多发性骨髓瘤患者骨髓细胞IL-6分泌机制的研究

目的：探讨多发性骨髓瘤（ＭＭ）患者骨髓细胞中白细胞介素６（ＩＬ－６）分泌机制。方法：应用双标记免疫荧光技术检测了１６例ＭＭ患者和１９例对照者新鲜分离的骨髓单个核细胞中ＩＬ－６表达情况。结果：ＭＭ患者骨髓单个核细胞ＩＬ－６表达阳性率为１５．４％±６．５％，骨髓瘤细胞ＩＬ－６表达阳性率达７．９％±２．９％，基质细胞ＩＬ－６表达阳性率达４．５％±１．２％；对照组骨髓单个核细胞ＩＬ－６表达阳性率为４．６％±２．３％，基质细胞ＩＬ－６表达阳性率为２．６％±１．１％，浆细胞ＩＬ－６表达阳性率为０．７％±０．１％。ＭＭ患者均较对照组显著升高（Ｐ均＜０．００１）。另外ＭＭ组骨髓单个核细胞中基质细胞所占比率为６．９％±１．４％，也较对照组的４．７％±１．８％明显升高（Ｐ＜０．００１）。结论：ＭＭ患者升高的ＩＬ－６既可由瘤细胞分泌，又可由基质细胞分泌，自分泌和旁分泌机制可能都参与了ＭＭ的发病。     

Update on value of the anion gap in clinical diagnosis and laboratory evaluation

Anion gap (AG) is a calculated value commonly used in clinical practice. It approximates the difference between the concentration of unmeasured anions (UA) and unmeasured cations (UC) in serum. At present, the reference range of anion gap has been lowered from 8-16 to 3-11 mmol/l because of the changes in technique for measuring electrolyte. However, clinicians and textbooks still refer and use the old reference value of 8-16 mmol/l. This may lead to misinterpretation of the value of anion gap. Our study updated the value of anion gap in clinical diagnosis and laboratory evaluation. Criteria for using anion gap were also suggested. We analyzed serum electrolyte using the Beckman Synchron CX5. The anion gap was calculated from the formula: [Na(+)-(Cl(-)+HCO(3)(-))]. We estimated the reference range using the non-parametric percentile estimation method. The reference range of anion gap obtained from 124 healthy volunteers was 5-12 mmol/l, which was low and confirmed the reports from other studies (3-11 mmol/l) using ion-selective electrode. From the retrospective study on the 6868 sets of serum electrolyte among hospitalized patients, we found the incidences of normal, increased, and decreased anion gaps were 59.5%, 37.6%, and 2.9%, respectively. The mean and central 90% range of increased anion gap were 16 and 13-20 mmol/l, which was lower than those reported in previous study (25 and 19-28 mmol/l). Anion gap exceeding 24 mmol/l was rare. The mean and central 90% range of decreased anion gap were 3 and 2-4 mmol/l, which were lower than those reported in previous study (6 and 3-8 mmol/l). The value of less than 2 mmol/l was rare. The most common causes of increased anion gap (hypertensive disease, chronic renal failure, malignant neoplasms, diabetes mellitus and heart diseases) and decreased anion gap (liver cirrhosis and nephrotic syndrome) in this study were similar to those in previous studies. We found two cases of IgG multiple myeloma with anion gap of 2 mmol/l. In conclusion, clinicians and laboratorians can use the anion gap as clue in quality control. They can check the incidences of increased and decreased anion gap. If one finds high incidence of increased anion gap (>24 mmol/l) or decreased anion gap (<2 mmol/l), one should check the quality control of electrolyte and whether the patients were hypoalbuminemia or hyperglobulinemia. An anion gap exceeding 24 mmol/l will suggest the presence of metabolic acidosis. It is very rare to find anion gap with the negative sign.     

IgD型多发性骨髓瘤患者实验室检查结果分析

目的 IgD型多发性骨髓瘤患者临床上较为少见,通过实验室检查,以求发现其特点,避免漏诊。方法分别收集20例本实验室检测到的IgD型I、gG型I、gA型I、gM型多发性骨髓瘤患者的实验室检查结果,分析IgD型与其它较为常见类型的多发性骨髓瘤患者实验室检查结果的异同。结果 IgD型多发性骨髓瘤患者的免疫球蛋白定量结果显示,IgGI、gA和IgM含量均减低者有18例占90%,其余两例3种免疫球蛋白含量处于正常下限。轻链定量发现,20例IgD型多发性骨髓瘤患者有19例为IgD型λ轻链型,占95%,而IgD型κ轻链型仅1例,且20例中有19例是轻链比值异常的。IgD型多发性骨髓瘤患者的血清蛋白电泳阳性率与其余类型无显著差异（P〉0.05）;免疫固定电泳显示IgD泳道有特异性的抗原抗体沉淀带。结论如实验室检查发现患者免疫球蛋白IgGI、gA和IgM含量均减低、轻链比值异常、血清蛋白电泳出现阳性条带,要高度怀疑其为IgD型多发性骨髓瘤,免疫固定电泳时务必加IgD泳道,以避免IgD型多发性骨髓瘤的漏诊。     

Differential immunoadsorption coupled with rate nephelometry for estimation of DNA-binding immunoglobulins

We describe a technique for estimating the mass of anti-DNA antibodies by immunonephelometry of serum immunoglobulins (IgG, IgA, IgM) before and after adsorption onto DNA bound to agarose-polylysine columns. Sixteen patients with systemic lupus erythematosus and 16 age- and sex-matched controls were studied. Precision was determined for high-value (in 10 patients) and low-value (in nine controls) ranges for each of the immunoglobulins. Within-run CVs ranged from 3.0% (IgG, controls) to 11.8% (IgA, patients); between-run CVs ranged from 15.5% (IgG, patients) to 25.2% (IgM, patients). We found anti-DNA antibody concentrations (mean +/- SD) in systemic lupus erythematosus of 1.981 +/- 1.015 g/L for IgG (controls: 0.243 +/- 0.231, p less than 0.001), 0.257 +/- 0.215 g/L for IgA (controls: 0.038 +/- 0.035, p less than 0.001), and 0.282 +/- 0.234 g/L for IgM (controls: 0.191 +/- 0.165, p greater than 0.05). Sensitivity and linearity are such that fivefold dilutions of patients' serum with either a buffered albumin solution or control serum yielded values close to the expected values for IgG. Similarly diluted sera gave inordinately high values in the radiometric binding assay. Neither parametric (linear regression) nor nonparametric correlation methods (Spearman's rank and Kendall's tau) show a significant correlation between patients' data obtained by the present technique and that by a radiometric binding assay (p greater than 0.05), although combined data from patients and controls demonstrate a significant nonparametric correlation (p less than 0.005 for Spearman's and p less than 0.02 for Kendall's).     

Decreased anion gap in polyclonal hypergammaglobulinemia

OBJECTIVES: The anion gap has proved a valuable tool in the diagnosis of various forms of acid-base disorders, although the importance of slight rises in the anion gap remains unclear. The concept of the anion gap is often misunderstood and misapplied. The relationship between gammaglobulins and the serum anion gap has not received much attention except for reports of a narrowing of the gap associated with certain monoclonal immunoglobulin G gammopathies. We present patients with polyclonal gammopathy, the magnitude of which correlated strongly and negatively with the anion gap. DESIGN: The anion gap can be readily calculated from routine laboratory data, and anion gap was calculated as ([Na] +[K])- ([Cl] + [HCO3]). SUBJECTS: Serum anion gaps were determined in 206 patients with polyclonal hypergammaglobulinemia and 63 healthy subjects. MEASUREMENTS: Serum sodium and potassium ions concentration were determined by flame photometry. Serum bicarbonate level was measured as total carbon dioxide content. Serum chloride level was determined by chlorimetric titration with silver ions. RESULTS: All patients with polyclonal hypergammaglobulinemia had a statistically significant reduction in their mean serum anion gaps (6.4 +/- 1.2 mmol/L) when compared with normal control volunteers (15.3 +/- 2.4 mmol/L), p < 0.05. CONCLUSIONS: Our results show a negative correlation between serum anion gap and gammaglobulins concentration.     

Lower anion gap increases sensitivity in predicting elevated lactate.

OBJECTIVE: The normal reference range for the anion gap (AG) has recently been questioned by several authors. Lowering the upper limit of normal of the AG has been found to be more sensitive in predicting elevated lactate in critically ill adults. The objectives of this study are i) to define a new upper limit of normal of the AG in a study population of healthy adult volunteers, ii) to determine the sensitivity, specificity, the positive predictive value and the negative predictive value of the new upper limit for AG in detecting elevated lactate in critically ill children and to compare these results to the old upper limit of normal of AG (16 mmol/l), iii) to construct a receiver-operating-characteristic (ROC) curve for anion gap as a predictor of elevated lactate, iv) to determine the relationship between anion gap and serum lactate levels in critically ill patients. DESIGN: A prospective, cohort study. SETTING: Paediatric Intensive Care Unit of a University Hospital. SUBJECTS: Part I: Convenience sample of healthy adult volunteers to provide a reference range for anion gap calculation. Part II: Consecutive children admitted to the Paediatric Intensive Care Unit who had lactate levels measured for clinical reasons. MEASUREMENTS: Part I: Electrolytes and blood gases were measured from blood samples drawn from 25 adult volunteers. The reference range for AG was calculated using the equation, AG = Na - (Cl + HCO3). The upper limit of normal was calculated as mean + 2 SD. Part II: Eligible ICU patients were included in this study if they had lactate, electrolytes and blood gases obtained simultaneously. The AG was calculated as above. The new upper limit of normal AG was compared to an AG of 16 for diagnosing an elevated plasma lactate. RESULTS: The mean anion gap in the normal population was 9.4 +/- 1 mmol/l with 11 mmol/l being used as the new upper limit of normal. Thirty-six ICU patients had 189 arterial blood samples from which lactate, electrolytes and blood gas were measured simultaneously. The sensitivity, specificity, positive predictive value and negative predictive value of using an AG of 11 mmol/l as the upper limit of normal were 86%, 40%, 65% and 69% respectively, compared to 49%, 84%, 80% and 55% respectively using the upper limit of normal of AG of 16 mmol/l. The ROC curve supported lowering the upper limit of normal for the anion gap to predict an elevated lactate. There was a linear relationship between anion gap and serum lactate levels. CONCLUSIONS: An AG of 11 mmol/l as the upper limit of normal has a higher sensitivity and higher negative predictive value but lower specificity and lower positive predictive value for detecting elevated lactate in critically ill children.     

免疫固定电泳在多发性骨髓瘤辅助诊断中的应用价值

目的 探讨免疫固定电泳免疫一削减法在多发性骨髓瘤患者辅助诊断中的应用.方法 60例骨髓涂片确诊为多发性骨髓瘤患者,分别对其血清进行免疫固定电泳和特种免疫球蛋白定量测定,并对两组结果进行比较.结果 在60例MM患者中,免疫固定电泳检测结果显示IgG型占56.7％（34/60）,IgA型占16.7％（10/60）,IgM型占18.3％（11/60）,单纯轻链型占5％（3/60）.特种免疫球蛋白定量结果显示,在各型MM患者血清中相应免疫球蛋白升高的比例分别为：IgG为21/34,IgA为8/10,IgM为11/11.免疫固定电泳对IgG型和IgA型MM患者的检出率明显高于特种免疫球蛋白定量分析,差异具有统计学意义（P＜0.05）.结论 在多发性骨髓瘤的实验室诊断中,免疫固定电泳对多发性骨髓瘤的检出率优于特种免疫球蛋白定量检测.     

Preanalytical handling of samples for measurement of plasma lactate in HIV patients

Lactic acidosis is a feared side effect of nucleoside analog treatment, one of the cornerstones in the management of HIV infection. Precise and reliable lactate measurements are prerequisites for the diagnosis of hyperlactatemia. The effects of venous stasis, time to measurement and storage temperature on p-lactate levels, p-glucose levels, anion gap and pH were investigated. Ten HIV patients (n=8 on highly active antiretroviral therapy) and 4 healthy control subjects were studied. Blood was drawn without stasis at time 0 and with stasis for 2 and 8 min into heparin-preserved test tubes. The tubes were placed at a room temperature (25 degrees C) and on crushed ice and consecutively monitored for up to 360 min. The mean increases in p-lactate in blood kept in test tubes at 25 degrees C, measured from 0 to 60 min and from 240 to 360 min, were increased in HIV patients compared with controls (0.78 mmol/Lh +/- 0.02 vs. 0.63 mmol/Lh +/- 0.05, (p=0.009) and 0.65 mmol/Lh +/- 0.03 vs. 0.53 mmol/Lh +/- 0.05, (p=0.042)). It was found that placing the tubes on crushed ice rather than keeping them at 25 degrees C controlled glycolysis and lactate production measured over a 6-h period (0.033 mmol/Lh +/- 0.006 vs. 0.32 mmol/Lh +/- 0.01, (p<0.0001) and 0.064 mmol/Lh +/- 0.008 vs. 0.64 mmol/Lh +/- 0.02, (p<0.0001)). The total increases in lactate levels in the test tubes placed on crushed ice for 4 h and in those kept at 25 degrees C for 15 min were comparable (0.28 +/- 0.03 mmol/L vs. 0.20 +/- 0.03). Compared with storage at 25 degrees C, keeping the test tubes on crushed ice also preserved pH and anion gap over a 6-h measurement period (pH: 0.026 +/- 0.004 vs. 0.12 +/- 0.01 and anion gap: -0.8 +/- 0.4 mmol/L vs. 4.1 +/- 0.4). Two minutes of venous stasis had no influence on p-lactate levels (0.02 +/- 0.04 mmol/L, p=0.70), whereas 8 min of stasis increased p-lactate levels by 0.11 +/- 0.04 mmol/L, p=0.009. It is concluded that major errors in measurements of p-lactate, anion gap and pH can be prevented by placing test tubes on crushed ice for up to 4 h until measurement.     

血清κ/λ轻链比值及蛋白电泳在诊断多发性骨髓瘤中的临床应用

目的 探讨多发性骨髓瘤(MM)患者血清轻链(sLC)κ/λ比值及血清蛋白电泳(SPE)在MM诊断中的临床价值.方法 采用速率散射免疫比浊法测定282例MM患者及521例健康对照人群sLC水平并计算κ/λ比值,同时做SPE和免疫固定电泳(IFE).以临床已确诊为MM的患者分型结果为标准,评价sLC比值联合SPE检测对MM...     

Association of anion gap with thyroid dysfunction and nodular goiter in CAPD patients.

OBJECTIVE: To investigate the association of clinical parameters and serum anion gap with thyroid dysfunction and nodular goiter in continuous ambulatory peritoneal dialysis (CAPD) patients. DESIGN: Cross-sectional study. SETTING: Single dialysis unit and outpatient clinic. PATIENTS: This study Included 89 uremic patients on CAPD. Gender ratio was 50 males to 39 females (M/F = 1.28); mean age was 54.8 years. MAIN OUTCOME MEASURES: We investigated the prevalence of nodular goiter and thyroid dysfunction with a 10-MHz high-frequency ultrasound scanner and immunoassay kits. RESULTS: Nodular goiter was detected in 52.8% (47/89) of the CAPD patients. Patients with nodular goiter were older than those without goiter (57.7 vs 51.5 years, p < 0.05). Nodular goiter was found more frequently in females than in males (66.7% vs 44.0%, p < 0.05). Patients with nodular goiter had longer duration of CAPD than patients without goiter (51.6 +/- 42.9 vs 31.0 +/- 28.1 months, p < 0.02). In addition, CAPD patients with goiter had a higher serum anion gap (AG) (16.8 +/- 3.3 vs 14.0 +/- 4.5 mEq/L, p < 0.02) and a lower weekly creatinine clearance (55.9 +/- 12.6 vs 64.6 +/- 21.1 L/week/1.73 m2, p < 0.05) than patients without goiter. As serum AG gradually Increased, significant alteration of thyroid parameters developed In the following sequence: (1) reduction of total T3 level at an AG level of 15, (2) elevation of thyrotropin (TSH) and Increased prevalence of goiter at an AG of 18, and (3) reduction of free T4 and total T4 levels and elevation of TSH, with further increased frequency of goiter at an AG of 20 mEq/L. CONCLUSION: According to this study, age, gender, dialysis duration, serum AG, and weekly creatinine clearance are correlated with prevalence of goiter in CAPD patients. Sequential alteration of thyroid function and Increasing frequency of nodular goiter correlated with higher serum AG. There are two explanations for this correlation: the level of serum AG may be an indirect index of the level of serum goitrogens, and higher serum AG and Increased frequency of nodular goiters might be a reflection of loss of residual renal function. Therefore, thyroid function screening and goiter detection using ultrasound should be considered when examining CAPD patients with progressively elevating serum anion gap.     

Unaccounted for anion in metabolic acidosis during severe sepsis in humans

OBJECTIVE: To quantitate the contribution of lactate, phosphate, urate, total serum proteins, and unidentified anions to the anion gap in patients with severe sepsis. DESIGN: Thirty critically ill patients with evidence of severe sepsis and systemic hypoperfusion were prospectively studied. MEASUREMENTS: The anion gap was calculated as [Na+] + [K+] - [Cl-] - [HCO3]. A corrected anion gap was calculated as the anion gap minus the anionic contribution of lactate, phosphate, urate, and total serum proteins. The corrected anion gap is a marker of unmeasured anion less unmeasured cation concentration. RESULTS: The mean anion gap was 21.8 +/- 1.4 mmol/L and the corrected anion gap was 3.7 +/- 0.8 mmol/L. The mean arterial blood lactate concentration was 5.9 +/- 0.8 mmol/L. The magnitude of the lactate concentration correlated linearly with the anion gap (r2 = .61, lactate = 0.4 anion gap - 3.9, n = 30, p less than .01). The corrected anion gap was greater than 0 in 24 (80%) of 30 patients. The magnitude of the corrected anion gap correlated linearly with the anion gap (r2 = .66, corrected anion gap = 0.5 anion gap - 6.3, n = 30, p less than .01). Since the slope of the regression line for estimating corrected anion gap from anion gap was 0.5, the contribution of unmeasured anions was as important as lactate in determining the anion gap. CONCLUSION: These data indicate that lactic acidosis does not entirely account for the metabolic acidosis during severe sepsis. Furthermore, the increased corrected anion gap suggests the presence of an unidentified anion (or anions) that is (or are) responsible, in large part, for the development of metabolic acidosis in patients with sepsis.     

Simultaneous analysis of serum immunoglobulins in patients with M protein using cellulose acetate membrane isoelectric focusing.

We developed a method for the simultaneous analysis of microheterogeneity of human serum IgG, IgA, IgM, IgD, and IgE, and serum protein pattern using cellulose acetate membrane isoelectric focusing, and analyzed in 11 healthy subjects and 67 patients with M protein (17 cases of multiple myeloma [MM] and 50 cases of monoclonal gammopathy of undetermined significance [MGUS]). Using this method, bands indicating the microheterogeneity of each immunoglobulin could clearly be detected.Among healthy subjects, the detected IgG, IgA, and IgM bands did not vary, but the detected IgE and IgD bands did vary. Therefore, IgA, IgM, and IgG were selected for comparison of serum immunoglobulins in MM and in MGUS. In the IgA-type M protein group, normal IgM and IgG bands were decreased in MM patients compared to MGUS patients, while the M band and other bands were increased in MM patients compared to MGUS patients, but the differences between the two groups were not significant. In the IgG-type M protein group, normal IgM, IgA, and IgG were significantly decreased in MM patients compared to MGUS patients. We examined the changes in electrophoretic pattern in six MM patients and eight MGUS patients with IgA-type M protein after neuraminidase treatment. The width of the M band in MM patients with IgA-type M protein decreased with neuraminidase treatment. On the other hand, the width of the M band in MGUS patients with IgA-type M protein increased with neuraminidase treatment. We concluded that the decrease of the normal immunoglobulins in MM patients with IgG type M protein could be detected by this method, and IgA type of M protein binding sugar chain were different between MM and MGUS patients.     

The predictive power of serum kappa/lambda ratios for discrimination between monoclonal gammopathy of undetermined significance and multiple myeloma.

The predictive power of serum kappa/lambda ratios on initial presentation of immunoglobulin G (IgG) or IgA monoclonal component was studied to differentiate between monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM) patients. The retrospective study involved 145 patients clinically diagnosed with monoclonal gammopathy of undetermined significance or multiple myeloma, who had serum M-protein IgG <35 g/L or IgA <20 g/L at M-protein detection. Serum light chains kappa and lambda were measured by fixed-time nephelometry. Test performance indices, predictive values and likelihood ratios were calculated according to the Weissler recommendation. MM patients were considered as diseased and MGUS patients as non-diseased in order to estimate the performance characteristics of serum kappa/lambda ratios. There was a statistically significant difference in kappa/lambda ratios distribution between both groups of patients, in both M-protein kappa-type (Mann-Whitney U=168, p<0.001) and in M-protein lambda-type (Mann-Whitney U=143, p<0.001). Negative likelihood ratios at threshold levels of 0.6 and 4.2 were 2.17- and 3.32-fold greater, respectively, than positive likelihood ratios, so that the predictive power of a serum kappa/lambda ratio within these limits is better in ruling out (negative predictive power) than ruling in disease (positive predictive power). The post-test characteristics of a serum kappa/lambda ratio interval between 0.6 and 4.2 in discriminating MGUS from MM in our geographic population were: sensitivity 0.96 (0.93-0.99 95% CI); specificity 0.70 (0.63-0.77); positive predictive value 0.68 (0.64-0.73); negative predictive value 0.96 (0.94-0.99); likelihood ratios (+)LR 3.23 (2.68-4.04); and (-)LR 17.16 (11.00-63.00). Thus, serum M-protein with a kappa/lambda ratio between 0.6 and 4.2 increases the posterior probability of MGUS from 0.60 to 0.96 in asymptomatic patients, for whom only monitoring may be suggested when the serum kappa/lambda ratio is within these limits.     

Clinical significance of circulating endothelial adhesion molecules (sE-selectin and sICAM) in untreated multiple myeloma patients

BACKGROUND: The expression of adhesion molecules is important for the interaction of myeloma cells with the bone marrow microenvironment. In the current study, serum soluble adhesion molecules (sICAM-1 and sE-selectin) were measured in untreated multiple myeloma (MM) patients in relation with other markers of disease activity. MATERIALS and METHODS: The study group consisted of 67 patients with MM (classified according to the Durie-Salmon classification) and 15 controls. Interleukin-6 (IL-6), sICAM-1 and sE-selectin concentrations were determined by enzyme-linked immunosorbent assay (ELISA). In addition, the monoclonal protein, erythrocyte sedimentation rate (ESR) and hemoglobin (Hb) concentration were also determined. RESULTS: Serum sICAM-1 level increased significantly at advanced stages of MM and was higher in comparison to controls (p<0.01). sE-selectin increased significantly with advancing stage of the disease, but did not differ from controls. IL-6, ESR and M-component were significantly higher and Hb concentrations lower with advancing stage of disease. There was a positive correlation of IL-6 with sICAM-1 and sE-selectin. CONCLUSIONS: We conclude that serum sICAM-1 differs in multiple myeloma patients from normals and together with sE-selectin increase in parallel to increasing stage of disease, which may reflect a dysregulation and possible involvement of these adhesion molecules in myeloma progression.     

多发性骨髓瘤血清蛋白电泳与免疫学分型

目的对38例多发性骨髓瘤(MM)患者血清蛋白电泳、免疫电泳及免疫球蛋白进行定量分析,以探讨M蛋白、MM型别分布及其在MM中的临床诊断价值。方法血清蛋白和免疫电泳采用全自动琼脂糖凝胶电泳仪进行电泳和扫描;免疫球蛋白定量采用Beckman CS-免疫化学分析仪,以全自动方式进行速率散射比浊分析,测各样本的免疫球蛋白(IgG、IgA、Ig M)含量。结果38例MM全部检出M带,22例MM尿液检出本-周氏蛋白,检出阳性率为57.90%。血清免疫电泳分型IgGκ型14例,占36.84%;IgGλ型10例,占26.32%;IgAκ型5例,占13.16%;IgAλ型6例,占15.79%;Ig Mλ型3例,占7.89%,为最少。结论血清蛋白及免疫电泳能快速、准确地进行MM分型,对MM的诊断、病情判断、治疗及预后评估均具有重要的临床价值。     

Antioxidant defense capacity in scleroderma patients

BACKGROUND: Oxidative stress is associated with scleroderma (systemic sclerosis) and is supposed to favor disease progression by complex effects on the vascular endothelium and on fibroblasts. METHODS: Plasma oxidative process marker, thiobarbituric acid-reactive substances, and several markers of antioxidant defense capacity (plasma total antioxidant activity, serum albumin, uric acid and glutathione, superoxide dismutase and catalase) were evaluated by spectrophotometric methods using blood samples collected from 23 scleroderma patients and 21 healthy controls. RESULTS: In scleroderma patients, thiobarbituric acid-reactive substances levels (mmol/L plasma) were significantly elevated (29.3+/-5.8) compared with healthy controls (16.6+/-3.1, p<0.001). Total antioxidant activity (mmol Trolox/L) was significantly lower in scleroderma patients than in controls (1.29+/-0.13 vs. 1.55+/-0.23, p<0.001), as well as the antioxidant gap (mmol Trolox/L) (0.57+/-0.18 vs. 0.92+/-0.22, p<0.001). Superoxide dismutase activity (IU/g hemoglobin) was markedly decreased in patients as compared with controls (395+/-184 vs. 659+/-211, p<0.001). CONCLUSIONS: Lower plasma total antioxidant activity and plasma antioxidant gap in scleroderma patients show that plasma antioxidant defense is deficient in scleroderma patients. As previous studies on this issue are controversial, the decreased erythrocyte superoxide dismutase activity found in the patients in this study needs further investigation.     

SIRT2 and SIRT3 expression correlates with redox imbalance and advanced clinical stage in patients with multiple myeloma

ObjectivesSirtuins comprise seven family elements (SIRT1-7) involved in various cell signalling pathways comprising cancer inhibition and tumorigenesis. The present study aims to evaluate SIRT2 and SIRT3 gene expression and potential redox reactions in patients with multiple myeloma (MM) at onset and its correlation with disease status, extent and presence of organ damage secondary to myeloma.Design & MethodsTotal RNA was extracted from 17 MM patients and 10 controls to assess gene expression using real-time PCR. The NAD+/NADH ratio as well as the levels of glutathione peroxidase (GPx) and hydrogen peroxide (HP) in peripheral blood mononuclear cells (PBMCs) were determined using established biochemical assays.ResultsSIRT2 and SIRT3 expression is reduced in MM patients compared to healthy controls. Correlational analysis demonstrated that SIRT2 reduction is associated with advanced clinical stage and with more advanced bone lesions than in the remaining patients. SIRT3 expression is correlated with lytic bone lesions. Biochemical analysis indicated an imbalance of oxidative stress biomarkers with low concentrations of the antioxidant enzyme GPx, low amounts of NAD + and higher concentrations of pro-oxidant enzyme HP in PBMCs of MM patients compared to controls. Moreover, MM patients with bone lesions had lower concentrations of NAD + and GPx in PBMCs than patients without signs of bone disease. In addition, MM patients had higher quantities of intracellular HP than controls.ConclusionsOur results demonstrate that SIRT2 and SIRT3 are downregulated in MM and that lower concentrations correlate with an advanced stage of disease and redox imbalance. We conclude that SIRT2 and SIRT3 together with oxidative stress biomarkers, may be useful for improved risk stratification of MM patients.     

用多聚酶链反应技术检测多发性骨髓瘤患者免疫球蛋白重链基因重排的研究

用多聚酶链反应（ＰＣＲ）技术检测了３５例多发性骨髓瘤（ＭＭ）患者的外周血（其中８例同时测骨髓）及１例未定性单克隆丙种球蛋白病（ＭＧＵＳ）患者的外周血和骨髓的免疫球蛋白重链（ＩｇＨ）基因重排。８例ＭＭ的骨髓标本７例阳性。３５例外周血２３例阳性，阳性率为６５．７％。其中ＩＩ、ＩＩＩ期阳性率明显高于Ｉ期；ＩｇＧ型阳性率高于ＩｇＡ及ＩｇＤ型。另１例ＭＧＵＳ患者的外周血及骨髓均未测出ＩｇＨ基因重排。作者认为，ＩｇＨ基因重排的检测对ＭＭ诊断有一定的价值；骨髓标本优于外周血。     

31例轻链型多发性骨髓瘤实验室指标分析

目的探讨实验室检测指标在轻链型多发性骨髓瘤(MM)诊治中的应用价值。方法收集轻链型MM患者31例及对照组60例,对照组为健康者20例(健康组),以及该院诊断的IgGκ型和IgAκ型MM患者各20例。检测免疫球蛋白、血清蛋白电泳、免疫固定电泳、尿素氮(UREA)、肌酐(Cr)、β_2微球蛋白,并进行流式细胞免疫表型检测及骨髓细胞学检查。结果血清蛋白电泳检测到M蛋白的阳性率为29.03%,Durie-Salmon临床分期Ⅱ期与Ⅲ期M蛋白水平差异有统计学意义(P0.05);免疫固定电泳结果均需用琼脂糖凝胶电泳法加做IgD及IgE抗体复查;轻链型MM患者与健康者的IgG、IgA、IgM、UREA、Cr、β_2微球蛋白比较,差异有统计学意义(P0.05);MM细胞形态特征改变显著,细胞抗原表达以CD38、CD138、CD56为主。结论免疫球蛋白定量及免疫固定电泳可作为M蛋白的筛查方法,血清蛋白电泳可用于患者的临床分期和疗效观察;骨髓细胞学检查对轻链型MM起到重要的诊断作用,轻链型MM比其他类型MM有更加严重的肾功能损伤。     

Expansion of Fc receptor-bearing T lymphocytes in patients with immunoglobulin G and immunoglobulin A myeloma.

Lymphocytes obtained from the blood of normal individuals and six patients with newly diagnosed multiple myeloma were separated into T and non-T cell populations by rosette-formation with sheep erythrocytes, and were then assayed for the presence of surface membrane Fc receptors. When compared with normal individuals, four patients with IgG myeloma had a three- to fourfold increase in T cells with IgG receptors (T gamma cells) and two patients with IgA myeloma had a two- to threefold increase in T cells with IgA receptors (T alpha cells). Patients with IgG or IgA myeloma had normal numbers of non-T lymphocytes with surface receptors for IgG and IgA, respectively. The finding that human myeloma is accompanied by elevated numbers of T cells with Fc receptors for the heavy chain class of the myeloma protein: (1) may account for the apparent "monoclonal" lymphocyte population in patients with myeloma; (b) extends to humans similar observations made in mice with secretory plasmacytomas; and (c) is of interest because T cells with Fc receptors are immunoregulatory lymphocytes.