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Serum amyloid A(SAA) is a novel marker widely used in the acute infection disease, especially viral infection. SAA has shown a cerntain value in assisting the clinical diagnosis, discrimination of severity and monitoring of progress and outcome of COVID-19. This paper introduces the application of SAA structural, function andits dynamic detection in the diagnosis of COVID-19, and the significance of combined detection with COVID-19 antibodies, nucleic acid and other diagnostic indicators.
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Objective To study the characteristics of bone marrow hematopoietic recovery such as absolute neutrophil count (ANC) and platelet (PLT ) after accepting hematopoietic stem cell transplantation in the patients with inv (9) .Methods A total of 39589 cases of definitely diagnosed hematonosis in our hospital from January 2010 to October 2015 served as the research subjects . The R banding technique ,polymerase chain reaction(PCR) and flow cytometry instrument were adopted to check chromosome karyotype ,fusion gene and bone marrow hematopoietic recovery related indicators .Results PML-RARα,BCR-ABL1 ,AML-ETO , EVI1 ,CBFβ-MYH11 ,MLL-AF6 ,AML-AF4 ,SET-NUM214 ,SIL-TALI ,IgH rearrangement ,TCR rearrangement and BCL1-IgH and other fusion gene were detected in the patients with inv (9) hematonosis .The recovery situation after receiving hematopoietic stem cell transplantation in the patients with inv (9):ANC recovered to >0 .5 × 109/L on 12 d after transplantation ,PLT recovered to >20 × 109/L on 16 d after transplantation .The recovery situation after receiving hematopoietic stem cell transplantation in the patients with noninv(9):ANC recovered to >0 .5 × 109/L on 12 d after transplantation ,and PLT recovered to >20 × 109/L on 13 d after transplantation .Conclusion The time achieving ANC recovery >0 .5 × 109/L after hematopoietic stem cell transplantation in the patients with inv(9) and without inv(9) is almost similar ,while the time achieving PLT count recovery in the patients with inv(9) is slightly longer than that in the patients without inv(9) .
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Objective To research the mechanism of the changes of T lymphocyte subtypes and provide reference for clinically prevention,diagnosis and treatment for NSCLC through analysis of the expression of Th1 ,Th2 in Non-small-cell carcinoma (NSCLC)patients.Methods Whole blood (EDTA anticoagulant treatment)from 60 NSCLC patients and 60 healthy sub-j ects were collected to detect of the expression of CD3 +T cells,CD4+T cells and CD8+T cells on T lymphocytes and the lev-els of Th1 and Th2 cells by flow cytometer (FCM),and the absolute value of T lymphocyte by hematology analyzer.Results Compared with normal control group,after surgery 1~3 days NSCLC groups,the percent of CD3 +T,CD4+T,CD8+T cells and the CD4+/CD8+ ratio in the NSCLC patients before surgery were significantly reduced 58.40±10.27 vs 66.58± 6.84,31.32±8.65 vs 39.40±6.43,34.23±8.00 vs 24.31±8.16,0.96±0.23 vs 1.58±0.23 (t=-6.726~14.916,P<0.05).The percent of CD3 +T,CD4+T,CD8+T cells and the CD4+/CD8+ ratio in the NSCLC patients after surgery 1~3 days were also significantly decreased 56.31±8.00 vs 66.58±6.84,27.72±7.55 vs 39.40±6.43,33.69±7.10 vs 24.31± 8.16,0.87±0.31 vs 1.58±0.23 (t=-6.720~14.367,P<0.05).The percent of CD4+T cells in the NSCLC patients af-ter surgery 4~7 days was increased 33.23±4.13 vs 39.40±6.43(t=6.257,P<0.05).Compared with the control group, within the helper T cell subsets,the cell content of Th1,Th2 cells (× 10/μl)and the Th1/Th2 ratio were significantly changed in different extent in the NSCLC group before surgery 6.79±1.34 vs 12.52±3.56,4.82±0.51 vs 2.32±0.82, 1.39±0.84 vs 5.36±1.42 (t=-20.087~18.630,P<0.05).The content of Th1 cells was lower in the NSCLC patients after 1~3 days and 4~7 days 8.86±1.52 vs 12.52±3.56,7.02±1.27 vs 12.52±3.56 (t=7.339~11.275,P<0.05). Conclusion The NSCLC patients presented immune dysfunction,like T lymphocytes and helper T cells decreased and Th2 cells were clearly in the ascendant.Also,the cytotoxic T cells increased by the stimulation of cancer cells,but they began to decrease after the surgery.
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Objective To analyze and evaluate the expression and significance of costimulatory molecules CD 28 ,CTLA-4 ,CD86 , CD80 mRNA in peripheral blood mononuclear cells of the patients with HBV chronic change .Methods The levels of costimulatory molecules CD28 ,CTLA-4 ,CD86 ,CD80 mRNA in peripheral blood mononuclear cells were detected in 24 cases of chronic hepatitis B (CHB) ,24 cases of liver cirrhosis(LC) ,28 cases of hepatocellular cancer(HCC) and 30 normal control(NC) subjects by real time quantitative PCR .Results Compared with the NC group ,costimulatory molecule CD28 mRNA level in the CHB group was signifi-cantly decreased(t= -2 .11 ,P<0 .05);CTLA-4 mRNA level in different diseases groups was decreased to different degrees :the CHB group(t= -2 .52 ,P<0 .05) ,the LC group(t= -2 .11 ,P<0 .05) and the HCC group(t= -2 .56 ,P<0 .05);CD86 mRNA level in different diseases groups was decreased to different degrees too :the CHB group(t= -3 .68 ,P<0 .01) ,the LC group(t= -2 .99 ,P<0 .01) and the HCC group(t= -4 .42 ,P< 0 .01);CD28/CTLA-4 mRNA level was significantly increased in the HCC group(t= 2 .12 ,P< 0 .05);CD80/CD86 mRNA level was significantly increased to different degrees with the progress of HBV chronic change:the CHB group(t=2 .10 ,P<0 .05) ,the LC group(t=2 .59 ,P<0 .05) and the HCC group(t=3 .74 ,P<0 .01) . Conclusion The expression abnormality of CD28/B7 family costimulatory molecules mRNA in HBV infectious patients may be closely related with the immune dysfunction and the development and progression of the chronic change .
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Objective To investigate the expression and significance of CD28,cytotoxic T-lymphocyte antigen-4 (CTLA-4),programmed death-1 (PD-1) and T cell immunoglobulin mucin-3 (Tim3) on T lymphocytes in chronic HBV-infected patients.Methods A total of 102 chronic HBV-infected patients,including 42 patients with chronic hepatitis B (CHB),30 patients with hepatitis B-induced liver cirrhosis (LC),and 30 patients with hepatocellular carcinoma (HCC),were enrolled from the First Affiliated Hospital to Soochow University during October 2012 and June 2013.Thirty healthy individuals were also enrolled as controls.Expression of CD28,CTLA-4,PD-1,Tim-3 on T lymphocytes in peripheral blood were determined by flow cytometry,and the differences among groups were analyzed using one-way ANOVA and LSD-t test.Spearman correlation test was performed to analyze the correlations of the expression of CD28,CTLA-4,PD-1,Tim-3 on T lymphocytes with HBV DNA loads,HBeAg and ALT.Results The expression of CD4 + CD28 +,CD8 + CD28 +,CD4 + CTLA-4 + in chronic HBV-infected patients were lower than those in healthy controls.CD4 + CD28 + expression in HCC group was lower than that in CHB group (t =2.373,P < 0.05) ; CD8 + CD28 + expression in LC and HCC group was lower than that in CHB group (t =4.324 and 4.088,P < 0.01) ; CD8 + PD-1 +,CD4 + Tim-3 + and CD8 + Tim-3 + expressions in CHB group were higher than those in LC,HCC group and healthy controls (t =3.051,3.130,3.121,3.254 and 3.723,P <0.01).CD8 + PD-1 + expression was positively correlated with ALT levels and HBV DNA loads (r =0.516 and 0.582,P < 0.01) ; CD8 + Tim-3 + expression was also positively correlated with ALT levels andHBV DNA loads (r =0.578 and 0.556,P <0.01); PD-1 and Tim-3 expressions on CD8 T lymphocytes were positively correlated with each other (r =0.578,P < 0.01).Conclusion The abnormal expression of the molecules on T lymphocytes in chronic HBV-infected patients is closely correlated with immune function disorder and the progression of the disease.
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Objective To investigate the significance of changes in expression of co-stimulatory molecules on T lymphocytes in patients with chronic hepatitis B (CHB) infection.Methods In a casecontrol study,a total of 82 CHB cases including 50 male cases and 32 female cases (the mean age was 42.32 ±3.74) were enrolled in the First Affiliated Hospital to Soochow University from October 2012 to November 2013,together with 30 cases health control (15 male cases and 15 female cases,and the mean age was 42.32 ± 3.74).Patients were divided into three groups:40 cases of non-treated,30 cases of effective-treated and 12 cases of ineffective-treated with anti-viral drugs and immune-related therapy.The expression levels of CD28,CTLA-4,PD-1,Tim-3 on T cells subset from peripheral blood were determined by Flow Cytometry.Serum load of HBV DNA was detected by Real-time PCR and the serology markers such as HBeAg and ALT were detected by conventional methods The Kruskal-Wallis test was used to analysis groups comparison.Independent samples t-test and Mann-Whitney U test were used to two sample comparison.Spearman's rank correlation test was used to analyze the correlation.Results The expression levels of CD28,CTLA-4 and PD-1 on CD4 in health control group was the highest compared to ineffectivetreated group [404.65 (331.65-536.09) vs 277.15 (249.90-344.25) (H=29.81,P<0.001);32.89 (29.69-39.69) vs 19.26 (11.90-20.56) (H =43.13,P <0.001),respectively].The expression level of PD-1 on CD8 in pre-treated group was the highest,while expression level in health control group was the lowest [15.47 (12.50-17.78) vs 3.05 (1.41-3.97) (H=56.60,P<0.001)].Similarly,the expression level of Tim-3 on CD4 in ineffective-treated group was the highest,while Tim expression on CD4 in health control group was the lowest [199.62 (55.61-239.45) vs 70.62 (53.88-112.32) (H =41.03,P < 0.001)].The expression level of Tim-3 on CD8 in pre-treated group was the highest,while it was the lowest in health control group [82.50 (78.69-84.58) vs 3.07 (1.56-6.87) (H=74.84,P <0.001)].Compared to the group with low viral load,the expression level of PD-1 on CD8 both in the pre-treated group with high viral load and the post-treated group was significantly increased [17.87 (13.38-20.94) (U=25.00,P<0.001); 16.95 (5.39-18.27) (U=63.50,P<0.001),respectively].Importantly,in the post-treated group,the PD-1 expression on CD4 T (r =0.689,P <0.001) and on CD8 T (r =0.751,P < 0.001) was also positively correlated with ALT.Conclusions The abnormal expression of these co-stimulatory molecules may be present in the antiviral treatment process of CHB.It may provide new clues for the reasonable treatment of CHB.
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Objective To investigate the change of expression patterns of miRNA in the serum and peripheral blood mononuclear cells (PBMC) of lung cancer and its significance.Methods Clinical case control study was employed.Establish the method of microRNA(miRNA) detection by real time quantitative PCR (RT-qPCR).peripheral blood of the study subjects were collected in First affiliated hospital of SooChow University from November 2011 to September 2012.Gender and age matched subjects whose median age was 64(40-85) included 61 lung cancer cases,48 healthy control and benign lung diseases.We used quantitative RT-PCR to assess miRNA expression pattern of-miR-20a,21,-25,-29,-31,-126,-129,-145 and -205 in peripheral blood.U6 was taken as reference,and the expression of miRNA were indicated as F =2-△Ct,ACt =CtmiRNA--CtU6.F represents relative change of miRNA expression compared to U6 in the same sample.SPSS 19.0 was used as statistical software; t test was used for comparison of two sets of samples One way ANOVA was used for multiple groups' comparison,and make multiple comparison by the S-N-K method if the result with a significant difference.Pearson correlation analysis were used for the relationship between two variables,Brown-Forsythe test was used for Ct value equality testing among multiple samples.P < 0.05were regarded as statistically significant.Results miR-20a (F =271.64,P < 0.01),miR-21 (F =2232.51,P<0.01),miR-205 (F=45.13,P<0.01),miR-29a (F=19.98,P <0.01),miR-25 (F=313.19,P < 0.01) and miR-126 (F =32.38,P < 0.01) were differently expressed in the serum of lung cancer patients and healthy control or benign disease control.miR-29a,miR-25,miR-126 was down regulated in the development of malignant lung disease; miR-31 elevated in lung cancer compared with healthy control,while miR-145 fell; miR-31 expression changed with various differentiation of lung cancer (F =5.22,P < 0.01)itwas significantly increased in the moderate-differentiated cancer,but decreased when distant metastasis existed (especially bone metastasis).But in PBMC paired with the serum samples above,statistically significance was shown in lung cancer and healthy control group and benign lung diseases group in miR-126 (F=690.58,P<0.01),miR-129 (F=26.66,P<0.01),miR-145 (F=48.57,P<0.01),miR-205 (F=308.61,P<0.01).miR-25 (F=218.57,P<0.01) and miR-31 (F=48.05,P<0.01),were down regulated in the development of malignant lung disease.miR-20a,miR-29a were elevated in lung cancer compared to healthy controls,miR-21 was up-regulated when distant metastasis existed; the expression of miR-31 in serum and PBMC was negatively correlated (r =-0.369,P < 0.05).Areas under ROC curve of miR-25 (S =0.906,P < 0.01) and miR-126 (S =0.969,P < 0.01) were statistically different.Conclusions miRNA may contribute to several steps of metastasis,including local invasion,extravasation or initial survival at a distant site,and metastatic colonization,or can affect the prognosis of lung cancer.The detection of miRNA in lung cancer provides a new clue to the research of its chronic progress.
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Clinical laboratory information system is the key to the realization of total laboratory automation,standardization,intelligent and digitization.In recent years,with the establishment,application,popularization and upgrading of clinical laboratory information system,it is very important to establish the construction criterion of clinical laboratory as soon as possible.Now according to the requirements of construction and management of modern clinical laboratory and combining with the practical application of clinical laboratory information system in our hospital at domestic settings,the standardization construction of clinical laboratory information system was discussed in this article.
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Objective To explore the significance and the change of miRNAs expression profile in different period of HBV infection.Methods Establish the detection method of microRNA(miRNA) by RNA DNA probe liquid chip (flexible multi-analyte profiling,xMap) and estimate the specificity,repeatability and accuracy of this method.From October 2010 to October 2011,collect HBV infected patients' periphcral blood of the First affiliated hospital of SooChow University,including acute hepatitis B,chronic hepatitis B,hepatits B liver cirrhosis,liver cancer patients and health control,each group contains 40 cases.The levels of miR-191,-223,-222,-145,-21,-31,-126,-20a,-372 of peripheral blood mononuclear cell were detected by xMap liquid chiptechnology.The level of miR-103 was taken as reference.The ratio of (the mean fluorescence intensity of target miRNAs-corresponding backgroud mean fluorescence intensity)/( the mean fluorescence intensity of miR-103-corresponding backgroud mean fluorescence intensity) as a valid data and analysis the characteristics of miRNAs expression.The SPSS 17.0 was used as statistical software.The single factor analysis of variance was used as the method to analysis group comparison,and make multiple comparison by the LSD-t method if the result with a significant difference.Results The specificity of Xmap liquid chip method to detect miRNAs was 100% ;The repeated experiment proved that the CV value was less than 5% in the high value reference miRNA test,less than 10% in the low value reference miRNAs test;the accruracy experiment proved that the recovery rate was ( 100 ± 5)% in the nine miRNAs.There were no statistically differences with miR-222 ( F =1.32,P > 0.05),-191 ( F =1.98,P > 0.05),-145 ( F =0.78,P>0.05),-21(F=0.64,P >0.05),-31 (F =0.83,P >0.05),-372(F =1.75,P >0.05)in different groups; There was statistically significant differences in miR-223 ( F =14.56,P < 0.05) among different groups,with the highest expression level in actue hepatitis B group (15.37 ± 4.01),and the lowest expression level in liver cancer group (6.91 ±3.18) ; There was statistically significant differences in miR126 (F =17.43,P < 0.05)among different groups,with the highest expression level in health control group (6.33 ±2.75) and the lowest expression level in liver cancer group (2.38 ± 1.07).There were statistically significant differences in miR-20a ( F =19.484,P < 0.05) among different groups,with the highest expression level in health control group (0.33 ±0.18) and the lowest expression level in liver cancer group (0.81 ±0.24).Conclusion The detection method of miRNA by Xmap liquid chip has strong specificity,high accuracy and good repeatability,suitable for large throughput clinical testing.The study for miRNA in HBV infected diseases provides a new clue to the research of chronic progress mechanism.
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Objective To evaluate and report the performance of PL-11 platelet analyzer.Methods Intravenous blood samples anticoagulated with EDTA-K2 and sodium citrate were tested by the PL-11 platelet analyzer to evaluate the intra-assay and inter-assay coefficient of variation (CV),carry-over rate,accuracy,linearity of the PL-11 platelet analyzer.Platelet aggregation rate of sodium citrate-anticoagulated fasting venous blood collected from 30 physical examinees in outpatient department of The First Affiliated Hospital of Soochow University between February and July,2012 was detected by the PL-11 platelet analyzer and MPG-3E multifunctional double channel blood coagulation analyzer,respectively.The correlation was detected between the PL-11 platelet analyzer and MPG-3E multifunctional double channel blood coagulation analyzer.Results All the parameters were conformed to the standard of Clinical Laboratory Improvement Amendment 88.Both of the intra-assay and inter-assay CV values were less than 5% ; carry-over rate was less than 1% ; the accuracy and the linearity was excellent correlated to the result of photoelectric turbidimetry (R2 =0.9439).Conclusions PL-11 platelet analyzer can directly use whole blood dynamics to analyze platelet aggregation process and quantitatively analyze the various components of blood cells (including platelets and red blood cells),the result reports are accurate and reliable.
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ObjectiveTo explore the relationship between the negative co-inhibitor programmed death-1 ( PD-1 ) and the pathogenesis of myasthenia gravis ( MG), by detecting the expression of PD-1 and programmed death ligand-1 ( PD-L1 ) on peripheral blood mononuclear cells (PBMCs) and soluble PD-1 (sPD-1) in plasma from myasthenia gravis patients. MethodsPeripheral blood samples were collected from 45 MG patients and 33 healthy persons without prednisone or other immunodepressant treatment during the half year ahead of withdrawal.The expression of PD-1 and PD-L1 on PBMCs were detected using immuno-fluorescence labeling and flow cytometry, and the concentrations of sPD-1 in plasma were measured using an ELISA kit. Results(1) The proportion of CD4+ PD-1 + T cells, as well as CD14+ PD-L1 +monocytes of the MG group was higher than that of the control group. There were no significant differences in the proportion of CD4+ PD-1 + T cells or CD14+ PD-L1 + monocytes in the MG sub-groups between different genders or MG types. While the proportion of CD4+ PD-1 + T cells of the late-onset MG (age ≥40) group was higher than that of the early-onset MG group (age <40). And it was higher in the MG patients with thymoma or thymus hyperplasia than that from the MG patients with normal thymus. The proportion of CD14+ PD-L1 +monocytes from the MG patients with thymoma or thymus hyperplasia group decreased obviously compared with that of the patients with normal thymus group; but no difference could be found between the late-onset group and early-onset group. (2)The concentration of sPD-1 in the plasma from the group of MG patients was(6. 92 ±0. 72) ng/ml,which was higher than that of the healthy control group ( (3.28 ±0. 42) ng/ml),even more, it was significantly higher in the early-onset MG group than that of the late-onset MG group,there was a negative correlation( r =-0. 526, P =0. 000) between the age of onset and the concentration of sPD-1. ConclusionsThe increased expressions of PD-1 on CD4+ T cells and PD-L1 on CD14+ monocytes in MG patients suggested the involvement of the couple of molecules in the pathogenesis of MG.Higher concentration of soluble PD-1 in the plasma of patients with MG suggested that it might disturb the ligation of PD-1 and PD-L1 on T cells and antigen presenting cells, which might result in the abnormal transportation of the negative modulating signal, and accelerate the pathological progress of MG.
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Objective To investigate the distribution of single nucleotide polymorphisms(SNPs) on retinol binding protein 4(RBP4) genes and forkhead box O1 (FOXO1) gene, and their relationships with the occurrence of type Ⅱ diabetes mellitus (T2DM) in Chinese Han population. Methods Totally ten SNPs on RBP4 and FOXO1 were determined in 384 T2DM patients and 384 normal controls by TaqMan probe genotyping and agarose gel electrophoresis methods. And their serum level of fasting blood glucose (FBG), total cholesterol (TC) and trigly- ceride (TG) were also estimated. Results For RBP4, there was no significance for various genetypes and alleles including - 803 G > A, + 5169 C > T, and + 6969 G > C between two groups (P > 0.05). Each genotype had no relationships with T2DM (using adjusted logistic regression models). No haplotype was associated with T2DM. For FOXO1, among seven SNPs typed, significant variation was found in the frequency distribution of rs7324943 G/T in the two groups(χ~2=4.02, P = 0.044), and further stratification analysis showed that in subjects of aged 40 and non-hypertension, there was a higher risk of T2DM in GT heterozygous carriers than in GG homozygous carriers (OR = 1.47, 1.80), T allele carriers showed higher risk than non-T carriers (OR = 1.42,1.79). For rs17592236 C/T, though no significant frequency variation was found between two groups (χ~2 = 0.39, P = 0.401), but in subjects of aged ≤ 40, stratification analysis showed dramatically increased risk of T2DM in CT and TT carriers than in CC carriers (OR = 6.33,10.15), T allele carriers showed 7. 11-fold higher risk than non-T carriers. A haplotype CT related to T2DM susceptibility was also found, which could decrease the risk of its carriers by 28%. Conclusions For BBP4, the polymorphisms of - 803 G > A, + 5169 C > T, and + 6969 G > C had no relationships with T2DM in Chinese Han population. For FOXO1, the polymorphism of rs7324943 G/T,rs17592236 C/T and a haplotype CT were found related to the susceptibility of T2DM in Chinese Han population. Yet further studies are necessary to explain the impact of these polymorphisms on the disease occurrence.
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Objective To investigate the seruln levels of solubl B7-H3(sB7-H3)in patients withbacteremia and in those with chronic hepatitis B(CHB),and its clinical significance.Methods Serumlevels of sB7-H3 from 50 bacteremia patients,78 CHB patients and 50 healthy individuals wero detected byEUSA,and its correlations with the quantities of leucocytes,neutrophils,lymphocytes and monocytes wereanalyzed.Results The average level of sB7-H3 in serum of bacteremia patients was(50.69±26.43)μg/ml which was higher than that of healthy group(P<0.01);serum sB7-H3 level of patients with CHB was(18.07±7.28)μg/ml,and it had no significant difference compared with that in the healthy group(P>0.05).Besides,positive correlations between the scrum sB7-H3 level in and the quantities of leucocytesand granulocytes were observed in bacteremia patients.Conclusions It suggests that sB7-H3 misht bepotentially used as a new parameter to improve the clinical diagnosis of bacteremia.
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Objective To identify new serum biomarkers of lung adenocarcinoma.Methods Serum samples from 31 patients with lung adenocarcinoma and 31 healthy individuals were applied to SAX-2 protein chips to generate proteomic spectra by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry(SELDI-TOF-MS).The spectra were analyzed with Ciphergen Biosystems software and biomarker patterns software.Results The software identified 102 peaks and m/z 14 022.9 and 3 735.99 were used to construct the classification tree.The classification tree separated adenocarcinoma of lung effectively from healthy individuals,achieving a validity of 100%.The blind test challenged the model with a sensitivity of 100%and a specificity of 100%.Conclusions The results suggest that SELDI-TOF-MS technique can distinguish lung adenocarcinoma patients from healthy individuals and shows great potential for the development of a screening test for the detection of lung cancer.
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BACKGROUND: The angiotensin-converting enzyme (ACE) is the important component of the renin-angiotensin-aldosterone system (RAS). The ACE gene has, in fact,insertion/deletion polymorphism in intron 16, consisting of a 287-base pair Alu repeat sequence. ACE gene heterozygotes insertion/deletion (I/D) polymorphism is correlated with cardiovascular disease and IgA nephropathy and other diseases. OBJECTIVE: To investigate the distribution of ACE gene I/D polymor-phism in Chinese Han population in comparison with other known ethnic populations. DESIGN: Observation study on healthy individuals of Han nationality. SETTING: Key Laboratory of Clinical Immunology of Jiangsu Province; Department of Laboratory Medicine, First Hospital Affiliated to Soochow University; Department of aboratory Medicine, College of Medical Technology of Jiangsu University PARTICIPANTS: Totally 241 healthy individuals who received the healthy examination in the First Hospital of Soochow University between December 2005 and January 2006 were recruited in the experiment. They were 152 male and 89 female , with mean age of (27±8)years. All the participants without blood relationship were Han nationality from Suzhou region in China, free from disorder of hepatic, renal, endocrine and cardio- cerebrovascular diseases which were confirmed by clinical and experimen- tal examination. METHODS: Genotype of ACE gene I/D polymorphism allele of 241 healthy individuals of Han nationality was detected with polymerase chain reaction (PCR). PCR purified products with genotype of deletion/deletion (DD) and insertion/insertion (Ⅱ) polymorphism were performed DNA sequencing with fluorescence-labeled end termination method. MAIN OUTCOME MEASURES: Genotype and allele frequency of ACE gene I/D, as well as the comparison between them and those of other ethnic population. RESULTS: All the 241 subjects participated in final result analysis. ① The genotypes of ACE were DD, Ⅱ and ID. Compared with allele Ⅰ, allele D lost 287-base pair Alu repeat sequence. ②The frequencies of genotype Ⅱ, ID and DD were 46.1%, 41.5% and 12.4% respectively, with an allelic frequency of 66.8% for allele Ⅰ and 33.2% for allele D. ③The distribution of ACE genotype was similar between Japanese and Han nationality crowd, both presenting that type Ⅱ was commonly seen and type DD was the least; ID was mostly found in European and American crowd, but Ⅱ was little found. There was racial diversify of frequency of the distribution of ACE genotype among individuals of Han nationality and Japanese as well as Europeans and Americans. Compared with other nationalities, allele Ⅰ of individuals of Han nationality was significantly higher than that of above nationalities (χ2=105.55,P < 0.01), but allele D was obviously lower (χ2=87.54,P < 0.01). CONCLUSION: ACE gene polymorphism has racial diversify. To know genetic features of ACE gene polymorphism of individuals of different na tionalities is the basis and prerequisite to study the correlation of ACE gene I/D polymorphism with diseases.
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Objective To investigate the changes of the forkhead box transcription factor-1(FOX01) mRNA level in peripheral blood mononuclear cells(PBMC) from type 2 diabetes mellitus(T2DM) patients and to explore the role of FOX01 in pathogenesis of T2DM.Methods PBMC was isolated from 62 T2DM patients and 40 healthy persons.FOX01 mRNA level in PBMC was measured with reverse transcription PCR and real-time fluorescent quantitative PCR.Results FOX01 mRNA level in PBMC from T2DM patients was significantly higher than that from healthy persons(P
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Objective To evaluate the sensitivity and speciality of protein chip technology, and discuss its value in diagnosis or classification of autoimmune diseases, and to make its methodological evaluation. Methods The anti-dsDNA was detected with gold-colloid assay, indirect immunoflurescence(IIF) assay and protein chip technology, respectively; the other seven autoantibodies including anti-SSA, anti-SSB, anti-Sm, anti-u1RNP, anti-Rib-P, anti-Scl-70 and anti-Jo-1 were simultaneously detected with immunoblotting(IBT) assay and protein chip technology, and then all the results were delt with statistical method. Results For anti-dsDNA, the sensitivity of protein chip technology was better than that of gold-colloid assay; there was significant difference between protein chip technology and IBT assay in detecting anti-Jo-1 in DM/PM(P