Cation-osmotic haemolysis in stroke patients

Erythrocyte microrheology changes were measured by cation-osmotic haemolysis (COH) in healthy donors, patients with ischameic stroke, and patients who died within four days after a stroke. COH in patients with stroke was significantly decreased in comparison with that from healthy donors. In patients who died, COH was significantly decreased compared to patients who survived. The relationship between cationosmotic haemolysis and erythrocyte deformability is discussed.     

Cation-osmotic Haemolysis in Patients with Retinal Vein Occlusion

The purpose of this work was to use a new technique to assess erythrocyte deformability in patients with retinal vein occlusion. Erythrocyte microrheology changes were measured by cation-osmotic haemolysis (COH) in healthy donors and in patients with both central (n= 6) and branch (n= 16) retinal vein occlusion up to 12 months after the occlusion. The patient group consisted of five patients with ischaemic and 17 with oedematous vein occlusion. The control group consisted of the same number of age-, sex- and risk factor (hypertension)-matched donors. In patients with retinal vein occlusion, COH was significantly decreased compared to the control group. The decrease was most marked at low and high concentrations of incubating media where the differences reached high statistical significance (p<0.01–0.001). In our previous experiments we showed that COH and erythrocyte deformability (ED) are closely related and that COH reflects basic information about erythrocyte deformability. Thus, decreased COH in patients with retinal vein occlusion points to reduced ED in comparison with the control group. We, therefore, assume that impairment in ED together with other microrheological abnormalities contributes to the pathophysiology of changes in patients with retinal vein occlusion.     

Blood flow velocity,erythrocyte deformability and hepatic enzymes in heavy alcohol drinkers

Significantly decreased blood flow velocity in cerebral arteries, as well as decreased erythrocyte deformability, was found in heavy alcohol drinkers (Gdovinová 2002). The aim of this study was to determine if there is any correlation between the two. At the same time, the correlation between blood flow velocity and hepatic enzymes was also studied. The study group comprised 30 male patients, who were heavy alcohol drinkers, with a mean age of 46.4 years. Mean flow velocity (Vmean) was determined by a 2 MHz pulsed Doppler probe. Erythrocyte membrane biophysical properties were estimated by cation-osmotic haemolysis (COH). Hepatic enzymes [aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma-glutamyltransferase (GMT)] were measured in the Department of Biochemistry. The results were compared with the results from 20 healthy volunteers of the same age. Differences between the two groups were tested by Students t-test, and the relationship between blood flow velocity and COH, and of hepatic enzymes was analysed by regressive and correlative analysis. Results showed that Vmean was significantly decreased in heavy alcohol drinkers, in all cerebral arteries (the middle, anterior, and posterior cerebral arteries). No differences were noted between the left and right sides of the arteries tested. COH was similarly significantly decreased in heavy drinkers, in the medium of low (15.4 mmol/l NaCl) and high (123.2 mmol/l NaCl and above) ionic strength. Linear correlation between COH and Vmean was determined by correlative analysis, and a linear relationship was found. All hepatic enzymes (AST, ALT, GMT) were significantly increased in heavy drinkers. The relationship between Vmean and GMT was tested by the regressive analysis, and between these two parameters a linear relationship was found. It was concluded that changes in blood viscosity after the drinking of alcohol disturbs blood velocity and brain perfusion, which can be a major risk factor in strokes.     

Cerebral Blood Flow Velocity,Erythrocyte Deformability and Alcohol Intake

The aim of the study was to determine if there is a relationship between low blood flow velocity in the cerebral arteries and erythrocyte deformability in heavy alcohol drinkers. The study comprised 47 heavy alcohol drinkers (mean age 47 years). All of them drank daily more than 84 g of alcohol (84–400 g). Blood flow velocity (V _mean) in intracranial arteries was determined by transcranial Doppler. Erythrocyte membrane biophysical properties were estimated using the method of cation-osmotic haemolysis (COH). The present study revealed a significant decrease in V _mean in all examined arteries, with p= <0.01 in the middle (MCA) and posterior (PCA) cerebral arteries and p= <0.05 in the anterior cerebral artery (ACA) when compared with age-matched controls. Cation-osmotic haemolysis in the low ionic strength of the incubating medium (15.4 mmol/l NaCl) as well as in the high ionic strength (123.2–154.0 mmol/l NaCl) was significantly decreased (p<0.001–0.01). This means that changes in both parts of the erythrocyte membrane (actin–spectrin complex and membrane lipid bilayer) are the cause of decreased erythrocyte deformability. We conclude that one of the factors which can cause low blood flow velocity (a possible risk factor for stroke) is decreased cation-osmotic haemolysis of erythrocytes.     

急性脑卒中患者血清氧化应激指标测定及临床意义

目的:分析急性脑卒中患者氧化应激指标水平变化及其临床意义。方法:采用生物化学方法检测90例急性脑卒中患者(病例组)和50例健康体检者(对照组)血清超氧化物歧化酶(SOD)、丙二醛(MDA)水平和总抗氧化能力(T-AOC),并将病例组进一步分为脑出血组和脑梗死组,比较两组氧化应激指标的差异。结果:病例组血清SOD和T-AOC水平明显降低,MDA水平明显升高,差异均有统计学意义(P<0.05)。与脑梗死组相比,脑出血组血清SOD水平显著降低(P<0.05),MDA水平明显升高(P<0.05),T-AOC水平变化无明显差异(P>0.05)。结论:急性脑卒中患者抗氧化能力明显降低,氧化损伤作用增强。     

Some aspects of adrenocortical stress response following stroke

Stress response measured as the cortisol secretion rate as well as urinary 17 oxogenic steroid (17 OGS) and 17 oxosteroid (17 OS) excretion was investigated in a group of 28 patients with stroke (11 with hypertensive intracerebral hemorrhage and 17 with ischemic cerebrovascular insult). Significantly higher cortisol secretion values were found in the group of patients who died (p less than 0.01). Similarly, the first day urinary 17 OGS excretion pointed to the greater adrenocortical response in patients who died (p less than 0.05). Urinary 17 OS excretion was normal or below normal in the majority of patients and no difference was found between either male or female patients who survived or died. Our findings indicate that adrenocortical changes which maximize the production of cortisol are operative soon after the onset of stroke. The cortisol secretion rate appeared to be a good indicator of the severity of the stress caused by stroke and may be useful in predicting the prognosis of the illness.     

p56lck activity and expression in peripheral blood lymphocytes from patients with systemic lupus erythematosus.

In this study we analyzed the activity and the expression of p56lck protein tyrosine kinase in peripheral blood lymphocytes (PBLs) from systemic lupus erythematosus (SLE) patients and from healthy donors. The p56lck activity, determined by a non-radioactive Tyrosine Kinase Assay Kit, was significantly higher in active SLE PBLs and discriminated this group of patients from inactive SLE patients (p = 0.002) and healthy donors (p = 0.009). p56lck level decreased in SLE lymphocytes (especially for inactive SLE lymphocytes, p = 0.005) when compared to healthy donors. These differences were also reflected by the specific activity of p56lck that was clearly elevated in active SLE lymphocytes when compared to inactive SLE (p = 0.022) or healthy donors lymphocytes (p = 0.006). A positive correlation between the activity of p56lck and the tyrosine phosphorylation level in active SLE lymphocytes was found.     

Increased tumour necrosis factor alpha production by neutrophils in patients with hepatitis B.

AIMS--To investigate the role of serum and neutrophil tumour necrosis factor alpha (TNF alpha) in patients with viral hepatitis. METHODS--The activities of serum and neutrophil TNF alpha were measured using a bioassay of in vitro cytotoxicity against L929 cells in 57 patients with viral hepatitis and 20 healthy blood donors. RESULTS--Both serum and neutrophil TNF alpha in patients with chronic active hepatitis (CAH) and subacute fulminant hepatitis (SAFH) increased compared with those in normal controls (p < 0.01). No such differences were seen in patients with acute hepatitis. Serum and neutrophil TNF alpha were obviously reduced in patients with CAH and SAFH during convalescence compared with the active period (p < 0.05; p < 0.01). Furthermore, serum TNF alpha was significantly increased in patients with SAFH and complications compared with those without (p < 0.01), and in patients with SAFH who died compared with those who survived (p < 0.01). Neutrophil TNF alpha was significantly higher in patients with SAFH and secondary bacterial infections (p < 0.05). CONCLUSIONS--Production of serum and neutrophil TNF alpha is increased in patients with CAH and SAFH, suggesting that neutrophil TNF alpha causes liver injury in these patients.     

Ovarian response to standard gonadotrophin stimulation for IVF is decreased not only in older but also in younger women in couples with idiopathic and male subfertility

BACKGROUND: With the occasional reports of unexpectedly poor ovarian response to controlled ovarian hyperstimulation (COH) for IVF in young normally cyclic women in mind, we studied age-related ovarian response to COH in a group of women who underwent standard IVF. METHODS: Ovarian response to COH was defined as the number of follicles > or = 14 mm on the day of hCG administration. Ovarian response to COH was analysed by multiple regression analysis with woman's age and basal FSH concentration as explanatory variables in a prospective cohort of patients with idiopathic and mild male factor subfertility (n = 85), and additionally in a large retrospective cohort of women with unexplained, mild male and tubal subfertility (n = 1155), with age as explanatory variable. RESULTS: Ovarian response to COH was associated significantly with age (P < 0.001) and basal FSH concentration (P = 0.002). However, in women with idiopathic or mild male subfertility, in both cohorts the relationship took the form of an inverted U-shape with both older and--surprisingly--young women having a reduced ovarian response (P < 0.001). Maximum ovarian response was around the age of 28 years. In women with tubal infertility, there was only a linear decline of ovarian response with age. CONCLUSION: It is hypothesized that diminished ovarian response to COH in IVF is the very first sign of ovarian ageing in young women diagnosed with idiopathic and mild male subfertility.     

Circulating CD3+ CD4+ CD+ T lymphocytes in multiple sclerosis

Triple-antibody flow cytometry was used to search for distinctive populations of peripheral blood lymphocyte immunophenotypes in multiple sclerosis (MS). Using monoclonal antibodies to the cell surface markers CD3, CD4, and CD8, T cell subsets were quantified on a cohort of 31 MS patients (not treated with corticosteroids for at least 6 months), 30 healthy donors, and 14 patients with other autoimmune diseases (also corticosteroid treatment-free for at least 6 months). Untreated MS patients displayed a significantly greater population of CD3+CD4+CD8+ circulating T cells than healthy donors (P = 0.023). Patients with other autoimmune diseases displayed mean populations of CD3+CD4+CD8+ cells greater than normal donors and less than MS, but not significantly different from either. An additional 45 MS patients who had received corticosteroid therapy within the previous 6 months were phenotyped. Treatment of symptomatic MS with corticosteroids was associated with a smaller population of circulating CD3+CD4+CD8+ cells. Some MS patients have significantly greater numbers of peripheral blood T lymphocytes simultaneously expressing CD3, CD4, and CD8 surface markers than healthy donors and this population of cells may be reduced by corticosteroids treatment. This triple positive phenotype may be a manifestation of a systemic immune abnormality in MS.     

Altered immune function of monocytes in different stages of patients with acute on chronic liver failure

The aim of this study was to investigate the characteristics of the immune function of monocytes in different stages of the patients with acute on chronic liver failure (ACLF). Human leucocyte antigen (HLA)-DR and Toll-like receptor 4 (TLR-4) expression on monocytes in early and late stages of acute on chronic liver failure were detected by flow cytometry. The secretion function of monocytes was measured by cytometric bead array. Compared with healthy controls, the levels of HLA-DR expression on monocytes in patients with chronic hepatitis B, liver cirrhosis and acute on chronic liver failure were gradually decreased, especially in the late stage of acute on chronic liver failure (P < 0.001). TLR-4 expression on monocytes in patients with liver cirrhosis and acute on chronic liver failure were higher than the healthy controls. The concentrations of interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha and IL-12p70 in early-stage ACLF were significantly higher compared with healthy controls and lower in late-stage ACLF (P < 0.01, 0.05). However, a significantly lower amount of IL-10 was found on monocytes in early-stage ACLF than that of late-stage ACLF and healthy controls (P < 0.01). Monocyte HLA-DR expression in patients who died was significantly lower compared with patients who survived in the early and late stages of ACLF (P < 0.01). The dynamic detection of HLA-DR expression or cytokines secreted from monocytes could contribute to the estimation of the status of the immune function of patients with acute on chronic liver failure.     

Prothymosin αl effects in vitro on chemotaxis,cytotoxicity and oxidative response of neutrophils from melanoma,colorectal and breast tumor patients

Immunoregulatory effects of thymic peptides on functions of polymorphonuclear leukocytes (PMNs) are poorly investigated. We studied the effects of prothymosin α1 (Pro αl) on PMNs from patients with colorectal tumors, breast tumors and melanoma (total n = 37) in comparison with healthy donors (n = 18), with respect to chemotaxis, cytotoxicity against HCT-116 colon tumor cells, oxidative response (chemiluminescence reaction) as well as expression of surface marker molecules. We found that Pro α1 was equally effective in stimulating the chemotactic activity of PMNs from tumor patients and healthy donors (43% increase). PMNs from tumor patients, especially with breast tumor, showed a significant enhancement of cytotoxicity against the tumor target cells in comparison with healthy donors. With respect to the PMNs cytotoxicity, only about 50% of the colorectal tumor patients and healthy donors responded to Pro α1 and FMLP. As to the oxidative response of PMNs, elevated levels were found only among colorectal tumor patients. Pro α1 significantly increased the oxidative response in breast and colorectal tumor patients by 55% and 25%, respectively. Pro α1 decreased the expression of CD16 on PMNs of healthy donors, but not that of CD11a, CD1lb, CD1lc, CD13, CD14, CD15 and CD32. Therefore, we suggest, that Pro α1 may improve some PMN functions of tumor patients, associated with the proposed role in host-tumor interaction.     

Immune responses to bile-tolerant helicobacter species in patients with chronic liver diseases,a randomized population group,and healthy blood donors

Bile-tolerant Helicobacter species such as Helicobacter pullorum, Helicobacter bilis, and Helicobacter hepaticus are associated with hepatic disorders in animals and may be involved in the pathogenesis of chronic liver diseases (CLD) in humans. Antibody responses to cell surface proteins of H. pullorum, H. bilis, and H. hepaticus in serum samples from patients with CLD, a randomized population group, and healthy blood donors were evaluated by using enzyme linked immunosorbent assay (ELISA). The results were compared with the antibody responses to Helicobacter pylori. For analysis of a possible cross-reactivity between bile-tolerant Helicobacter species and H. pylori, sera from a subpopulation of each group were absorbed with a whole-cell extract of H. pylori and retested by ELISA. Results before absorption showed that the mean value of the ELISA units for H. pullorum was significantly higher in patients with CLD than in healthy blood donors (P = 0.01). Antibody reactivity to cell surface protein of H. hepaticus was also significantly higher in the CLD patients than in the healthy blood donors and the population group (P = 0.005 and P = 0.002, respectively). Following the absorption, antibody responses to H. pullorum decreased significantly in all three groups (P = 0.0001 for CLD patients, P = 0.0005 for the population group, and P < 0.0001 for the blood donors), indicating that cross-reactivity between H. pylori and other Helicobacter spp. occurs. The antibody responses to H. hepaticus and H. bilis in CLD patients remained high following absorption experiments compared to ELISA results before absorption. The significance of this finding requires further investigations.     

Serologic survey for exposure following fatal Balamuthia mandrillaris infection

Granulomatous amebic encephalitis (GAE) from Balamuthia mandrillaris, a free-living ameba, has a case fatality rate exceeding 90 % among recognized cases in the USA. In August 2010, a GAE cluster occurred following transplantation of infected organs from a previously healthy landscaper in Tucson, AZ, USA, who died from a suspected stroke. As B. mandrillaris is thought to be transmitted through soil, a serologic survey of landscapers and a comparison group of blood donors in southern Arizona was performed. Three (3.6 %) of 83 serum samples from landscapers and 11 (2.5 %) of 441 serum samples from blood donors were seropositive (p?=?0.47). On multivariable analysis, county of residence was associated with seropositivity, whereas age, sex, and ethnicity were not. Exposure to B. mandrillaris, previously unexamined in North America, appears to be far more common than GAE in Southern Arizona. Risk factors for disease progression and the ameba’s geographic range should be examined.     

乙型病毒性肝炎患者血清sICAM-1水平及意义

为探讨乙型肝炎患者血清ｓＩＣＡＭ－１水平及其与肝损害的关系，用酶联免疫法检测７例正常人和７４例ＨＢＶ感染血清ｓＩＣＡＭ－１。结果：重型肝炎（重肝）早、中、晚期，及轻、中、重度慢性乙型肝炎（ＣＨＢ）患者血清ｓＩＣＡＭ－１，均显著高于正常人和无症状ＨＢｓＡｇ携带者（ＡｓＣ）；ＣＨＢ患者肝功能越差、其血清ｓＩＣＡＭ－１水平越高，而重肝患者肝功能越差、其血清ｓＩＣＡＭ－１水平也越低。表明：从ＡｓＣ至重肝早期，血清ｓＩＣＡＭ－１水平随着患者肝损害的加重而升高，能较好反映肝组织炎症坏死程度，重肝患者血清ｓＩＣＡＭ－１水平随着肝衰竭的加重而降低。     

A new CD21low B cell population in the peripheral blood of patients with SLE

A hallmark of systemic lupus erythematosus (SLE) is the production of autoantibodies. Recent reports suggest an abnormal peripheral blood B cell homeostasis in SLE patients without being conclusive. We analyzed by four color flow-cytometry peripheral blood B cell subpopulations of SLE patients, healthy donors, and patients with other systemic autoimmune diseases. IgM memory but not switched memory B cells of SLE patients were significantly decreased compared to healthy donors, whereas transitional B cells, characterized by CD19+IgMhiIgD+CD24hiCD38hi, were significantly expanded in SLE patients but also found in other autoimmune disorders. The population of plasmablasts (CD19loCD21loCD27++CD38++) was increased in active disease. Most interestingly, B cells in autoimmune disorders contain a so far uncharacterized subpopulation with an activated phenotype (CD19hiCD21loCD38loCD86int). None of the identified subpopulations was associated with current or previous therapy and therefore may represent different aspects of the disturbed B cell homeostasis in patients with SLE.     

CD4+CD25+Treg在急性淋巴细胞白血病患者外周血中的表达及其体外实验研究

目的初步探讨CD4 CD25 调节性T细胞(CD4 CD25 regulatory T cells,CD4 CD25 Treg)在急性淋巴细胞白血病(acute lymphocytic leukemia,ALL)患者化疗前及化疗缓解后外周血中的表达水平,并研究患者血清能否诱导外周血CD4 CD25-T细胞转化为CD4 CD25 Treg。方法①采用流式细胞术分别检测ALL初诊组、化疗完全缓解或部分缓解组及正常对照组外周血中CD4 CD25 T细胞所占比例,然后通过荧光定量RT-PCR检测各组外周血中转录因子Foxp3mRNA的表达水平,并逐层分析比较。②采集正常人外周血单个核细胞后,对照组用正常人血清,实验组用患者血清并分别设浓度梯度进行培养,72h后采用流式细胞术、荧光定量RT-PCR分别检测CD4 CD25 T细胞和Foxp3mRNA表达。结果ALL化疗缓解组CD4 CD25 T细胞及Foxp3mRNA表达水平均明显高于ALL初诊组和正常对照组(P<0.05),后两者之间CD4 CD25 T细胞水平无统计学差异(P>0.05),但ALL初诊组Foxp3mRNA含量较正常对照组明显升高(P<0.01),差异具有统计学意义;并且血清培养对照组CD4 CD25 T细胞水平及Foxp3mRNA含量均明显低于实验组(P<0.05),且其表达并不随血清浓度的增加而升高。结论CD4 CD25 Foxp3 Treg在ALL初诊组及化疗缓解组患者外周血中比例明显升高,且初步表明患者血清中的可溶性物质可诱导外周血CD4 CD25 T细胞转化为CD4 CD25 Treg,提示CD4 CD25 Treg可能是ALL免疫抑制的一个重要原因。     

淋巴细胞化学发光临床意义的初探

用ConA诱导人外周血淋巴细胞化学发光(Ly-CL)方法,测定112例正常人和180例不同性质疾病患者(包括肿瘤78例、银屑病39例,带状疱疹18例、病毒性心肌炎18例、湿疹17例和荨麻疹10例)Ly-CL变化。结果表明:①正常人Ly-CL变化与性别无关,而与年龄因素有关,中老年者Ly-CL低于年青者;②与相近年龄的正常人相比,肿瘤、带状疱疹、银屑病和病毒性心肌炎患者Ly-CL降低,而荨麻疹Ly-CL增高。本文结果提示,人Ly-CL变化与淋巴细胞活性或功能之间存在一定联系,不同性质疾病患者淋巴细胞产生氧自由基能力有差异。     

Secretion of interleukin-2 (IL-2) and interferon (IFN gamma)in whole blood cell culture stimulated with mitogens in patients with lung neoplasms

The aim of this study was to compare the ability of blood lymphocytes from lung cancer patients to secrete interleukin-2 (IL-2) and interferon gamma (IFNg) upon stimulation with mitogens with that of healthy donors. 42 patients with small cell lung cancer (SCLC), 30 patients with non small cell lung cancer (NSCLC) and 30 healthy donors were studied. The test was done in lung cancer patients before treatment. IL-2 and IFNg levels were measured with Elisa ready kits (Genzyme) in the supernatants of whole blood culture after stimulation with Pokeweed (PWM) and Phytohemagglutinin (PHA) mitogens. The results of the cytokine levels after stimulation were not normally distributed and thus were transformed to logarthms for statistical evaluation. The t-test for transformed results were used to asses the difference between groups. The median level of IFNg in the supernatant of whole blood cultures was significantly lower in lung cancer patients than in healthy blood donors both after PWM as well as after PHA stimulation. When patients with NSCLC and SCLC were regarded separately the lower level of IFNg in comparison with healthy donors was found in the supernatant of the blood cultures only after stimulation with PWM. The median level of IL-2 in the supernatant of whole blood culture in lung cancer patients was lower than in healthy blood donors only after PWM stimulation. The same was true for SCLC patients. In NSCLC IL-2 levels were significantly lower after stimulation with PWM as well after PHA stimulation. In conclusion: secretion of IL-2 and IFNg in whole blood culture after mitogen stimulation in lung cancer patients is significantly lower than in healthy donors. No significant differences between SCLC and NSCLC were found.