Impact of regulatory T-lymphocytes on therapeutic efficacy of human acute brucellosis / 中华传染病杂志

Objective To investigate the humoral and cellular immune responses in patients with acute brucellosis, and evaluate dynamic changes of regulatory T-lymphocytes (Foxp3+ Treg) in the peripheral blood of patients during treatment, in order to clarify the relationship between immunosuppression and the therapeutic effect in human brucellosis.Methods Sixty-five patients with brucellosis hospitalized at the Third Department of Infectious Diseases, Heilongjiang Agriculture and Reclamation Bureau General Hospital between July 2015 and November 2015 were included.Twenty-eight patients were treated with conventional therapy (group A: patients received 3 courses of treatment.Each lasted for 20 days with one-week interval), and 37 patients were treated with conventional therapy in combination with immunopotentiator (group B).Thirty healthy volunteers were enrolled as the controlled group.The ratio of CD3+CD4+ Foxp3+ Treg cells in the peripheral blood of brucellosis patients were measured by flow cytometry (FCM) at the end of each course of treatment.Data in accordance with normal distribution were described as mean±standard deviation.Comparison between two groups was done by two sample t test.Comparison among multiple groups was performed by analysis of variance and SNK test.Data that did not fit the normal distribution were analyzed by multiple-sample nonparametric test.Results After the first (20 d), second (50 d) and third course of treatment (80 d), the ratios of Foxp3+Treg in the peripheral blood of 65 acute brucellosis patients were 2.83%, 3.77% and 4.03%, respectively, which were all significantly higher than control group (1.69%;t=5.97, 9.05 and 5.66, respectively, all P0.05), while those were both higher than control group (t=7.09 and 4.94, respectively;both P<0.01).At the end of the second course, the ratio of Foxp3+ Treg in group B was higher than group A (t=2.22, P<0.01), and both of them were higher than control group (t=10.79 and 7.25, respectively;both P<0.01).At the end of treatment, Foxp3+ Treg in group A was also significantly higher than the other two groups (t=6.02 and 6.45, respectively;both P<0.01).Conclusions In patients with acute brucellosis treated with the standard antibiosis treatment in combination with immunopotentiator, the ratio of Foxp3+Tregs significantly increases and maintains at a high level, which suggests that extra immunopotentiator may be not helpful for the treatment of brucellosis at the very early stage.

Estimation of Genome Size of Ginseng Based on Herbgenomics by Flow Cytometric Analysis And High-throughput Sequence / 世界科学技术-中医药现代化

Ginseng is the dried root and rhizome of Panax ginseng.The lack of genomic data has restricted the development of ginseng industry and basic research.The genome size of P.ginseng was estimated to be 3.42 Gb by using the genome data of Oryza sativa ssp.Nipponbare and Glycine max (L.) Merrill as the reference and the flow cytometric analysis.Meanwhile,shotgun libraries with the insert size of 250 bp and 500 bp were constructed,and sequenced for double terminal PE 150 by using Illumina Hiseq X Ten platform.Totally,183.82 Gb high quality data was obtained after filtering the raw data.The genome size of P.ginseng was 3.35 Gb and the sequencing depth was 54.87 X by K-mer analysis.In this study,flow cytometry and K-mer analysis were used to identify the genome size of ginseng,which provided basic data for the further whole genome sequencing and herbgenomics studies.

Hairy-cell leukemia variant: a case report and review of the literatures / 白血病·淋巴瘤

Objective To study the clinical features and differential diagnosis of hairy-cell leukemia variant (HCL-V).Methods A case with HCL-V was reported and the literatures were reviewed.Results The patient had splenomegaly for twenty years and a history of recurrent pulmonary infection.His blood routine test showed a high white blood cell count and abnormal high proportion of lymphocytes.Peripheral smear and bone marrow smear both showed significantly higher proportion of lymphocytes,part of which had soma jagged,prominent nucleoli and villous/hairy cytoplasmic projections.His hairy leukemic cells expressed CD1 1c,CD19,CD20,CD22,and had variable expression of FMC-7,CD103 and lambda,but not CD5,CD23 and CD25.Transmission electron microscope showed many monocytes with villous exist in peripheral blood.Conclusions HCL-V is a rare and an indolent form of a small,mature,B-cell leukemia,based on the clinical,peripheral smear,bone marrow smear,flow cytometric analysis and transmission electron microscopy features,a diagnosis of HCL-V is confirmed.The differential diagnosis should always include splenic marginal zone B-cell lymphoma and HCL-C,because they have different clinical and biological features.

The Correlation between the Tc2 Cells and the CCR4 Expression and the PASI Score in the Peripheral Blood of Psoriasis Patients / 대한피부과학회지

BACKGROUND: Psoriasis is a chronic relapsing skin disorder that is characterized by abnormal epidermal proliferation, inflammation and angiogenesis. It causes emotional and social consequences that go far beyond the skin; therefore, many methods to measure and monitor the severity of psoriasis have been reported. OBJECTIVE: This study aims to evaluate the usability of the flow cytometric analysis of the T cell subsets and their chemokine receptors in the peripheral blood of the psoriasis patients as a severity index. METHODS: The T cell subsets and their chemokine receptor expression (CXCR3, CCR4) in the circulating blood of thirty psoriasis patients (PASI score:2.2~44.2) and twenty healthy controls were examined by flow cytometry. The relationship between the PASI score and the T cell subsets/chemokine receptors was also analyzed. RESULTS: The patients showed significantly higher number of Tc1 (CD8+CXCR3+), Tc2 (CD8+CCR4+) and CXCR3/CCR4 expressing cells than did the control group. Especially, the moderate to severe patients (a PASI score greater that 5) showed a higher number of Tc1, Tc2 and CCR4 expressing cells than did the control group. In the severe patients (a PASI score greater than 10), the frequency of circulating Tc2 cells and CCR4 expressing cells was directly correlated with the PASI score. CONCLUSION: Our findings suggest that flow cytometric analysis of the circulating T cell subsets with further classification could serve as an indicator of the disease severity in psoriasis patients.

An experimental study on mistletoe extract-induced apoptosis in oral squamous cell carcinoma

This study was performed to investigate mistletoe extract-induced apoptosis in oral squamous cell carcinoma. In vivo study, HN22 cells were xenografted in nude mice. After tumor was experimentally induced, mistletoe extract was directly injected on the tumor mass. The specimens were evaluated using light and transmission electron microscopes. In vitro study, HN22 cells were cultured and exposed to mistletoe extract. The cells were evaluated using transmissin electron microscope. To evaluate apoptotic cells, flow cytometric analysis was done. The results were obtained as follows: 1. Light microscopic view of tumor mass showed necrosis at 2-4 weeks. 2. Transmission electron micrographs of tumor mass showed apoptosis and necrosis. 3. In TEM view of cell lines, necrosis and apoptosis were shown with mistletoe extract at 300microgram/ml, apoptosis was shown with mistletoe extract at 100microgram/ml. 4. In flow cytometric analysis, early and late apoptosis was shown when using caspase-3Ab and annexin-V, but no significant change was noted when using mebstain and Apo2.7 Ab. In this study, mistletoe extract induced necrosis and apoptosis in the tumor mass was induced by HN22 cells, early and late apoptosis in vitro study. Mistletoe extract was likely to induce cell death in oral squamous cell carcinoma through apoptosis.

Expression of cytokine in CD69 cell and CD69 on lymphocyte from patients with systemic lupus erythematosus / 中国免疫学杂志

Objective:To investigate the expression of CD69+ on lymphocyte and cytokine in active lymphocyte (CD69+) from patients with systemic lupus erythematosus( SUE) . So try to understand the lymphocyte activation, expression of cytokine in active lymphocyte and the pathogenesis of SLE.Methods: Expression of CD69+ on lymphocyte and intracellular cytokine(IL-2,TNF?) in active lymphoycte(CD69+) were investigated in 26 SLE and 15 healthy volunteers by using flow cytometric analysis and immunofluorescence staining method. Results:Expression of CD69+ on lymphocytes from active and inactive SLE patients were significantly higher than that of healthy controls( P