Selective Cytotoxic effect of Probiotic, Paraprobiotic and Postbiotics of L. casei strains against Colorectal Cancer Cells: Invitro studies

Abstract This study highlights the cytotoxic effect of three L. casei strains on colorectal cell lines in invitro conditions. Different concentrations of live, heat killed (HK) and cell free supernatant (CFS) of three L.casei strains were subjected to CaCo2 and MRC5 cell lines. The viability of the treated and untreated cells was determined after 72 hrs by MTT assay, and IC50 estimated. Apoptosis was evaluated by Annexin V-propidium iodide method using flow cytometry. The live, HK and CFS of the L. casei strains showed cytotoxic effects on colorectal cell lines with significant differences. The cytotoxicity effects of live cells on CaCo2 cells were significantly higher (p˂0.01) than the HK cells. A dose dependent response was observed, as higher concentrations resulted in enhanced cytotoxicity effects. Live L.casei 1296-2cells inhibited 91% of CaCo2 cell growth, with IC50 of less than 108 cfu/ml. MRS medium and concentrations of CFS at above 20% v/v, were cytotoxic to the normal cell lines. Flow cytometry analyses of L. casei 1296-2 indicated that cytotoxicity effects on CaCo2 cells is related to apoptotic induction. Invitro studies indicate that Live and CFS of L. casei 1296-2 might be promising candidate for the control of colorectal cancers

Peripheral blood differential count of white blood cells in blood donor / Монголын Анагаах Ухаан

Background@#Establishment of quantitative reference intervals of white blood cells and its subpopulations using a high accuracy analytic system is essential for clinical medicine, public health, and anthropology. We are unable to identify peer-reviewed literature sources describing white blood cell counts and their subpopulations using monoclonal antibodies to specific surface antigens in healthy Mongolians. This study aimed to measure the counts of white blood cells and their subpopulations in healthy Mongolians using flowcytometry. @*Materials and Methods@#The absolute number (cell/L) of leukocytes (CD45+), granulocytes, monocytes and lymphocytes were measured by Magnetic Activated Cell Sorting Assay (MACSQuant Analyzer 10) in 287 blood donors (158 males and 129 females) 17-64 years of age (mean age 33.1±12.4). Peripheral blood samples were collected at the time of blood donation at the National Center for Transfusion Medicine.@*Results@#The mean values of leukocytes and granulocytes were lower in donors over 30 years of age (ANOVA: F=4.408, p=0.002 and F=5.685, p=0.001) and regression analysis demonstrated indirect correlation between counts of these cells and age of donors (r= - 0.198, p=0.001 and r=-0.221, p=0.001, respectively). Gender-related differences in white blood cell counts were not found.</br> Mean value of lymphocyte count in donors investigated in spring (May and March, n = 87; 2224.6±775.3) was significantly higher than those in winter (December – February, n=180; 1613.2±454.3, p=0.001) and autumn (October, n=20; 1576.1±438.6, p= 0.001). </br> Comparing of our findings with the data from available literature shown that healthy Mongolians have lower leukocyte count compared with Koreans, Chinese Han population and lower mean value of lymphocyte count comparing with Korean, Chinese Han population, and Arabian (Saudi Arabia) populations.

To Assess the Correlation between Bone Marrow Morphology and Immunophenotypic Findings in Patients of Chronic Lymphoproliferative Disorders (CLPD’s) and to Assess the Role of Flowcytometric Immunophenotyping in Diagnosis and Subclassification of CLPD’s

Introduction: Chronic lymphoproliferative disorderrepresent clonal proliferation of morphologically andimmunophenotypically mature B or T cells characterized by alow proliferation rate and prolonged cell survival. Study aimedto assess the correlation between bone marrow morphologyand immunophenotypic findings in patients of ChronicLymphoproliferative Disorders (CLPD’s) and to assess therole of flowcytometric immunophenotyping in diagnosis andsubclassification of CLPD’s.Material and Methods: 48 newly diagnosed cases ofCLPD were included. After complete clinical evaluation theyunderwent marrow aspiration, biopsy and immunophenotypingby flowcytometry with selected panel of monoclonalantibodies.Results: On morphology 47.9% cases were CLL. In 52.1%non CLL cases , 4.2% were PLL , 2% case as LPL and45.8% cases were CLPD-unclassifiable. Commonest patternof marrow infiltration noted on trephine biopsy was diffuse inCLL, HCL-V, B-PLL and T-CLPD. On immunophenotyping95.8% cases were B-CLPD and 4.25% T-CLPD. CD5, CD22,CD23, FMC7 and SmIg were used as first line markersfollowed by CD 10, CD 25, CD103, CD38, CD138 andCyclin D1 (on biopsy sections) as second line markers. Finalimmunophenotypic diagnosis was CLL (54.2%), B-CLPDunclassified (29.2%), 4.1% each of LPL, MCL, T-CLPD and2% each of B-PLLand HCL-V.Conclusion: Concordance rate between morphologicaldiagnosis and immunophenotypic diagnosis was 79.17%.Hence, Flowcytometry is necessary for confirmationof diagnosis and to classify the CLPD cases which areunclassifiable by morphology

Correlation of Cytomorphology with Flowcytometric Immunophenotyping of Acute Myeloid Leukemia in Tertiary Care Hospital

Introduction: WHO classification utilizes morphology,genetic information, immunophenotyping, biologic andclinical features to define specific disease entity. Althoughit gives an accurate detailed diagnosis, immunophenotypingby flow cytometry gives an immediate prompt diagnosis.Morphological diagnosis for leukemias may sometimes be notcorrelating with flow cytometry diagnosis. Study objectiveswere to correlate morphological and flowcytometric results ofpatients diagnosed with acute myeloid leukemias.Material and methods: Study was conducted in departmentof pathology. Cases were classified as Acute leukemia basedon CBC, peripheral smear, bone marrow morphology, specialstains cytochemistry and Flow cytometry Immunophenotyping.Categorization was done based FAB system.Results: Total 92 cases of AML were diagnosed oncytomorphology, cytochemistry and Flow cytometry werestudied. Out of which M0 were 6.5%, M1-13%, M2-27.2%,M3-17.4%, M4-15.2% and M5 were 20.6%. There was 88%correlation between cytomorphology and flowcytometry.Conclusion: Interpretation of immunophenotyping byflowcytometry, done in close conjunction with morphology,is mandatory for appropriate diagnosis of acute myeloidleukemia. However morphology combined with cytochemistryis also very helpful in the diagnosis of AML if facility offlowcytometry is not available

Analysis of poliovirus receptor, CD155 expression in different human colorectal cancer cell lines: Implications for poliovirus virotherapy

Context: Poliovirus (PV) receptor (CD155) is expressed on several kinds of cells and exerts diverse functions. Various investigations have confirmed that changes in CD155 expression in cancer cell lines affect metastasis, proliferation, and migration. Aims: The purpose of the present study was to investigate the CD155 transcript and protein expression in human colon adenocarcinoma cell lines in comparison to normal fetal human colon (FHC) cells. Materials and Methods: The CD155 expression level in four human adenocarcinoma cell lines and normal colon cell line were assessed using the SYBR green quantitative real-time polymerase chain reaction (PCR) and flowcytometry. Results: The results of real-time PCR indicated that CD155 was significantly overexpressed in all human adenocarcinoma cell lines (P = 0.000). The highest and the lowest expression level of CD155 messenger RNA was observed in SW480 and HT29 cell lines by 491.14, and 12.04 fold changes, respectively, in comparison with the human normal cell line (FHC). Results of flowcytometry indicate that protein was strongly expressed in cancer cell lines. SW480 cells showed the highest CD155 protein expression level of 98.1%, whereas this protein expression was 1.3% in human normal colon cell line (FHC). Totally, these data indicate that CD155 expression is significantly elevated in cancer cell lines. Conclusions: The preferential expression of CD155 on cancer cell lines rather than on normal cell line suggests that CD155 could be targeted for future PV virotherapy

ExperimentalStudyonDetectionofDeadandLive Mycobacterium Tuberculosis by FlowCytometry / 中山大学学报(医学科学版)

@#：【Objective】To provide basic experimental basis for quantitative detection and analysis of antibiotic susceptibilityandheteroresistancein Mycobacterium tuberculosis byflowcytometry，weinvestigatedtheperformanceof propidiumiodide（PI）and6-carboxyfluoresceindiacetae（6-cFDA）inidentifyingdeadorlivingMycobacterium tubercu⁃ losis.【Methods】PIand6-cFDAwereusedtostain10strainsofpureculturedliving Mycobacterium tuberculosis suspen⁃ sion with turbidity of 1 McClell and the corresponding suspension with turbidity of 1 McClell heated at 85℃ for 30 minutes，respectively.Themeanfluorescenceintensity（MFI）wasmeasuredbyflowcytometry.TheMFIboundaryvalue fordistinguishingdeadandlivingMycobacterium tuberculosisweredeterminedaccordingtoreceiveroperatingcharacteristic （ROC）curves.Thesameexperimentswerecarriedoutwithother10strainsofliving Mycobacterium tuberculosis suspen⁃sionand10dead Mycobacterium tuberculosis suspension，andtheMFIthresholdwasusedtodeterminetheviabilityof bacteria.【Results】（1）IntheexperimentofdeterminingMFIboundaryvalue，theMFIof10strainsofdeadbacteria suspensionstainedwithPIwasnon-normaldistribution，themedianvaluewas2459，andtheMFIof10strainsofliving bacteriasuspensionstainedwithPIwasnormaldistribution，themeanvaluewas426±180.TheMFIboundaryvalueof deadbacteriastainedwithPIwas1329.TheMFIof10strainsofdeadbacteriaand10strainsoflivingbacteriasuspen⁃ sionstainedwith6-cFDAwasnormaldistribution，andthemeanvaluewas49±4and7144±4511，respectively.The MFIthresholdof6-cFDAstainingforlivingbacteriawas1021.（2）TheMFIthresholdsforidentifyingdeadbacteriaby PIstainingandtheMFIthresholdsforidentifyinglivingbacteriaby6-cFDAstainingwereusedtodetect10otherliving bacterialsuspensionsand10deadbacterialsuspensions.Theaccuracy，sensitivity，specificity，Yondeindex，positive predictivevalueandnegativepredictivevaluewere95%and100%，1.00and1.00，0.90and1.00，0.90and1.00，0.91 and1.00，1.00and1.00，respectively.【Conclusions】PIstainingcandetectdeadMycobacterium tuberculosisand6-cFDA stainingcandetectliving Mycobacterium tuberculosis.Identificationof Mycobacterium tuberculosis activitybasedonPIand 6-cFDAstainingwillhavebroadapplicationprospects.

Bernard Soulier Syndrome; A Rare Bleeding Disorder

Bleeding syndromes in the newborn are rare, but they may be life-threatening and demand immediate attention. Congenital bleeding disorders especially pose a diagnostic challenge to the clinician because of their rarity and the need to be differentiated from the other common causes of bleeding in children. We present a case of an infant presenting with bleeding symptoms early in his life (since 5 months of age) which was initially thought to be immune thrombocytopenic purpura (ITP) with low platelet count. No response to steroids and further evaluation by platelet aggregometry and flowcytometry led to the correct diagnosis – Bernard soulier syndrome(BSS). Though, there is no specific treatment available for this rare bleeding disorder, however it is imperative to have arrived at correct diagnosis in order to save unnecessary therapy and to take due precautions for prevention of bleeding.

Bernard Soulier Syndrome; A Rare Bleeding Disorder

Bleeding syndromes in the newborn are rare, but they may be life-threatening and demand immediate attention. Congenital bleeding disorders especially pose a diagnostic challenge to the clinician because of their rarity and the need to be differentiated from the other common causes of bleeding in children. We present a case of an infant presenting with bleeding symptoms early in his life (since 5 months of age) which was initially thought to be immune thrombocytopenic purpura (ITP) with low platelet count. No response to steroids and further evaluation by platelet aggregometry and flowcytometry led to the correct diagnosis – Bernard soulier syndrome(BSS). Though, there is no specific treatment available for this rare bleeding disorder, however it is imperative to have arrived at correct diagnosis in order to save unnecessary therapy and to take due precautions for prevention of bleeding.

Comparison of the Tellgenplex HPV DNA test with the PCR-reverse dot blot assay for human papillomavirus genotyping

Objective:To access the performance of the Tellgenplex human papillomavirus (HPV) DNA test compared to the polymerase chain reaction-reverse dot blot (PCR-RDB) assay for the HPV genotyping.Methods:Sixty cervical swab samples were genotyped by the Tellgenplex HPV DNA test and the PCR-RDB assay. The Tellgenplex HPV DNA test and the PCR-RDB assay can detect 26 and 23 HPV genotypes, respectively. Each sample showed discrepancy was genotyped using sequencing.Results:The percent agreement between the two tests ranged from 83.3% to 100.0% according to different genotype. This showed perfect agreement (>0.81) for high-risk HPV genotypes (35, 39, 45, 53, 56, 59, 66, 68, and 82), substantial agreement (>0.65) for high-risk HPV genotypes (16, 18, 33, 52, and 58) and low-risk HPV genotype 43 between the two assays by the kappa analysis. The positive rates of the two assays for frequent HPV genotypes (16, 35, 39, 45, 52, 53, 58, 59, 66, and 82) were not statistically different, but the PCR-RDB assay showed higher positive rates than the Tellgenplex HPV DNA test for HPV genotypes 81 (P<0.05). As for more than 10 positive results by the Tellgenplex HPV DNA test and/or the PCR-RDB assay, the PCR-RDB assay showed higher relative sensitivity and specificity than the Tellgenplex HPV DNA test for the three HPV genotypes (16, 52, and 81). All HPV genotypes that can be detected by only the Tellgenplex HPV DNA test (HPV genotypes 44 and 55) were confirmed by sequencing.Conclusions:In conclusion, our results demonstrated that the PCR-RDB assay which can detect more multiple HPV genotypes in each specimen shows higher relative sensitivity and specificity than the Tellgenplex HPV DNA test, which makes it a better option for routine clinical use.

Comparison of the Tellgenplex HPV DNA test with the PCR-reverse dot blot assay for human papillomavirus genotyping

Objective: To access the performance of the Tellgenplex human papillomavirus (HPV) DNA test compared to the polymerase chain reaction-reverse dot blot (PCR-RDB) assay for the HPV genotyping. Methods: Sixty cervical swab samples were genotyped by the Tellgenplex HPV DNA test and the PCR-RDB assay. The Tellgenplex HPV DNA test and the PCR-RDB assay can detect 26 and 23 HPV genotypes, respectively. Each sample showed discrepancy was genotyped using sequencing. Results: The percent agreement between the two tests ranged from 83.3% to 100.0% according to different genotype. This showed perfect agreement (>0.81) for high-risk HPV genotypes (35, 39, 45, 53, 56, 59, 66, 68, and 82), substantial agreement (>0.65) for high-risk HPV genotypes (16, 18, 33, 52, and 58) and low-risk HPV genotype 43 between the two assays by the kappa analysis. The positive rates of the two assays for frequent HPV genotypes (16, 35, 39, 45, 52, 53, 58, 59, 66, and 82) were not statistically different, but the PCR-RDB assay showed higher positive rates than the Tellgenplex HPV DNA test for HPV genotypes 81 (P<0.05). As for more than 10 positive results by the Tellgenplex HPV DNA test and/or the PCR-RDB assay, the PCR-RDB assay showed higher relative sensitivity and specificity than the Tellgenplex HPV DNA test for the three HPV genotypes (16, 52, and 81). All HPV genotypes that can be detected by only the Tellgenplex HPV DNA test (HPV genotypes 44 and 55) were confirmed by sequencing. Conclusions: In conclusion, our results demonstrated that the PCR-RDB assay which can detect more multiple HPV genotypes in each specimen shows higher relative sensitivity and specificity than the Tellgenplex HPV DNA test, which makes it a better option for routine clinical use.

Plasma-Derived Microparticles in Polycythaemia Vera

Introduction: Microparticles are membrane bound vesicles, measuring less than 1.0 um, which are released during cellular activation or during apoptosis. Studies have shown that these circulating microparticles play a role in coagulation, cell signaling and cellular interactions. Increased levels of circulating microparticles have been observed in a number of conditions where there is vascular dysfunction, thrombosis and inflammation. The objective of this study was to determine the various plasma-derived microparticles in patients with polycythaemia vera (PV) in Universiti Kebangsaan Malaysia Medical Centre and to compare them with normal control. Methods: A total of 15 patients with PV and 15 healthy volunteers were included in this cross-sectional descriptive study. Plasma samples from both patients and healthy volunteers were prepared and further processed for isolation of microparticles. Flow cytometry analyses were then carried out in all samples to determine the cellular origin of the microparticles. Full blood count parameters for both groups were also collected. Data collected were analyzed using SPSS version 12.0. Results: Patients with PV had a significantly higher percentage of platelet derived microparticles compared to healthy controls (P 0.05). Conclusion: The median percentage of positive events for platelet derived microparticles was higher in patients with PV compared to normal healthy controls.

New perspectives on the nutritional factors influencing growth rate of Candida albicans in diabetics. An in vitro study

BACKGROUND The link between Candida albicans and diabetes mellitus is well-acknowledged, but incompletely elucidated. OBJECTIVES The purpose of this study is to assess the growth rate of C. albicans (CA) in the presence of different concentrations of glucose and fructose, two of the main pathophysiologic and nutritionally relevant sugars in diabetic patients, in order to obtain a better understanding of the nutrient acquisition strategy and its possible relation to the hyperglycemic status of diabetic patients. METHODS The effects of different concentrations of glucose and fructose (1000 mg%, 500 mg%, 250 mg% and 100 mg% w/v) on the growth rate of CA have been studied by flow-cytometry. FINDINGS We found that glucose concentration is directly related to CA growth, which may be linked to the frequent yeast infections that occur in non-controlled diabetic patients; we also show that fructose inhibits CA growth rate. MAIN CONCLUSIONS As a consequence of our hypothesis, the study demonstrates that fructose-containing food may prevent the development of candidiasis, at least in oral sites.

Influence of bloody cervical cell samples on HPV genotyping detection / 国际检验医学杂志

Objective To investigate the influence of bloody cervical cell samples on human papillomavirus(HPV) genotyping detection using flowcytometry fluorescence hybridization kit.Methods According to the concentration of hemoglobin(Hb),cervical cell samples were divided into five groups,including group A with Hb of 0 g/L(10 cases),group B with 0 g/L100 g/L.In each group,50% samples were with initial positive results of HPV genotyping detection.Every sample was detected using sample volume of 50,100,200,500 and 1 000 μL different volumes for testing and analysis results.Results For all samples with initial negative results,five samples with initial positive results in group A and five samples with initial positive results,the detected results of different sample volume were without differences.In the 10 samples,with initial positive results of group C,D and E,there were four,four and nine samples respectively,were with false negative detected results of different sample volumes.Conclusion Bloody cervical cell samples could cause false negative results of HPV genotyping detection using flowcytometry fluorescence hybridization kit.It might be necessary to set optimal sample volume according to the concentration of Hb for ensuring the accuracy of genotyping detection.

High fms-like tyrosine kinase-3 (FLT3) receptor surface expression predicts poor outcome in FLT3 internal tandem duplication (ITD) negative patients in adult acute myeloid leukaemia: a prospective pilot study from India.

Background & objectives: Mutations in fms-like tyrosine kinase 3 (FLT3) receptor have significant role in assessing outcome in patients with acute myeloid leukaemia (AML). Data for FLT3 surface expression in relation to FLT3 internal tandem duplication (ITD) status and outcome are not available from India. The objective of the current study was to investigate adult patients with AML for FLT3 expression and FLT3 ITD mutation, and their association with long-term outcome. Methods: Total 51 consecutive de novo AML patients aged 18-60 yr were enrolled in the study. FLT3 ITD was detected by polymerase chain reaction (PCR); flowcytometry and qPCR (Taqman probe chemistry) were used for assessment of FLT3 protein and transcript, respectively. Kaplan Meier curves were obtained for survival analysis followed by log rank test. Results: FLT3 ITD was present in eight (16%) patients. Complete remission was achieved in 33 (64.6%) patients. At 57.3 months, event free survival (EFS) was 26.9±6.3 per cent, disease free survival (DFS) 52.0±9.2 per cent, and overall survival event (OS) 34.5±7.4 per cent. FLT3 surface expression was positive (>20%) by flow-cytometry in 38 (88%) of the 51 patients. FLT3 surface expression and transcripts were not associated with FLT3 ITD status. FLT3 expression was significantly associated with inferior EFS (P=0.026) and OS (P=0.018) in those who were negative for FLT3 ITD. Interpretation & conclusions: This study evaluated FLT3 ITD mutation along with FLT3 expression in AML patients, and associated with survival. Negative impact of FLT3 surface expression on survival was observed in AML patients who were FLT3 ITD negative.

Analysis of immunogenicity ofΔ42PD1 via yeast surface displaying peptide frag-ments / 中国免疫学杂志

Objective:To analyze the immunogenicity of the extracellular region of Δ42PD1.Methods: Six fragments ofΔ42PD1 extracellular region-encoding sequence were amplified by PCR, and were cloned into pCTCON2 vector, a yeast surface displaying vector.Yeast cells were transfected with Δ42PD1 fragment-carrying plasmids, then yeast cells were spread on SDCAA plates.Single cell clones were selected and cultured in SGCAA media to induce expression of the target genes.Mouse anti-humanΔ42PD1 anti-serum were generated by immunization of BALB/c mice via intramuscular injection ofΔ42PD1-carrying plasmid plus in-situ electroporation.The binding of anti-serum with yeast cells surface-displaying Δ42PD1 fragments were analyzed using flowcytometry.Results:Nucleotide sequences analysis indicated that the amplified six fragments ofΔ42PD1 sequence length were 110 bp,and the isolated sequence ofΔ42PD1 fragments were 100%homology with PD1 gene previously registered in GenBank.Results from flowcytometry showed that among the six fragments of Δ42PD1 displaying on the surface of yeast cells,F3 and F2 profoundly boundΔ42PD1-specific polyclonal antibodies.Conclusion:F3 and F2 ofΔ42PD1 is an immunogenic dominant region,which pave the way for generation of Δ42PD1-specific monoclonal antibody and epitope mapping.

Characterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1.

Viral gene oncotherapy, targeted killing of cancer cells by viral genes, is an emerging non-infectious therapeutic cancer treatment modality. Chemo and radiotherapy in cancer treatment is limited due to their genotoxic side effects on healthy cells and need of functional p53, which is mutated in most of the cancers. VP3 (apoptin) of chicken infectious anaemia (CIA) and NS1 (Non structural protein 1) of Canine Parvovirus-2 (CPV-2) have been proven to have oncolytic potential in our laboratory. To evaluate oncolytic potential of VP3 and NS1 together these genes needed to be cloned in a bicistronic vector. In this study, both these genes were cloned and characterized for expression of their gene products and its apoptotic potential. The expression of VP3 and NS1 was studied by confocal microscopy and flowcytometry. Expression of VP3 and NS1 in pVIVO.VP3.NS1 transfected HeLa cells in comparison to mock transfected cells indicated that the double gene construct expresses both the products. This was further confirmed by flowcytometry where there was increase in cells expressing VP3 and NS1 in pVIVO.VP3.NS1 transfected group in comparison with the mock control group. The apoptotic inducing potential of this characterized pVIVO.VP3.NS1 was evaluated in human cervical cancer cell line (HeLa) by DNA fragmentation assay, TUNEL assay and Hoechst staning. This double construct was observed to induce apoptosis in HeLa cells.

Correlation of aspirin resistance, gene polymorphism of platelet membrane glycoprotein Ⅱb HPA-3 and recurrent ischemic stroke / 国际脑血管病杂志

ObjectiveToinvestigatethecorrelationofaspirinresistance(AR),genepolymorphismof platelet membrane glycoprotein (GP)Ⅱb HPA-3 and recurrent ischemic stroke. Methods The consecutive patients w ith acute ischemic stroke and gender, age-matched healthy subjects w ere enrol ed. Adenosine diphosphate (ADP) and arachidonic acid (AA) w ere used as inducing agents. The platelet aggregation rate (PAgT) was detected by flow cytometry. AR was defined as PAgTADP≥39.27% and PAgTAA≥34.27%. Polymerase chain reaction-restriction fragment length polymorphism w as used to detect the GPⅡbHPA-3 genotype. Results A total of 224 patients w ith acute ischemic stroke (case group) and 98 healthy subjects (control group) were enroled. In the case group, 162 patients had the first-ever stroke (first-ever stroke group) and 62 had recurrent stroke (recurrent stroke group). The incidence of AR in the case group w as 15.18%, in w hich the incidence of AR in the recurrent group w as significantly higher than that in the first-ever stroke group (27.42%vs.10.49%; χ2 =9.977, P=0.002). The frequencies of bb genotype ( P=0.004) and b alele (P=0.001) in the recurrent group were significantly higher than those in the first-ever stroke group. In the case group, the frequencies of bb genotype ( P=0.028) and b alele (P=0.004) in the AR group w ere significantly higher than those in the non-AR group. Multivariable logistic regression analysis show ed that AR (odds ratio [OR] 2.933, 95%confidence interval [CI] 1.326-6.486;P=0.008) and bb genotype (OR 2.198, 95%CI1.164-4.149, P=0.015) w ere the independent risk factors for recurrent ischemic stroke. Conclusions AR and GP Ⅱb HPA-3 bb genotype are associated w ith recurrent ischemic stroke.

Value of flowcytometry fluorescence immunoassay of CA242 in the diagnosis of pancreatic cancer / 中华胰腺病杂志

Objective To conduct a methodological evaluation on flowcytomotry fluorescence detection of serum CA242, and to evaluate its value in the diagnosis of pancreatic cancer.Methods The blood samples of 40 cases of pancreatic cancer, 49 cases of other tumors, 48 cases of benign digestive diseases and 159 healthy volunteers were collected.Fowcytometry fluorescence immunoassay was used to detect serum level of CA242, and it was compared with routine ELISA method to measure its sensitivity and specificity.Results The detection limit of CA242 by flowcytometry fluorescence immunoassay was 0.89 U/ml, the linear range was 1～ 500 U/ml after confirmation.The within-batch CV was 3.37％～ 5.30％, between batch CV was 7.43％ ～ 9.60％.When compared with routine ELISA, flowcytometry fluorescence immunoassay showed the equation of linear regression is Y =1.0398X-0.947, r =0.9687.Area under ROC curve was 0.811 ± 0.025 (95％ CI 0.763 ～0.859), with 18.625 U/ml as the best cutoff value, the specificity was 92.0％ and the sensitivity was 62.1％.Conclusions Flowcytometry fluorescence immunoassay for CA242 testing has the advantage of shorter detection time, miltiple sample and testing project detection, which is worth of clinical application.

Changes of circulating Tc17 cells in progression of colorectal cancer / 中国免疫学杂志

Objective:To explore the changes and possible mechanisms of circulating Tc 17 cells in patients with colorectal cancer ( CRC) following disease progression .Methods:The peripheral blood were collected from 54 patients with colorectal cancer . the proportions of Tc17 cells in peripheral blood mononuclear cells (PBMCs) and cultured PBMCs treated by IL-1β, IL-6 and TGF-βin different concentrations were determined by flow cytometry;the levels of IL-1β, IL-17A, IL-23 and IL-6 in sera were measured by ELISA.Results:The proportions of Tc17 cells and the levels of IL-1β, IL-17A, IL-23 and IL-6 were significantly higher than those in healthy controls (P0.05).In vitro experiments confirmed that IL-1βor high concentrations of IL-6 and TGF-βcould significantly increase the number of Tc 17 cells in PBMCs.Conclusion: The changes of circulating Tc 17 cells in the progression of colorectal cancer are possibly modulated by IL-1β, IL-23, IL-6 and TGF-β.

Susceptibility tests of oropharyngeal Candida albicans from Egyptian patients to fluconazole determined by three methods

Candida albicans frequently cause oropharyngeal candidiasis in immunocompromised patients. As some of these isolates show resistance against azoles, the clinician is wary of initiating therapy with fluconazole (FZ) until a final susceptibility report is generated. We aimed to evaluate the efficacy of rapid flow cytometry (FCM) and disc diffusion (DD) methods in comparison to reference microdilution (MD) of Clinical and Laboratory Standards Institute (CLSI) method for FZ. Thirty seven Candida albicans isolates were tested by the three methods. By both MD and FCM, 26/37 (70.3%) were sensitive with minimal inhibitory concentration (MIC) ¡Ü 8¦Ìg/ml, 5/37 (13.5%) were susceptible dose dependant (S-DD) with MIC 16-32 ¦Ìg/ml and 6/37 (16.2%) were resistant with MIC ¡Ý64¦Ìg/ml. More than 92% of isolates susceptible to FZ by the MD were susceptible by the DD methods with good agreement (81.08%, P = 0.000). However, 4/5 isolates diagnosed as S-DD by MD were resistant by DD. Interestingly, the MIC by FCM at 4 h showed excellent agreement (95.59%, P = 0.000) to that obtained by MD method at 24 h. Overall, FCM antifungal susceptibility testing provided rapid, reproducible results that are valuable alternative to MD. The DD test is recommended as a simple and reliable screening test for the detection of susceptible Candida albicans isolates to FZ.