Expression of L-Selectin in Peripheral Blood Leukocytes of Crossbred Cows Affected with Subclinical Mastitis

L-Selectin (SELL) linked with innate immune mechanism involved in neutrophil migration through endothelium to the site of inflammation. Early recruitment of neutrophils at the site of infection is essential to counteract infection at the earliest in conditions like sub-clinical mastitis (SCM). The present study was framed to evaluate the expression pattern of SELL in naturally occurring SCM in crossbred animals using real time PCR technique. Analysis of data on total leukocyte count indicated leucocytosis condition in SCM affected crossbred cows. Relative expression of SELL on peripheral blood leukocytes revealed a significant 3.16 folds down regulation in SCM affected cows when compared with healthy crossbred cows (P<0.05). The possible reason for leukocytosis in SCM animals might be due to down regulation of SELL on leukocytes reducing their ability to transmigrate through blood vessel to infection site. The result of present study revealed a definite role of SELL in SCM which could be explored for therapeutic aspects in near future

Relationships between blood leukocyte mitochondrial DNA copy number and inflammatory cytokines in knee osteoarthritis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Osteoarthritis (OA) is a degenerative articular disorder manifested by cartilage destruction, subchondral sclerosis, osteophytes, and synovitis, resulting in chronic joint pain and physical disability in the elderly. The purpose of this study was to investigate mitochondrial DNA copy number (mtDNACN) and inflammatory cytokines in primary knee OA patients and healthy volunteers. A total of 204 knee OA patients and 169 age-matched healthy volunteers were recruited. Their relative blood leukocyte mtDNACN was assessed by quantitative real-time polymerase chain reaction (qRT-PCR), and ten inflammatory cytokines in their plasma were detected by multiplex immunoassay. Blood leukocyte mtDNACN in the OA group was significantly lower than that in the control group. Leukocyte mtDNACN in the control group was negatively correlated with their age (r=−0.380, P<0.0001), whereas mtDNACN in the OA group was positively correlated with their age (r=0.198, P<0.001). Plasma interleukin-4 (IL-4) and IL-6 were significantly higher in the knee OA group than in the control group. The plasma IL-6 level was positively correlated with blood leukocyte mtDNACN in the OA group (r=0.547, P=0.0014). IL-5 showed as a major factor (coefficient 0.69) in the second dimension of principle components analysis (PCA)-transformed data and was significantly higher in the OA group (P<0.001) as well as negatively correlated with mtDNACN (r=−0.577, P<0.001). These findings suggest that elevation of plasma IL-4 and IL-6 and a relative reduction in mtDNACN might be effective biomarkers for knee OA. IL-5 is a plausible factor responsible for decreasing blood leukocyte mtDNACN in knee OA patients.

Relationships between blood leukocyte mitochondrial DNA copy number and inflammatory cytokines in knee osteoarthritis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Osteoarthritis (OA) is a degenerative articular disorder manifested by cartilage destruction, subchondral sclerosis, osteophytes, and synovitis, resulting in chronic joint pain and physical disability in the elderly. The purpose of this study was to investigate mitochondrial DNA copy number (mtDNACN) and inflammatory cytokines in primary knee OA patients and healthy volunteers. A total of 204 knee OA patients and 169 age-matched healthy volunteers were recruited. Their relative blood leukocyte mtDNACN was assessed by quantitative real-time polymerase chain reaction (qRT-PCR), and ten inflammatory cytokines in their plasma were detected by multiplex immunoassay. Blood leukocyte mtDNACN in the OA group was significantly lower than that in the control group. Leukocyte mtDNACN in the control group was negatively correlated with their age (r=-0.380, P<0.0001), whereas mtDNACN in the OA group was positively correlated with their age (r=0.198, P<0.001). Plasma interleukin-4 (IL-4) and IL-6 were significantly higher in the knee OA group than in the control group. The plasma IL-6 level was positively correlated with blood leukocyte mtDNACN in the OA group (r=0.547, P=0.0014). IL-5 showed as a major factor (coefficient 0.69) in the second dimension of principle components analysis (PCA)-transformed data and was significantly higher in the OA group (P<0.001) as well as negatively correlated with mtDNACN (r=-0.577, P<0.001). These findings suggest that elevation of plasma IL-4 and IL-6 and a relative reduction in mtDNACN might be effective biomarkers for knee OA. IL-5 is a plausible factor responsible for decreasing blood leukocyte mtDNACN in knee OA patients.

Changes of Neutrophil Morphological Parameters in Peripheral Blood of Patients with Solid Tumor Treated with Granulocyte Colony Stimulating Factor / 现代检验医学杂志

Objective To study neutral particle particle(NEUT-X)change in the solid tumor patients with chemotherapy by granulocyte colony-stimulating factor(G-CSF).Methods Chose that 52 cases of cancer chemotherapy with G-CSF(study group),32 cases of cancer chemotherapy patients without G-CSF(study control group)and 50 cases of healthy(healthy control group).The automatic hematology analyzer Sysmex XE-2100 were been examined the peripheral blood routine and collected the data which wes the morphological parameters of peripheral white blood cell.The changes of neutrophil N-X pa-rameters during chemotherapy were analyzed,and the clinical infection fever rates of three groups were collected to reveal the relationship between leukocyte morphological parameters and body resistance.Results In the study group,study control group and healthy control group,the NEUT-X was 1 324(890.2,1 358.0),1 440(1 397.3,1 466.3)and 1 329(1 295.1, 1 359.4),and the difference was statistically significant between the three groups(F=10.778,P=0.002).In study group, the count of WBC before and after G-CSF was 0.99(0.22,1.75)×109/L and 7.53(1.00,14.05)×109/L respitively and there was the significant difference(Z=-2.395,P=0.005).In study group patients the NEUT-X was 1 382(1 323.6,1 440.4)and 1 324(890.2,1 358.0)respectively and there was a significant difference(Z=-2.832,P=0.004).Between the study group and the study control group,there were 23/52 cases and 4/32 cases infection in patients with fever case(Z=9.14,P=0.002).Conclusion By G-CSF the leukocyte counts increased in patients with chemotherapy,and reduced neu-trophil NEUT-X parameters,and the infection rate was higher than the non G-CSF patients.The neutrophil granularity will be useful for evaluating of patients with chemotherapy for solid tumor immunity.

Association between optical coherence tomography characteristics and peripheral blood leukocyte count in patients with acute myocardial infarction / 中国介入心脏病学杂志

Objective To explore the relationship between optical coherence tomography (OCT) characteristics and peripheral blood leukocyte count in patients with acute myocardial infarction(AMI). Methods A total of 33 patients with AMI hospitalized in Fuwai Hospital for primary percutaneous coronary intervention were consecutively enrolled,and underwent intracoronary OCT procedures after manual aspiration of coronary thrombus. Demographic data, risk factors, procedural and OCT data, past medical history and perioperative laboratory findings were collected in all patients. Results The lymphocyte count,monocyte count and basophil count were significantly higher in the patients with fibrous cap thickness ≥ 65 μm than in those patients with fibrous cap thickness ≤ 65 μm. Patients presenting with cholesterol crystallization had lower eosinophil count than those patients with crystallization-free [(0.04±0.06)×109/L vs.(0.10±0.09) ×109/L,P =0.028]. In addition,the former had significantly higher to neutrophil lymphocyte ratio than the latter [(8.35±6.13)vs.(4.97±2.01), P =0.020]. Higher monocyte count was found in the patients with calcified plaque (P <0.05). Platelet to lymphocyte ratio was significantly increased in the patients with macrophage infiltration [(165.72±85.93)vs.(113.47±19.13),P <0.05]. The leukocyte count,neutrophil count and monocyte count showed the treat of elevation as the number of OCT characteristics increased,but only the elevation of monocyte count had statistical significant level (P = 0.014). Conclusions Peripheral blood leukocyte count seems to be associated with OCT characteristics of plaque rupture in patients with acute myocardial infarction,suggesting the potential role of inflammation in plaque rupture.

Hipometilação do LINE-1 está Associada ao Risco de Doença Cardíaca Coronariana na População Chinesa / LINE-1 Hypomethylation is Associated with the Risk of Coronary Heart Disease in Chinese Population

Fundamentos: O nível de metilação global do ADN de leucócitos no sangue tem sido associado ao risco de doença arterial coronariana (DAC), com resultados inconsistentes em diferentes populações. Faltam dados semelhantes da população chinesa, onde diferentes fatores genéticos, de estilo de vida e ambientais podem afetar a metilação do ADN e sua relação com o risco de DCC. Objetivos: Analisar se a metilação global está associada ao risco de doença coronariana na população chinesa. Métodos: Foram incluídos um total de 334 casos de DCC e 788 controles saudáveis. A metilação global do ADN de leucócitos de sangue foi estimada por meio da análise das repetições do LINE-1 usando pirosequenciamento de bissulfito. Resultados: Em uma análise inicial restrita aos controles o nível do LINE-1 diminui significativamente com a idade avançada, colesterol total elevado, e diagnóstico de diabetes. Na análise de caso-controle, a redução da metilação do LINE-1 foi associada ao aumento do risco de DCC, tendo a análise por quartil revelado uma odds ratio (IC 95%) de 0,9 (0,6-1,4), 1,9 (1,3-2,9) e 2,3 (1,6 3.5) para o terceiro, segundo e primeiro (o mais baixo) quartil (P da tendência < 0,001), respectivamente, em comparação com o quarto (o mais alto) quartil. A metilação inferior (< mediana) do LINE-1 esteve associada a 2,2 vezes (IC 95% = 1,7-3,0) o aumento de risco de doença coronariana. As estimativas de risco de DCC menores relacionadas com o LINE-1 tenderam a ser mais fortes entre os indivíduos com maior tercil de homocisteína (P interação = 0,042) e naqueles com diagnóstico de hipertensão arterial (P interação = 0,012). Conclusão: A hipometilação do LINE-1 está ...

Background: Global methylation level in blood leukocyte DNA has been associated with the risk of coronary heart disease (CHD), with inconsistent results in various populations. Similar data are lacking in Chinese population where different genetic, lifestyle and environmental factors may affect DNA methylation and its risk relationship with CHD. Objectives: To examine whether global methylation is associated with the risk of CHD in Chinese population. Methods: A total of 334 cases with CHD and 788 healthy controls were included. Global methylation in blood leukocyte DNA was estimated by analyzing LINE-1 repeats using bisulfite pyrosequencing. Results: In an initial analysis restricted to control subjects, LINE-1 level reduced significantly with aging, elevated total cholesterol, and diagnosis of diabetes. In the case-control analysis, reduced LINE-1 methylation was associated with increased risk of CHD; analysis by quartile revealed odds ratios (95%CI) of 0.9 (0.6-1.4), 1.9 (1.3-2.9) and 2.3 (1.6-3.5) for the third, second and first (lowest) quartile (Ptrend < 0.001), respectively, compared to the fourth (highest) quartile. Lower (<median) LINE-1 methylation was associated with a 2.2-fold (95%CI = 1.7-3.0) increased risk of CHD. The lower LINE-1-related CHD risk estimates tended to be stronger among subjects with the highest tertile of homocysteine (Pinteraction = 0.042) and those with diagnosis of hypertension (Pinteraction = 0.012). Conclusion: LINE-1 hypomethylation is associated with the risk of CHD in Chinese population. Potential CHD risk factors such as older age, elevated total cholesterol, and diagnosis of diabetes may have impact on global DNA methylation, whereby exerting their effect on CHD risk. .

Study of high throughput screening for inhibitors of inflammatory cytokine / 中国药理学通报

AIM The aim of our study was to establish drug screening models which can evaluate samples' effects on cytokines related with inflammtion, capable of fast and efficient screening of anti-inflammatory lead compounds on the release of inflammtory cytokines. METHODS Heparinized human blood leukocytes was evaluated as a model to study the effects of various classes of anti-inflammatory lead compounds on cytokine release/biosynthesis from leukocytes. Human blood leukocytes was stimulated with LPS (final concentration 0.5～50 mg?L -1), with or without test drugs (diclofenac, a cytooxygenase inhibitor, nordihydroguaiaretic acid NDGA, a 5-lipoxygenase inhibitor) for 1～4 h to induce cytokine release. RESULTS Human blood leukocytes stimulated with LPS could product IL-1, IL-8 and TNF-? in a dose-dependent manner. Human blood leukocytes was stimulated with LPS(5 mg?L -1) for 4h to induce cytokine release. TNF-?, IL-1 and IL-8 time-course profiles were determined in culture media, using bioassays and ELISA. LPS-mediated release of IL-1 and TNF-? was significantly suppressed by NDGA and Diclofenac. In LPS stimulated blood, NDGA and Diclofenac inhibited the release of TNF-?(IC 50 of 149 ?mol?L -1 and 23.88 ?mol?L -1) or IL-1 (IC 50 of 222.57 ?mol?L -1 and 126 ?mol?L -1). CONCLUSION This human blood leukocytes screening system in vitro has the potential to screen new cytokine release inhibitors and sites of action of new anti-inflammatory lead compounds, and increases the screening efficiency.