Objective To investigate the changes of monocyte chemoattractant protein-1(MCP-1)in serum and urine of lupus nephritis(LN)patients in active phase and remission phase.Methods The levels of MCP-1 in serum and urine of 58 LN patients(27 of active phase and 31 of remission phase)were measured by ELISA.The correlation between the levels of MCP-1 in variant phase of LN and other relevant factors were analyzed.Results The MCP-1 levels in sera of both active phase and remission phase of LN patients were markedly higher than those in controls(548.5?347.2 ng/L and 469.1?298.4 ng/L vs 273.3?146.7 ng/L,P0.05).Conclusion The MCP-1 levels in urine of LN patients is more suitable to evaluate the activity of disease as a sensitive marker.

Objective To quantify the level of human sperm protein 17(Sp17)in serum of the healthy and patients with gynecological cancers and to evaluate whether Sp17 is a suitable serum marker for the diagnosis of gynecological cancers.Methods The recombinant human Sp17 was radioiodinated by the chloramine T method.125I-rhSp17 was used to develop a radioimmunoassay to determine Sp17 in serum of the healthy and the patients.Meanwhile,Sp17 levels in serum were compared with the expression of Sp17 in tissues from gynecological cancers.Results The level of serum Sp17 was(15.60?7.66)?g/L in healthy subjects(n=59),which was significantly lower than those in twenty-two patients(64.36?34.44)?g/L.Sera from nine of twenty-two patients(40.9%)were positive for Sp17 by radioimmunoassay assay.Tumor tissues from twelve of twenty-two patients(54.5%)were positive for Sp17 by immunohistochemistry assay.Conclusions Markedly elevated serum levels of Sp17 were found in patients with gynecological cancers.However,there was no significant correlation between serum level of Sp17 and expression of Sp17 in tumor tissues in patients with gynecological cancer.

Objective To study the effects and possible mechanism of Vitamin K2(Vit K_2)in the treatment of K562 cells.Method The possible mechanism of the apoptosis of K562 cells induced by Vitamin K2(Vit K_2)were investigated with the transmission electron microscope,flow cytometry(FCM),retrotrans criptase polymerase chain reaction(RT-PCR)and chemiluminescence assay.Results Apoptosis peak on FCM and positive AnnexinV-FITC on cell membrane showed that Vit K_2 induced apoptosis of K562 cells and in a dose-and-time-dependent manner;G0/G1 cell arrested;significantly down-regulated the expression of bcl-2 and survivin,but had no effect on the expression of bax;The activities of caspase-3 were significantly increased.Conclusions Vit K_2 induces apoptosis of K562 cells through activating caspase-3 pathways and the apoptosis-related genes bcl-2、survivin might play an important role in this process.

Objective To observe the changes of HNP1-3 and NE content in serum quantitatively during perioperation of cardiac surgery under cardiopulmonary bypass(CPB),and to explore the role of changes of HNP1-3 and NE content in early diagnosis of system inflammatory response syndromes(SIRS)after cardiopulmonary bypass.Methods The contents of HNP1-3 and NE in serum during perioperation of 21 cardiac surgeries under cardiopulmonary bypass were measured quantitatively using ELISA method,and the contents of CK,CK-MB and C-RP in serum were measured at the same time points as that of HNP1-3 and NE.Results The contents of HNP1-3 and NE in serum were 54.55?26.45 ng/ml and 41.09?9.93 ng/ml respectively before CPB,and they were 435.45?500.13 ng/ml and 250.91?97.21 ng/ml respectively 0.5 h after CPB which were significantly higher than that before CPB(P

Objectives To establish a reliable method for isolating and purifying soluble collagen type Ⅱ(SCⅡ)by optimizing preparative procedure.Method The chicken sternal cartilage was selected as raw material.Guanidine hydrochloride was used to remove the proteoglycans.The digestion manners of pepsin,sodium chloride concentrations for salting,types of DEAE anion resin were studied for extracting SCⅡ.The SCⅡ identification was made by SDS-PAGE,absorption spectrum and amino acid analysis.Result It was convenient for pre-treatments of chicken sternal cartilage.The proteoglycans could be efficiently removed by 4 mol/L guanidine hydrochloride.The satisfied results were obtained by limited enzyme digestion of pepsin added by two steps.The optimizing concentration of sodium chloride for salting was 2.4mol/L.SDS-PAGE maps revealed that the bands of purified SCⅡand standard CⅡ were at the same location.The absorption peak of SCⅡ was at 230nm.The concentrations of Gly,Pro and Ala were the highest in 15 amino acids.Conclusion The improved method has significant advantages of simple working process,result reliability and convenient source of raw material.It is suitable for purifying the SCⅡ at variable scales in research works and clinic application.The SCⅡ product obtained has high purity and accords with the characteristics of collagen type Ⅱ.

Objective To optimize the conditions of the expression of fusion protein GST-SjMP10 and to evaluate the value of fusion protein GST-SjMP10 for diagnosis of schistosomiasis.Methods The optimal concentration of IPTG for the expression of fusion protein GST-SjMP10 was chosen in inducing the expression of GST-SjMP10 with different concentration of IPTG,and the soluble fusion protein GST-SjMP10 was identified by SDS-PAGE.The fusion protein GST-SjMP10 was purified by chromatographic affinity with glutathione Sepharose 4B gel.The sensitivity and specificity of purified fusion protein GST-SjMP10 for diagnosis of schistosomiasis were determined by enzyme linked immunosorbent assay(ELISA)to detect the IgG antibody in sera from the patients with acute schistosomiasis,advanced schistosomiasis and clonorchiasis as well as healthy subjects.Results Most of the expressed fusion protein GST-SjMP10 was in soluble status when the concentration of IPTG was reduced to 0.1 mmol/L and the fusion protein GST-SjMP10 could be purified by chromatographic affinity.The positive rate of anti-GST-SjMP10 antibody in the sera from the patients with acute and chronic schistosomiasis japonica was 97.5% and 96.7% respectively.No cross reactivity of the fusion protein GST-SjMP10 was found in the detection for the sera from clonorchiasis patients,and no false positive was found in the detection for the sera of healthy subjects.Conclusion The fusion protein GST-SjMP10 was expressed successfully and showed high sensitivity and specificity for the diagnosis of schistosomiasis japonicum.

Objective To develope a new kind of piezoelectric immunosensor which could detect anti-HCV antibody in short time.Methods The corresponding sensitive material was fixed on the piezoelectric immunosensor by polyethylenimine adhesion and glutaraldehyde cross-linking method.The assembly condition and response characters were investigated and the piezoimmunosensor could be used in detection of clinical blood specimen.Results The respond of the positive serum was 4 times higher than that of negative serum.HBsAg had little interference to the detection of anti-HCV antibody.Conclusion The piezoelectric immunosensor prepared in this study is highly selective,easy to be operated for effective detection of anti-HCV antibody.

Objective To develop a PCR-SSP method for detection of HLA-DRB1 alleles in the patients who were hypersensitive to Platanus Acerifolia pollen allergen,and to probe into the association between the atopic subjects to Platanus Acerifolia pollen allergen and HLA-DRB1 alleles.Methods DNA in whole blood was extracted by phenol-chloroform method.Eight pairs of specific primers for alleles were synthesized,and HLA-DRB1*0401,*0402,*0403,*0404,*0405,*0406,*0407,*0408 alleles in 20 atopic patients and 36 healthy individuals of Jiangsu Province with Han nationality were detected by PCR-SSP(polymerase chain reaction-sequence specific primer).Results By optimizing the experimental conditions PCR-SSP methods for detection of the 8 alleles were established and the distributing data of above-mentioned HLA DRB1 were obtained.The frequency of HLA DRB1*0405 and *0406 in the patients group was higher than that of in healthy controls group,while the frequency of HLA DRB1*0402 in the patients group was lower than that in controls.No significant deference for the other 5 alleles was found between the 2 groups.Conclusion HLA-DRB1*0406和*0405 seems to be the likely suspected candidate alleles responsible for susceptibility to Platanus Acerifolia pollen allergen in the atopic patients,while DRB1*0402 might be contribute to the related resistance to the allergen.

Objective To determine the biological reference interval of urinary conductivity in healthy people and to study the relationship between urine conductivity and other parameters as well as its clinical feasibility.Methods Conductivities of midstream urine specimen from healthy people (n=10119,5074 males and 5045 females,aged under 96 years) or patients with different diseases (n=3449) were determined.Among them the following parameters:conductivity,osmolality,specific gravity,creatinine,urea,uric acid,sodium,potassium and chloride of 250 random urine specimens were simultaneously determined.Results The urinary conductivities in healthy people exhibited normal distribution and significant differences were found in the subjects with different age and sex.The reference range of urinary conductivity was between 10.42?4.61 mS/ch and 24.10?6.81 mS/ch in healthy people and between 7.95 mS/ch?2.40 and 18.01?5.90 mS/ch in the patients with different diseases.Conductivity determined was positively correlated with osmolality (r:0.894),specific gravity (r:0.727),sodium (r:0.698),potassium (r:0.563),chloride (r:758),uric acid (r:0.521),urea (r:0.556) and creatinine (r:0.495).Conclusions Urine conductivity,which determination is simple and rapid,may reflect the conductive capacity of electrolytes in urine and positively correlated with osmolality,so it can be used as a new parameter for urinalysis to diagnose renal concentrative function in routine laboratory.

Objective To evaluate the correlation of intrahepatic HBV DNA load with the HBV load in serum and peripheral blood mononuclear cells(PBMC)and the stage of fibrosis,grade of inflammation,level of serum ALT and AST.Methods Liver specimens were taken from 50 patients by percutaneous needle biopsy and divided into two parts:one was processed for histological examination,and the other was used for molecular biology analyses.HBV DNA load was measured with fluorescent quantitative polymerase chain reaction(FQ-PCR).The data of serum level of ALT and AST were collected.Results A high correlation between intrahepatic HBV-DNA load and serum virus load was found(r=0.77977,P