flow cytometry

摘要： Acute lymphoblastic leukemia/lymphoma is a highly aggressive neoplasm of precursor lymphoid (blast) cells. There are 2 main subtypes based on lymphoid lineage;B lymphoblastic leukemia/lymphoma (B-ALL/LBL) and T lymphoblastic leukemia/lymphoma (T-ALL/LBL). B-ALL/LBL commonly presents with fever, fatigue, bone or joint pain, bleeding or anorexia (signs of bone marrow infiltration), lymphadenopathy, hepatosplenomegaly, involvement of skin, soft tissue and testes, with a predilection for the central nervous system. Immature cell markers, such as CD34 and TdT, can help to differentiate lymphoblasts from Burkitt lymphoma which, is considered a mature high-grade B cell lymphoma that mimics lymphoblastic lymphoma/leukemia. Unfavorable prognostic factors include: infancy and adult age of diagnosis, high white blood cell count, slow response to initial therapy, central nervous system involvement at the time of diagnosis and Minimal residual disease after therapy. We present a case report of a 4 months old infant seen at a Tertiary Hospital with a rare presentation of CD34 Negative B-lymphoblastic leukemia/lymphoma presenting as cutaneous lesions in infancy.

摘要： Background:Genetic selection in cattle has been directed to increase milk production.This,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have led to a reduction of fertility rates over the years.Thus,seeking sensitive and specific sperm biomarkers able to predict fertility rates is of vital importance to improve cattle reproductive efficiency.In humans,sperm chromatin condensation evaluated through chromomycin A3(CMA3)has recently been purported to be a powerful biomarker for sperm functional status and male infertility.The objectives of the present study were:a)to set up a flow cytometry method for simultaneously evaluating chromatin condensation and sperm viability,and b)to test whether this parameter could be used as a predictor of in vivo fertility in bulls.The study included pools of three independent cryopreserved ejaculates per bull from 25 Holstein males.Reproductive outcomes of each sire were determined by non-return rates,which were used to classify bulls into two groups(highly fertile and subfertile).Results:Chromatin condensation status of bovine sperm was evaluated through the combination of CMA3 and Yo-Pro-1 staining and flow cytometry.Sperm quality parameters(morphology,viability,total and progressive motility)were also assessed.Pearson correlation coefficients and ROC curves were calculated to assess their capacity to predict in vivo fertility.Sperm morphology,viability and total motility presented an area under the ROC curve(AUC)of 0.54,0.64 and 0.68,respectively(P>0.05),and thus were not able to discriminate between fertile and subfertile individuals.Alternatively,while the percentage of progressively motile sperm showed a significant predictive value,with an AUC of 0.73(P=0.05),CMA3/Yo-Pro-1 staining even depicted superior results for the prediction of in vivo fertility in bulls.Specifically,the percentage of viable sperm with poor chromatin condensation showed better accuracy and precision to predict in vivo fertility,with an AUC of 0.78(P=0.02).Conclusions:Chromatin condensation evaluated through CMA3/Yo-Pro-1 and flow cytometry is defined here as a more powerful tool than conventional sperm parameters to predict bull in vivo fertility,with a potential ability to maximising the efficiency of dairy breeding industry.

摘要： A wild rice (BKK) strain showing sterile spikelet and big leaves inhabited at the basin of the Chao Phraya river of Bangkok city, Thailand. The BKK strain was found as a natural triploid and thus its origin has been interested long time. Three different-sized fragments were amplified in RNA polymerase I largest subunit (PolA1) gene, which is a single-copy nuclear gene per haploid genome. Short type (0.14 kb) intron 20 sequence of BKK strain was identical to that of O. rufipogon and O. sativa. Phylogenetic analysis showed that long type (1.5 kb and 1.8 kb) intron 20 sequences of BKK strain were closely related to that of O. longistaminata and O. officinalis, respectively. We analyzed protein tag (Ptag) sequence encoded by exons 19 to 21 of PolA1 gene. Determined three Ptag sequences of BKK strain were identical to that of O. rufipogon, O. longistaminata, and O. officinalis, respectively. Relative DNA content of nuclei in O. officinalis and BKK strain was 1.5 and 1.75 times than that in O. sativa, respectively. And BKK strain contained CentO-C1 repeats, which were unique to O. officinalis. These results indicated that BKK strain comprised three genomes of O. rufipogon, O. longistaminata, and O. officinalis.

摘要： Erianthus arundinaceus is a wild relative of sugarcane(Saccharum officinarum L.)with many desirable agronomic traits for sugarcane genetic improvement.However,limited knowledge of the complex genome of hexaploid E.arundinaceus has impeded the development of required molecular tools.Dissecting complex genomes into single chromosomes can simplify analyses.The flow-cytometric sorting of a single chromosome of E.arundinaceus in a Saccharum-Erianthus introgression line is reported.A novel approach called genomic in situ hybridization in suspension was used to discriminate the alien chromosome from sugarcane chromosomes at the same size.A total of 218,000 E.arundinaceus chromosome 1(EaC1)were sorted to>97%purity and amplified DNA was sequenced using Illumina and Pac Bio technologies.The resulting assembly had a 70.93 Mb contig sequence with an N50 of 19.62 kb.A total of 56.69 Mb repeat sequences were predicted,accounting for 79.1%of the chromosome and 2646 genes having a total length of 1.84 Mb that represented 2.59%of the chromosome.Of these genes,1877(70.9%)genes were functionally annotated.The phylogenetic relationship of E.arundinaceus with other species using the chromosome1 sequence revealed that E.arundinaceus was distantly related to Oryza sativa and Zea mays,followed by Sorghum bicolor,and was closely related to S.spontaneum and Saccharum spp.hybrids.This study provides the first insights into the characteristics of EaC1,and the results will provide tools to support molecular improvement and alien introgression breeding of sugarcane.

摘要： [Objectives]This study was conducted to establish a method for detecting the immunophenotypes of venous blood CIK cells in healthy donors and patients with triple-negative breast cancer or lung cancer by flow cytometry,and to further analyze and discuss the proportion of cellular immunophenotypes such as CD3^(+),CD4^(+),CD8^(+),CD3^(+)CD8^(+),CD3^(+)CD56^(+)in CIK cells.[Methods]Human venous blood was drawn,then anticoagulated with heparin and isolated with lymphocyte isolation solution,and the relevant immunophenotypes were detected by flow cytometry after 21 d of culture.[Results]The expression of CD3^(+)CD8^(+)and CD3^(+)CD56^(+)in the venous blood CIK cells was significantly higher in healthy donors than that in triple-negative breast and lung cancer patients.[Conclusions]CD3^(+)CD8^(+)and CD3^(+)CD56^(+)CIK cells have high anti-tumor activity.

摘要： BACKGROUND Acute respiratory distress syndrome(ARDS)is injury of alveolar epithelial cells and capillary endothelial cells caused by various factors,including endogenous and exogenous lung factors,leading to diffuse pulmonary interstitial and alveolar edema,and acute respiratory failure.ARDS involves alveolar epithelial cells and pulmonary interstitial capillary endothelial cells.Circulating endothelial cells(CECs)are the only marker that directly reflects vascular endothelial injury in vivo.There have been few studies on the correlation between peripheral blood CECs and ARDS at home and abroad.The lungs are the organs with the highest capillary density and the most endothelial cells,thus,it is speculated that when ARDS occurs,CECs are stimulated and damaged,and released into the circulatory system.AIM To explore the correlation between CEC level and severity of ARDS in patients postoperatively.METHODS Blood samples were collected from all patients on day 2(d2)and day 5(d5)after surgery.The control group comprised 32 healthy volunteers.Number of CECs was measured by flow cytometry,and operation time was recorded.Changes in various indexes of patients were monitored,and diagnosis of ARDS was determined based on ARDS Berlin definition.We comprised d2 CECs in different groups,correlation between operation time and d2 CECs,ARDS of different severity by d2 CECs,and predictive value of d2 CECs for ARDS in postoperative patients.RESULTS The number of d2 CECs in the ARDS group was significantly higher than that in the healthy control group(P1351/mL,sensitivity and specificity of predicting ARDS were 80.8%and 78.1%,respectively.CONCLUSION Changes in number of CECs might predict occurrence and adverse outcome of ARDS after surgery,and higher numbers of CECs indicate worse prognosis of ARDS.

摘要： Flow cytometry is widely used for lymphocyte immunophenotyping in clinical settings.However,few studies have applied it for analyzing lymphocytes of the gastric mucosa.This review offers an overview of methodologies for isolating lymphocytes from the human stomach.Previously reported articles were reviewed,focusing on procedures for isolating human gastric mucosal lymphocytes.Helicobacter pylori-associated peptic diseases and gastric cancer are two major subjects of research in this field.Enzymatic dissociation,mechanical dissociation,or a combination of the two have been used to isolate lymphocytes from the stomach.Intra-epithelial and lamina propria lymphocytes were separately isolated in several studies.We also summarize the history and present trends in analyzing lymphocytes in patients with gastric disease.

摘要： AIM To examine the effect of high doses of vitamin C(VitC)on ex vivo human platelets(PLTs).METHODS Platelet concentrates collected for therapeutic or prophylactic transfusions were exposed to:(1)normal saline(control);(2)0.3 mmol/L VitC(Lo VitC);or(3)3 mmol/L VitC(Hi VitC,final concentrations)and stored appropriately.The VitC additive was preservative-free buffered ascorbic acid in water,pH 5.5 to 7.0,adjusted with sodium bicarbonate and sodium hydroxide.The doses of VitC used here correspond to plasma VitC levels reported in recently completed clinical trials.Prior to supplementation,a baseline sample was collected for analysis.PLTs were sampled again on days 2,5 and 8 and assayed for changes in PLT function by:Thromboelastography(TEG),for changes in viscoelastic properties;aggregometry,for PLT aggregation and adenosine triphosphate(ATP)secretion in response to collagen or adenosine diphosphate(ADP);and flow cytometry,for changes in expression of CD-31,CD41a,CD62p and CD63.In addition,PLT intracellular VitC content was measured using a fluorimetric assay for ascorbic acid and PLT poor plasma was used for plasma coagulation tests[prothrombin time(PT),partial thrombplastin time(PTT),functional fibrinogen]and Lipidomics analysis(UPLC ESI-MS/MS).RESULTS VitC supplementation significantly increased PLTs intracellular ascorbic acid levels from 1.2 mmol/L at baseline to 3.2 mmol/L(Lo VitC)and 15.7 mmol/L(Hi VitC,P0.05).PLT function assayed by TEG,aggregometry and flow cytometry was not significantly altered by Lo or Hi VitC for up to 5 d.However,PLTs exposed to 3 mmol/L VitC for 8 d demonstrated significantly increased R and K times by TEG and a decrease in theα-angle(P0.05).Collagen and ADP-induced ATP secretion was also not different between the three groups(P>0.05).Finally,VitC at the higher dose(3 mmol/L)also induced the release of several eicosanoids including thromboxane B2 and prostaglandin E2,as well as products of arachidonic acid metabolism via the lipoxygenases pathway such as 11-/12-/15-hydroxyicosatetraenoic acid(P<0.05).CONCLUSION Alterations in PLT function by exposure to 3 mmol/L VitC for 8 d suggest that caution should be exerted with prolonged use of intravenous high dose VitC.