Objective To investigate the number and activity changes of endothelial progenitor cells(EPCs) in hemodialysis patients,and explore its correlation with the risk factors of coronary heart disease.Methords Total mononuclear ceils(MNCs) were isolated from peripheral blood of patients with chronic renal failure in long-term hemodialysis and from a normal control group by Ficoll density gradient centrifugation and then were plated on humanfibronectin-coated dishes.Mter 7 days of culture,EPCs were characterized as adherent cells by double staining with FITC-UEA-I and DI-LDL,and were further identified by the expression of CD34,CD133 and KDR with flow cytometry.EPCs migration was determined with modified Boyden chamber assay.EPCs adhesive assay was performed by replanting EPCs on humanfibronectin-coated plates and then counting the adherent cells.The relationship of the EPCs' number and activity with Framingham Risk Score of ten years was also be assessed.Results Number of EPCs and the migratory & adhesive capacity were significantly lower in patients than in the control(P

Objective To construct a prokaryotic expression vector of cystatin C (Cys C), purify Cys C protein produced by the expression system, and prepare its antiserum. Methods Total RNA was isolated from HL-60 cells, and human Cys C gene was amplified with RT-PCR. The cDNA fragment was cloned into pMD18-T vector and which was confirmed by sequencing. The enzyme-digested target fragment was cloned into PET-32(a) expression vector and transfected into E.coli. BL 21(DE3), in which Cys C expression was induced. After the inclusion body protein was purified through Ni2+ affinity chromatography, processed by dialysis, identified by Western blotting, a rabbit was immunized with the fusion protein, and the antiserum was obtained. Results The result of DNA sequence analysis showed that the cloned Cys C gene sequence was completely corresponding to GenBank data. SDS-PAGE and Western blotting showed that the expressed Cys C fusion protein was about 35?103, mainly existing in the inclusion body of E.coli., that could be purified through Ni2+ affinity chromatography. The titer of the antiserum to the purified protein was 1∶8 000 by ELISA, and Western blotting confirmed that the antiserum reacted specifically to the Cys C protein. Conclusion A recombinant Cys C protein and the specific polyclonal antibody have been obtained, which provides a basis for establishment of immunoassays of human Cys C.

Objective To analyze the clinical and pathological characteristics of children with lupus nephritis(LN).Methods The data of 116 inpatients from Mar.2000 to Nov.2008 with LN were retrospectively analyzed.The clinical,immunochemical and pathological data were recorded.Renal tissue was observed by light microscopy and electron microscopy after HE,PAS,Masson and PASM staining according to WHO standards.Follow-up results showed complete remission,partial remission,disease activity,renal insufficiency and death.Results Of the 116 cases of LN,there were 27 male and 89 female with a ratio of male to female 1.03.3,and the mean age was(12.0?2.2) years.The incidence of nephrotic syndrome was 63.8 %(74 cases),and acute nephritis was 21.5%(25 cases).Class Ⅳ LN was the most frequent type(14 cases,50%) and classⅢ was next(25 cases,21.5%).In view of the outcome,rapidly progressive glomerulonephritis and class Ⅳ LN were the worst.LN was initially controlled in 96.5% of the patients.Relapses of LN were most caused by the intermittent treatment.Totally 32 cases showed different renal injury,and 6 cases progressed to death.Conclusions Renal biopsy is important to diagnosis,treatment and prognosis evaluation of LN.Long and regular treatment is important for children with LN.

0.05), but the lesion detection sensitivity of SSD and Raysum display were lower than that of UGI(? 2=4.17,7.11, and 5.14,4.17, P0.05). Excess fluid remained in the stomach and patient respiratory movement during breath holds were the reasons causing severe artifacts (6.1%) that influenced the diagnostic evaluation. Conclusion The performance of CTVG was equivalent to UGI in the detection of advanced gastric carcinoma and superior to UGI in the Borrmann′s classification. CTVG has potential in the detection of early gastric carcinoma. The value of SSD and Raysum display was limited in the evaluation of gastric carcinoma when used alone clinically.

Objective To reassess the value of CT and clinical criteria as prognostic and severity indicators in acute pancreatitis and to investigate the correlation between them.Methods Of 65 cases of acute pancreatitis,the hospitalization days,fevering days and entire complications (including local and systemic complications)were regarded as clinical endpoints.CT criteria included Balthazar's plain CT scan score,necrosis score,CT severity index(CTSI) and London's PSI score.Clinical criteria included Ranson and APACHE Ⅱscore.Using analysis of variance,t-test and multiple linear regression analysis,the correlation between each criteria and the three clinical endpoints was examined as well as the relation between CT and clinical criteria.The power of each criteria and combination of CT and clinical criteria in predicting entire complications of AP was assessed and compared by using ROC analysis.Results The mean scores of PSI,Ranson and APACHE Ⅱamong three groups classified according to entire complications were significantly different.Except Balthazar's plain CT scan criteria,each criteria's mean scores in group with local complications was signifiantly higher than that in group without and entire complications was significantly more in sever group than that in mild group.Mean hospitalization days and fevering days were significantly longer in sever group than that in mild group with Ranson Score.PSI and Ranson score had linear correlation with fevering days,and Ranson score had linear correlation with hospitalization days.In CT criteria,a linear correlation was only found between PSI and Ranson score.ROC analysis showed the Az of Ranson score was the largest,and there was no increase in the Az when CT criteria were added to clinical criteria.Conclusion The predictive value of Ranson score and PSI are superior to that of others.Clinical criteria is superior in predicting systemic complications,whereas CT is superior in predicting local complications.There is no improvement in predicting entire complications of AP when CT criteria are added to clinical criteria.The findings of plain CT scan is found to be some laggard compared with that of clinic.CT scan and short follow-up are important in the evaluation of AP.

OBJECTIVE:To establish a method for simultaneous determination of 6 residual organic solvents in aprepitant raw material as methanol,ethanol,acetone,isopropyl alcohol,methyl tert-butyl ether and tetrahydrofuran.METHODS:Headspace capillary gas chromatography was adopted.The determination was performed on DB-624 capillary column using temperature programming.The temperature of injector port was 180 ℃,and flame ionization detector was used with temperature of 260 ℃.Nitrogen was used as carrier gas with flow rate 3.0 mL/min.The spilt ratio was 5 ∶ 1,and head-space injection volume was 1.0 mL.The head-space equilibrium temperature was set at 80 ℃,and equilibrium time was 40 min.RESULTS:The linear ranges of methanol,ethanol,acetone,isopropyl alcohol,methyl tert-butyl ether,tetrahydrofuran were 6.052-605.232 μ g/mL (r=0.999 9),9.987-998.718 μg/mL(r=0.999 9),9.998-999.768 μg/mL(r=0.999 8),9.986-998.634 μg/mL(r=0.999 9),9.991-999.090 μg/mL (r=0.999 7),1.461-146.133 μg/mL(r=0.999 5),respectively.The limits of quantitation were 1.782 1,2.079 0,0.749 8,1.777 8,0.223 1,0.607 0 μg/mL;the limits of detection were 0.594 0,0.693 0,0.249 9,0.592 6,0.074 4,0.202 3 μg/mL,respectively.RSD of precision test was lower than 2.0％.Only acetone and isopropyl alcohol were detected in stability test and reproducibility tests,RSD＜2.0％.Their recoveries were 99.34-100.75％ (RSD=0.52％,n=9),98.20％-100.24％ (RSD=0.69％,n=9),98.07％-100.07％ (RSD=0.84％,n=9),99.86％-101.32％ (RSD=0.58％,n=9),97.87％-104.02％ (RSD=2.13％,n=9),98.26 ％-100.58 ％ (RSD =0.75 ％,n =9),respectively.CONCLUSIONS:The established method is simple,accurate and reproducible,and can be used for simultaneous determination of 6 residual organic solvents in aprepitant raw material.

Objectives To investigate the effects of lipoxin receptor agonist BML-111 on IFN-βand IE86 mRNA expression of macrophages infected by human cytomegalovirus (HCMV). Methods Macrophages were infected with HCMV (MOI=0.5), and the cultured cells were randomly divided into control group, HCMV group, HCMV+BML-111 group, and HCMV+MP group. The cells were collected at 0,1,2,4,8 and 12 h after infection, and the levels of IFN-βand IE86 mRNA were tested by real-time PCR. Results Compared with HCMV group, the levels of IFN-βmRNA in HCMV+BML-111 group increased significantly (P < 0.05), while the levels of IFN-βmRNA in HCMV+MP group decreased significantly (P < 0.05); Compared with HCMV group, there were no significant differences of the levels of IE86 mRNA in HCMV+BML-111 group (P>0.05), while the levels of IE86 mRNA in HCMV+MP group increased significantly (P < 0.05). Conclusion BML-111 exerts antiviral activity by promoting the expression of IFN-βmRNA at the early stage of HCMV infection.

BACKGROUND:The anti-inflammation and protective effects of lipoxin have been verified in several immunity-related disease models. Preliminary studies of our research group have shown that, lipoxin receptor agonist BML-111 has negative regulation effects on the human cytomegalovirus (HCMV)-induced immunological injury. However, the effect of BML-111 on the HCMV replication remains unclear.
OBJECTIVE:To observe the influence of lipoxin receptor agonist BML-111 on HCMV replication and proliferation in THP-1 macrophages and human embryonic lung fibroblasts.
METHODS:THP-1 macrophages were infected by HCMV AD169 strain, and were divided into three groups:mock infection, HCMV infection, HCMV+BML-111. The final concentration of BML-111 was 100 nmol/L. cells in each group were col ected at 0, 1, 2, 4, 12, 36, 48 hours, the mRNA levels of IE86 and pp65 in the THP-1 macrophages were tested by RT-PCR method. Human embryonic lung fibroblasts were infected with HCMV (MOI=0.1), and were divided into two groups:HCMV infection and HCMV+BML-111. The patho-morphous changes of human embryonic lung fibroblasts were observed under light microscope, and the cellnumber was measured. The infective virus titer changes in human embryonic lung fibroblasts were examined by plaque assay.
RESULTS AND CONCLUSION:After the macrophages were infected by HCMV, compared with the mock infection group, the mRNA levels of IE86 and pp65 in the HCMV group and HCMV+BML-111 group were increased significantly;compared with the HCMV infection group, the mRNA levels of IE86 and pp65 in the HCMV+BML-111 group were increased significantly in the early stage (within 4 hours) after infection, but the pp65 mRNA levels were decreased significantly in the medium and late stages (24-72 hours) after infection. After human embryonic lung fibroblasts were infected by HCMV, the degree of the patho-morphous in the HCMV+BML-111 group reached 100%2 days earlier than the of HCMV infection group. The infective virus titer reached the peak 2 days earlier than the HCMV infection group, but no significant difference was found between the two groups. BML-111 accelerates the replication of HCMV in the early stage of infection, but inhibits the expression of pp65 gene in the late stage. BML-111 has no impact on the proliferation of the infective HCMV titer in vitro.

Objective To investigate the effects of exogenous wild DCC gene stably transfection on growth of colorectal carcinoma cell line SW1116 in vitro. Methods DCC gene domain was amplified from human normal colon tissue by reverse transcript-polymerase chain reaction(RT-PCR). At first,a recombinant expression plasmid pcDNA3. 1( + )-DCC was constructed. Human colorectal carcinoma cell line SW1116 with-out DCC gene was transfected with pcDNA3. 1-DCC. Cell viability was tested by methyl thiazolyl tetrazolium (MTT)assay. Immunofluorescence staining was used to determine the effects of pcDNA3. 1-DCC and expres-sion of carcino-embryonic antigen(CEA)in human colorectal carcinoma cell line SW1116 which was transfect-ed with pcDNA3. 1-DCC. Results The population of cells transfected with pcDNA3. 1( + )-DCC plasmid was lower than those with pcDNA3. 1( + )-DCC plasmid and normal cells(t = 3. 645,P ﹤ 0. 05;t = 3. 132,P ﹤0. 05)at 3 ~ 6 days after transfection,and the proliferation rate of cells transfected with pcDNA3. 1( + )-DCC plasmid was lower than those with pcDNA3. 1( + )plasmid and normal cells(t = 2. 134,P ﹤ 0. 05;t = 2. 736, P ﹤ 0. 05). Cell line SW1116 transfected with pcDNA3. 1( + )-DCC plasmid total viability was lower than nor-mal cells(t = 3. 053 ,P ﹤ 0. 05)at 2 ~ 6 days after transfection. Cell line SW1116 transfected with pcDNA3. 1 ( + )-DCC plasmid total viability was lower than those with pcDNA3. 1( + )plasmid(t = 2. 816,P ﹤ 0. 05)at 2,4,5,6 days after transfection. The population of flavo-green colour cells transfected with pcDNA3. 1( + )-DCC plasmid and the fluorescent intensity of these cells were lower than those with pcDNA3. 1( + )plasmid and normal control cells. Conclusion Transfected DCC gene can suppress the cell proliferation and make CEA expression of cell line SW1116 down regulation to weaken its infiltration and metastasis abilities.

Purpose Lipotoxicity plays an important role in the progression of diabetes and its complications,and the gold standards for organs' lipid quantification are biopsy or histochemical staining,which are invasive and have their own limitations.Thus,our research was to identify the difference in organ lipid deposition between type 2 diabetic patients and healthy volunteers by using noninvasive three-point T2* corrected Dixon imaging.Materials and Methods Sixty-five type 2 diabetic patients and 34 healthy volunteers were included in this study.All participants underwent three-echo Dixon with 3.0T MR imager.Dixon imaging parameters [hepatic fat fraction (HFF),splenic fat fraction (SFF),pancreatic fat fraction (PFF)] were collected.All of the MRI parameters were compared.The relationship between HFF,SFF,PFF and BMI,age were analyzed.Results The hepatic and splenic lipid percentage in diabetic group [HFF=(5.4±4.3)％;SFF=(3.7± 1.4)％] was significantly higher than healthy volunteers group [HFF=(2.9± 1.3)％;SFF=(3.0± 0.9)％;P＜0.05,respectively].However,there was no significant difference in pancreatic fat fraction between diabetic and healthy groups (P＞0.05).There were positive correlations between HFF,SFF,PFF and BMI (r=0.379,0.305 and 0.306,P＜0.05).Moreover,only the positive correlation between pancreatic fat fraction and age were observed (r=0.261,P＜0.05).Conclusion The three-point T2* corrected Dixon revealed the abnormalities of hepar and spleen lipid accumulation in diabetic patients.Thus,the three-point Dixon imaging may potentially aid in evaluating the lipid deposition of abdominal organs.