.6),(43.2±2.4)and (52.5±1.5)days,respectively.There was significant difference among group (P<0.05).In group C,parathyroid cells with strong secreting function grew into scaffold materials and adhered to them.FasL-expressing testicular cells and apoptotic lymphocytes were quite evident between allograft and kidney parenchyma.Conclusion Parathyroid cell cultured under simulated microgravity condition enhances its survival and function after cotransplanation of allogeneic testicular sertoli cell with immune privilege.

Objective To investigate whether B-cell activating factor (BAFF) involved in the patho-genesis of lupus nephritis (LN) by regulating phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling. Methods Twenty-eight lupus nephritis patients and 20 controls were included in this study. The clinical data were collected. BAFF levels in plasma were measured by ELISA, and the relationship between systemic lupus erythematosus disease activity index (SLEDAI) and BAFF were analyzed. The mRNA and protein levels of BAFF, phosphorylated-PI3K (p-PI3K), phosphorylated-Akt (p-Akt), phosphorylated-mTOR (p-mTOR) and Bcl-2 in kidney tissues were measured using real-time polymerase chain reaction (RT-PCR) and Western blotting. Data were analyzed using Mann-Whitney U test and Spearman correlation analysis. Results ①Plasma BAFF levels were significantly higher in LN patients [(580 ±45) ng/L] compared with controls [(208 ±30) ng/L](Z=-5.856, P<0.01), and significant positive correlation was found between plasma BAFF levels with SLEDAI (r=0.723, P<0.01). ② Plasma BAFF level in LN patients was positively correlated with 24 h UP and anti-dsDNA titers (r=0.381, 0.461, P<0.05). The protein level of BAFF in kidney tissues was positively correlated with 24 h UP and anti-dsDNA titer (r=0.469, 0.489, P<0.05).③The mRNA levels of BAFF, p-PI3K, p-Akt, p-mTOR and Bcl-2 in kidney tissues were increased in patients compared to controls[5.8±1.8 vs 2.1±0.7, Z=-4.915, P<0.01;6.7±0.9 vs 1.71±0.53, Z=-5.857, P<0.01;5.6±0.9 vs 1.8 ±0.5, Z=-5.751, P<0.01; 5.6 ±1.4 vs 1.6 ±0.4, Z=-5.291, P<0.01; 2.11 ±0.36 vs 1.33 ±0.22, Z=-4.844, P<0.01].④The protein levels of BAFF, p-PI3K, p-Akt, p-mTOR and Bcl-2 in kidney tissues were increased in patients compared to controls [0.72±0.19 vs 0.31±0.05, Z=-4.747, P<0.01;0.73±0.11 vs 0.33±0.09, Z=-5.834, P<0.01;0.77±0.06 vs 0.22±0.07, Z=-5.855, P<0.01;1.18±0.27 vs 0.47±0.13, Z=-5.416, P<0.01;2.08±0.37 vs 1.32±0.18, Z=-4.998, P<0.01]. Conclusion The findings of this study indicate that BAFF may participate in the pathogenesis of LN by regulating PI3K/Akt/mTOR signaling.

OBJECTIVE:To determine the contents of salicylic acid and the related substances in salicylic acid ointment by HPLC.METHODS:The determination was performed on Kromasil ODS-C18 chromatographic column with mobile phase consisted of methanol-water(adjust pH=2.5 using acetic acid,50∶50)at a flow rate of 1.0 mL?min-1.The sample size was 20 ?L and the detection wavelength was set at 270 nm.RESULTS:The main peak and the related substances were well-separated.The linear range of salicylic acid was 10.014～100.14 ?g?mL-1(r=0.999 9)with an average recovery of 99.35%(RSD=0.99%)and a lowest detectable limit of 5 ng?mL-1,the labeled amout of related substances all less than 1.01%.CONCLUSION:The method developed in our study is simple in operation,accurate in results,specific and suitable for the content determination of salicylic acid ointment.

Objective To explore the circadian rhythm of interleukin (IL)-6 in collagen induced arthritis (CIA) rats and investigate the effectiveness and safety of methotrexate (MTX) administered according to IL-6 rhythm.Methods Serum IL-6 concentrations of CIA and normal rats at different time points were measured by enzyme-linked immunosorbent assay (ELISA).CIA rats were randomly divided into the experimental group A,experimental group B and control group.MTX (1/7 mg·kg-1 ·d-1) was administered once a day when the IL-6 level began to increase or decrease in the experimental group,while MTX (1 mg/kg) administered once a week in the control group.Arthritis scores and leukocyte counts were observed.The production of IL-6 and C reactive protein (CRP) levels in the serum were measured.Moreover,histological changes in the ankle joint were examined.One-way analysis of variance (ANOVA),two independent sample t test were used to evaluate the experimental data.Results The plasma IL-6 levels in CIA rats were different at different time points,which began to increase at 18:00 and decrease at 6:00.Arthritis score in the experimental group A (4.8±0.7)was lower than that in the control group (5.8±1.0,t=2.256,P=0.0406) and experimental group B (5.5±0.5,t=2.393,P=0.0313).The levels of TNF-α [(185±21) ng/L],IL-6 [(99±6) ng/L],IL-1β[(20.1±1.6) ng/L] and CRP[(1251±282) μg/L] in the experimental group A were significantly lower than those in control group [TNF-α:(207±10) ng/L,t=2.726,P=0.016 4;IL-6:(100±6) ng/L,t=2.669,P=0.0183;IL-1β:(22.0±1.3) ng/L,t=2.814,P=0.0138;CRP:(1 417±278) μg/L,t=2.369,P=0.0327].The level of IL-6 in the experimental group A[(93±6) ng/L] was lower than that in the experimental group B [(99±6) ng/L,t=2.323,P=0.0358].Compared with the control group [(9650±1062)/μl],the counts of leukocyte in the experimental group A [(4595±603)/μl,t=3.841,P=0.0064] and experimental group B [(3833±585)/μl,t=4.442,P=0.003] were significantly lower.Compared with the control group (9.1 ±2.0),histopathology scores in the experimental group A (6.6±1.3,t=2.606,P=0.015) and experimental group B (7.4±1.3,t=3.857,P=0.0007) were significantly lower.Conclusion The plasma IL-6 levels in CIA rats have shown evident circadian rhythm.There-fore,once a day administration is better than once a week.The therapeutic effect of RA may be improved by administering MTX at the time points according to the circadian rhythm of IL-6.

Objective To develop a HPLC method for the determination of chlorogenic acid in xiaoaiping injection in human plasma.Methods The analytical column was packed with Hypersil ODS C18 (250 mm×4.6 nm,5 μm).The mobile phase was acetonitrile-0.4% phosphoric acid solution(13:87).The flow rate was 1 ml/min,and detection wave length was set at 327 nm.Results The linear ranges were 2.5～60.0 mg/L chlorogenic acid in human plasma,with the correlation coefficients of 0.9996.The Low limit of detection was 0.25 μg/ml.The method and extraction recovery were 100.1%～102.6%and 78.6%～80.3%.The relative standard deviation value for the within-day and between-day precision were less than 3.9%.Conclusion The method is simple,sensitive for the determination of chlorogenic acid in xiaoaiping injection in human plasma.

Objective To investigate the relationship between the biological characteristics and the expres-sion of stromlysin (MMP-7) and microvessel density (MVD) in breast cancer tissue and the effect of MMP-7 on MVD of breast cancer. Methods 60 fresh samples were obtained from patients with breast cancer,and then the ex-pression of MMP-7 and vascular endothelial cell CD34 were studied by means of immunohistochemical assay (SP method). The relationship between MMP-7, MVD and age, tumor size, histological type, lymph node metastasis and expression of ER, PR, PCNA, p53 and CerbB2 was studied. The relationship between the biological characteristics and the expression of MMP-7 and MVD in breast cancer were evaluated. Results The positive expression rote of MMP-7 in the d>2 cm group,the group with lymph node metastasis,the group with positive expression of PCNA, p53 and C-erbB-2 (78.1%,74.2%,71.8%,67.6% and 72.2%)were all higher than that in the d≤2 cm group, the group without lymph node metastasis,the group without positive expression of PCNA,p53 and C-erbB-2(32.1%, 41.3%,38.1%,38.5% and 33.3%) (P<0.05). MVD was higher in the positive breast cancer tissue of the group of d>2 cm(34.61±6.97), the group with lymph node metastasis (34.37±7.50), PCNA ( 33.24±8.39), p53 (33.28±8.94), C-erbB-2 (33.55±8.57) than in the negative breast cancer tissue of the d≤2 cm group ( 28.60±9.82), the group without lymph node metastasis (27.48±8.66), PCNA (26.88±7.89), p53 (21.71± 7.59),C-erbB-2(27.42±27.69) (P<0.05). MVD(33.62±7.36)/ high power lens in the MMP-7 positive group was higher than that (27.86±9.45)/high power lens in the MMP-7 of negative group(P<0.05). The posi-tive expression of MMP-7 was correlated with MVD (r=0.380, P<0.05). Conclusion The high expression of MMP-7 and MVD is associated with the development and metastasis of breast cancer. MMP-7 could promote the mi-crovessel development in mammary cancer.

Objective:To Studythe extracellular matrix metalloproteinase inducer (CD147) changes in patients with systemic lupus erythematosus (SLE), and to study thecorrelation of CD147 level and athero-sclerosis in SLE.Methods:Eighty patients with SLE in total were divided into intimal thickening group, (24 cases, group A) and normal intimal group (56 cases, group B) according to carotid intimamedia thickness (IMT) checked by carotid ultrasonography. In addition, their age, bodymass index, blood pressure, seral total cholesterol (TC), high density liptein cholestero (HDL-C), low density lipoprotein cholestero (LDL-C), triglyceride (TG), urinary protein quantitative test(24 h), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), anticardiolipin antibody, serum creatinine level, course of the disease, and treatment regimens were collected. Thirty-five healthy people were set as the control group (group C). The levels of serum CD147 were measured by enzyme-linked immuno sorbent assay (ELISA) in 3 groups. The correlation between the serum CD147 level of SLE patients and atherosclerosis was analyzed. The statistical analysis was carried out with independent t-test, chi-square test, analysis of variance, Spearman correlation and Logistic regression. Results:① Levels of serum CD147 in group A [ (238±30) pg/ml] were significantly higher than those in group B [(198±30) pg/ml] and group C [(150±26) pg/ml, F=67.908, P<0.01]. ② Body mass index, hypertensive ratio,total blood cholesterol,urine protein quantitative test (24 h), systemic lupus erythematosus disease activity index (SLEDAI), serum CD147 index in group A were significantly higher than those in group B ( P<0.05). ③ In Logistic regression analysis, serum CD147 [ OR (95% CI)=1.039(1.014, 1.065), P<0.05], urine protein quantitative (24 h) [ OR (95% CI)=2.598(1.033, 6.534), P<0.05] were independently relevant factors affecting carotid artery IMT. Conclusion:Serum CD147 is an independent risk factor for carotid intimamedia thickness in SLE patients.

Objective In this study,maximum platelet aggregation rate of Light transmittance aggregometry (LTA) for coronary heart disease(CHD) patients taking antiplatelet drug and patients without antiplatelet therapy was measured in non-adjusted and platelet count-adjusted platelet-rich plasma (PRP).The aim of this study is to compare which method is superior in evaluation of antiplatelet drug effect.Methods This is a methodology comparative research.560 CHD outpatients and inpatients that visited Beijing Anzhen Hospital of the Capital University of Medical Sciences from May to June,2012 were chosen,who were treated with aspirin monotherapy,or patients on combination therapy with aspirin and clopidogrel,as well as patients without antiplatelet therapy.LTA was performed in non-adjusted (improved method) and platelet count (200 × 109/L)-adjusted PRP (original method),using 6 μmol/L adenosine diphosphate (ADP) and 0.5 mmol/L arachidonic acid (ARA) as agonists.The maximum aggregation rates in 5 min were detected,and consistency and differences of the two methods were compared.Results There is no statistically significant correlation between maximum aggregation rate and platelet count in PRP with 6 μmol/L ADP or 0.5 mmol/L ARA as agonists in all subgroups including aspirin monotherapy,combination therapy with aspirin and clopidogrel and patients without antiplatelet therapy (-0.21 ≤ r ≤0.111,P ＞ 0.05).The maximum aggregation rate using ADP as agonists in original method is decreased compared with improved method,there is statistically significant difference in all subgroups including patients without antiplatelet therapy,aspirin monotherapy,combination therapy with aspirin and clopidogrel less than one week and more than one week.The variability of platelet aggregation rate using ADP as agonists with improved method is lower than that with original method in all subgroups.Yet the maximum aggregation rates using ADP as agonists with improved method and original method correlate well with each other in all subgroups (r =0.78,0.73,0.40,0.71,P ＜0.01).In the subgroup of subjects without antiplatelet therapy using ARA as agonist,platelet aggregation rate is decreased in original method compared with improved method,there is statistically significant difference,and the variability of the aggregation rate with improved method is also lower than that with original method,ranging from 62％-98％ relative to 5％-89％.The decrease of aggregation rate using ARA as agonist for patients taking antiplatelet drug compared with patients without antiplatelet therapy can be detected both with improved method and original method.Conclusion Non-adjusted PRP in LTA is more convenient and time-saving,and it also means less effects on platelet in vitro.Therefore,non-adjusted PRP is more suitable for monitoring efficacy of antiplatelet therapy in clinical laboratory.

ObjectiveTo investigate the characteristics of trace elements and amino acid metabolism in the blood of patients with hepatocellular carcinoma (HCC) and to guide clinical treatment. MethodsThis study included 125 untreated patients with primary HCC in Tangshan Maternal and Child Health Hospital and Tangshan Guzhi District Hospital from March to November, 2013, as well as 50 healthy controls who underwent physical examination in Tangshan Maternal and Child Health Hospital. Blood trace elements were measured by atomic absorption spectrophotometry, and the levels of amino acids were determined by an amino acid analyzer. Comparison of continuous data was made by t test. ResultsCompared with the controls, the HCC patients had a significantly higher Cu level (t=6.30, P＜0.05) and significantly lower Zn, Fe, and Mg levels (t=10.15, 3.98, and 2.25, all P＜0.05). The HCC patients showed significant increases in tyrosine, phenylalanine, and methionine levels (t=10.10, 13.46, and 2.34, all P＜0.05) and significant decreases in valine, isoleucine, leucine, and arginine levels (t=3.65, 2.98, 5.67, and 8.35, all P＜0.05). ConclusionHCC patients show changes in various amino acids and trace elements in blood. Measurement of trace elements and amino acids is of certain significance for evaluating HCC patients′ condition and guiding clinical treatment.