DNA Index in childhood acute lymphoblastic leukaemia: a karyotypic method to validate the flow cytometric measurement

The DNA index (DI) is a prognostic factor in childhood acute lymphoblastic leukemia (ALL). The accuracy of DI measurement is important for treatment stratification: hyperdiploidy with DI ≥ 1.16 is predictive of favorable prognosis whereas hypodiploidy is associated with poor prognosis. The aim of this study was to validate the accuracy of the DI measured by flow cytometry (FCM) by comparison with the karyotype. From samples of 112 childhood ALL, we created a formula to calculate a theoretical DNA index (tDI) based on the blast cell karyotype, taking into account the additional or missing chromosome material of the major clone. FCM DI correlated with tDI calculated from karyotype (R = 0.987) and with modal chromosome number (DI = 0.0202 × Modal NB + 0.0675 and R = 0.984). In three cases a hypodiploid blast cell population was detected by FCM, while only the duplicated clone was identified by the karyotype. The strong correlation between tDI and DI validates the accuracy of FCM quantification, which is technically fast on fresh or frozen samples. If the karyotype is essential to analyze chromosomal abnormalities, FCM provides complementary information in aneuploid ALLs, either by confirming the cytogenetic data or by detecting additional clones not identified when only using cytogenetic data.     

Flow cytometry (FCM) of early radiation response in epidermoid carcinoma of uterine cervix.

DNA flow cytometry (FCM) investigation of tumor specimens before and after 30 Gy 137Cs radiation treatment was performed in 33 cases of epidermoid uterine cervix carcinoma. Distinct differences in the type of FCM response to radiation were seen when the results of DNA index (DI) in diploid and aneuploid tumors and proliferation index (PI) values in diploid tumors from pretreatment and 30 Gy irradiated specimens were compared. We observed partial or total reduction of PI in 12 of 17 diploid and near diploid tumors, and total reduction of the aneuploid population in 14 of 16 aneuploid tumors. No significant correlation was found between the type of FCM response and clinical stage of the disease or the histological degree of differentiation.     

Prognostic significance of flow cytometric DNA analysis in hepatocellular carcinoma: a prospective study of 124 patients

The prognostic significance of the nuclear DNA content of tumors was studied prospectively in 124 patients who underwent hepatic resection for the treatment of hepatocellular carcinoma (HCC). The DNA content was measured by means of flow cytometry (FCM) using fresh or frozen samples. The DNA index (DI) was calculated, and the nuclear DNA content was classified into two types, DNA diploid and DNA aneuploid. The incidence of DNA aneuploid was 55.6%, and the DI ranged from 1.00 to 3.66, with most values falling between 1.00 and 2.00. There. was a significant difference in overall survival between patients with DNA diploid and DNA aneuploid tumors (P < 0.05), with 3-year survival rates being 94.4% and 51.9%, respectively. Among the DNA aneuploid tumor-bearing patients, 55 patients with a high DI (> 1.5) had a worse prognosis than 14 patients with a low DI (≤ 1.5). Of the 98 patients who underwent curative operations, the 43 DNA diploid tumor-bearing patients had more favorable disease-free survival than the 55 DNA aneuploid tumor-bearing patients (P < 0.05, the 3-year disease-free survival rate: 48.4% vs. 0.0%). These results indicate that nuclear DNA content as measured by FCM has prognostic significance for post-operative HCC patients.     

Detection of cytarabine resistance in patients with acute myelogenous leukemia using flow cytometry

Cytarabine (Ara-C) is currently used in the treatment of adult acute myeloid leukemia (AML). To predict the results of induction chemotherapy, it could be useful to detect leukemic cells that are resistant to Ara-C in patients with AML. Using a bromodeoxyuridine/DNA (BrdUrd/DNA) staining method in flow cytometry (FCM), we have developed a cell resistance index to Ara-C (RI). The technique has been applied to 121 bone marrow (BM) samples from patients with de novo AML treated by a regimen containing Ara-C and daunorubicin (DNR). Ninety-seven patients achieved a complete remission (CR), and 24 patients did not and were considered drug-resistant (DR). The BM cells collected at diagnosis were cultured for 48 hours and underwent BrdUrd/DNA analysis. Among 25 patients with no or very low proliferative activity (<3% of cells in S-phase), the proportion of DR patients (nine of 25) was significantly higher than in a second group of 96 patients with detectable proliferative activity (15 of 96) (P < .025). Within this second group, there was a first group of nine patients with high RI values, which included only DR patients; a second group of 63 patients with low RI values, which included 62 CR patients; and a third group of 24 patients with intermediate RI values, which included 19 CR and five DR patients. In view of this series, our results show that it is possible to detect a majority of DR patients treated by Ara-C.     

FCM-DNA测定对急性白血病化疗效果和预后的判断作用

应用流式细胞术(FCM)对41例急性白血病(AL),25例完全缓解期(CR)AL骨髓抽取液进行DNA含量的测定结果表明:初发AL的DNA指数(DI)和增殖期细胞百分率(P％)对预后没有明显影响(P<0.05).而化疗后P%下降,DNA非整倍消失及骨髓白血病细胞减少可作为治疗有效的指标。有效者CR率(81.82%)明显高于无效者(22.22％)(P<0.05).8例复发前1.5～5个月有DNA非整倍体者均出现临床复发.有DNA非整倍体的CR病人复发率明显高于DI正常者(88.89％/31.25％)(P<0.05).对这种病人若采用强烈化疗,可遏止复发,延长CR期.因此FCM-DNA测定是探测CR期AL微小残留病变(MRD)和判断化疗效果的一项简便、可靠的方法。     

A Survey of Regenerating Capacity in Patchy Liver and in Nodular Liver—with Special Reference to DNA Synthesis Estimated by BrdU Labeling Index—

Abstract : Laparoscopically, patchy liver and nodular liver were considered as features of regeneration of liver cells. In this study, the regenerating capacity of liver cells obtained by a liver biopsy from laparoscopically identified patchy liver and nodular liver was estimated by Bromodeoxvuridine (BrdU)-anti-BrdU method. BrdU labeling indices (L. I.), of liver cells in biopsy specimens from 6 normal livers, 12 patchy livers, and from 15 nodular livers were examined using an in-vitro labeling technique. Liver biopsy specimens obtained by a Tru-cut needle were immediately incubated for 45 min. in 0.1% BrdU solution in RPMI 1640 at 37°C under a pressure of 3 atmospheres in a mixture of 95% O₂ and 5% CO₂. Immunohistochemical detection of BrdU was performed by the Avidin-Biotin-Peroxidase Complex (ABC) method. The mean BrdU L. I. (±SE) of normal liver, patchy liver, and of nodular liver was 0.25±0.09%, 1.4±0.2%, and 1.7±0.4% respectively. Among the nodular liver, flat shaped ones showed a low level (0.5±0.1%), in contrast to the high level (2.4±0.7%) in the semispherical nodular liver. The BrdU L. I. of both the patchy liver and the nodular liver was significantly higher than that in the normal liver (p<0.001, p<0.01 respectively). Interesting enough, there was a significant difference (p<0.05) between flat shaped nodular livers and semispherical ones. The semispherical nodular liver demonstrated the highest capacity of DNA synthesis in all the groups examined.     

Plasma Vasopressin in Hereditary Cranial Diabetes Insipidus

ABSTRACT A family comprising 46 members of 4 generations is described; 21 members suffered from incomplete diabetes insipidus (DI) of central origin. The pedigree showed a dominantly transmitted condition. The onset is gradual and starts in early infancy. The clinical symptoms are highly variable and decline in the sixth decade. Plasma vasopressin (AVP) during water deprivation was significantly lower in the DI group than in the controls (4.2±0.5 vs. 10.6±1.7 ng/l) (p<0.01), the difference being more pronounced in the high osmolality range (4.8±0.7 vs. 14.4±3.1 ng/l) (p<0.01). Urine osmolality was lower (241±36 vs. 928±46 mOsm/kg H₂O) (p<0.01) despite higher serum osmolality during water deprivation, rendering the ratio between urine and serum osmolality less than unity compared with >3:1 in the control group (p<0.001). In two affected females, addition of a nonosmotic stimulus caused no increase in plasma AVP. The findings are consistent with a partial defect in the production or release of AVP and not with a dysfunction of the intracranial osmoreceptors. The variable features of incomplete DI indicate that to define the condition by excessive urinary output alone is insufficient. The ratio between urine and serum osmolalities after an appropriate osmotic stimulus together with plasma AVP measurements may be necessary to confirm the diagnosis.     

Sources of variability in the radionuclide angiographic assessment of ejection fraction: A comparison of first-pass and gated equilibrium techniques

Measurements of ejection fractions (EF) determined by first-pass and gated equilibrium radionuclide angiography are widely believed to be equivalent. To compare these measurements in a large group of patients over a wide range of EF values, left ventricular (LV) and right ventricular (RV) EFs at rest were measured in 135 consecutive patients who underwent the 2 methods of radionuclide angiography within 1 hour: first-pass upright with a multi-crystal camera in the anterior projection and gated equilibrium supine with a single-crystal camera in the left anterior oblique projection. The population included 18 normal patients and 117 patients with various cardiac and pulmonary disorders. First-pass and gated equilibrium LVEF correlated well (r = 0.83, p <0.001), but the slope of the regression line was different from unity, with the first-pass values lower than the gated equilibrium values (0.51 ± 0.16 vs 0.56 ± 0.15, p <0.05 [mean ± standard deviation]). Among the 45 patients with a gated equilibrium LVEF of <0.50, the correlation (r = 0.84) was better than that for the 90 patients with a LVEF > 0.50 (r = 0.44, p <0.05). However, in the latter group, the correlation remained good in the 15 patients with cardiomegaly due to aortic or mitral regurgitation (r = 0.80). Inter- and intraobserver error was similar for both methods. In contrast, there was a poor correlation between first-pass and gated equilibrium RVEF, with the first-pass values higher than the gated equilibrium values (0.51 ± 0.11 vs 0.43 ± 0.11, p <0.01). Interobserver error was similar for both the methods, but intraobserver error was better for the first-pass method (p <0.05). Thus, there may be considerable variability in the radionuclide EF at rest in the same patient because of differences in the method of measurement. Caution is suggested when EF values that have been derived using different radionuclide methods are compared.     

Dietary Advice Based on the Glycaemic Index Improves Dietary Profile and Metabolic Control in Type 2 Diabetic Patients

The effect of dietary education incorporating information about the glycaemic index of carbohydrate was tested against standard dietary advice in a randomized controlled study in 51 newly diagnosed patients with Type 2 diabetes treated as out-patients with diet only over a 12-week study period. Outcome was assessed by dietary analysis of 3-day diet diaries, fasting blood glucose, fructosamine, total cholesterol, LDL-cholesterol, HDL-cholesterol, and triglycerides. Dietary analysis indicated that the group who received low glycaemic advice not only had a significantly lower calculated mean diet glycaemic index intake (77 ± 1.1 (SEM) vs 82 ± 1%, p < 0.01) but also had a lower fat intake (25 ± 1 vs 32 ± 2% of total energy day^?1, p < 0.001), a higher carbohydrate intake (49 ± 2% vs 44 ± 1% of total energy day^?1, p < 0.05) and non-starch polysaccharide intake (21 ± 1.5 vs 14 ± 1 g, p < 0.01). There was a significantly greater within-group fall in fructosamine (3.8 ± 0.2 to 3.2 ± 0.2 mmol^?1 vs 3.6 ± 0.2 to 3.6 ± 0.3 mmol^?1, p < 0.05) and cholesterol (6.1 ± 0.3 to 5.4 ± 0.3 mmol^?1 vs. 5.6 + 0.2 to 5.3 ± 0.1 mmol^?1, p < 0.05) in the low glycaemic index group. A significant correlation was detected between the glycaemic index of the diet and the fall in fructosamine (r = 0.54, p < 0.01) and fasting blood glucose (r = 0.41, p < 0.05) which could not be demonstrated with the changes in fat or non-starch polysaccharides content of the diet, irrespective of the original study group, suggesting an independent role of the change in glycaemic index of the carbohydrate in the diet. Subjects given advice to lower the glycaemic index of the carbohydrate in their diet achieve a lower calculated dietary glycaemic intake than those who do not. This appears to have a beneficial effect on the intake of other nutrients and results in an improvement in metabolic control.     

Plasma Endothelin Levels and Blood Pressure in Hemodialysis and in Capd Patients. Effect of Subcutaneous Erythropoietin Replacement Therapy

We investigated plasma endothelin (ET) concentration and blood pressure in 44 patients with end stage renal failure chronically treated with either hemodialysis (n = 24) or continuous ambulatory peritoneal dialysis (CAPD) (n = 20). Half of the subjects were on chronic erythropoietin (r-HuEPO) replacement therapy (30-60 U/kg) subcutaneously, 3 times weekly. The mean plasma ET level of the whole group was about five fold higher than the normal range. Plasma ET concentration and mean blood pressure were higher in hemodialysis than in CAPD patients (33.3 ± 2.1 vs 24.8 ± 1.2 pg/ml, p < 0.01, and 101 ± 2.4 vs 91 ± 3 mmHg, p < 0.025). There was a significant correlation between plasma ET levels and systolic blood pressures in both groups (r = 0.45, p < 0.05). Patients (hemodialysis and CAPD) receiving subcutaneous r-HuEPO had higher mean blood pressure (99 f 3 vs 85 Ifi 4 mmHg, p < 0.01), while their plasma ET levels were similar to untreated patients independent of the dialysis mode. However, a statistically significant correlation between plasma ET and systolic blood pressure was present only in the r-HuEPO treated group (r = 0.46, p < 0.05 vs r = 0.29, N.S., for the untreated group). These results show that plasma ET levels are markedly increased on both dialysis mode, but the values are lower in CAPD patients. Plasma ET concentrations significantly correlated with systolic blood pressures in the whole group of patients, and also in those receiving r-HuEPO replacement therapy.     

Relationship of AgNOR counts and nuclear DNA content to survival in patients with parathyroid carcinoma

The aim of this study was to evaluate the usefulness of DNA flow cytometry to determine tumor nuclear DNA index (DI), and nucleolar organizer region protein counts visualized by the argyrophil (AgNOR) technique, in confirming diagnosis and predicting clinical outcome of patients with parathyroid carcinoma (PC). We reviewed paraffin-embedded tissue sections, from 15 patients (median age 63 years, range 30-68 years) with PC who died of the disease, which were randomly compared with tissue sections from 15 age- and sex-matched patients with parathyroid adenoma (PA). The proliferative activity in parathyroid tumours as detected by DI and AgNOR counts was evaluated in all specimens. Both DI (1.37 +/- 0.33 vs 1.0 +/- 0.1) and AgNOR (3.01 +/- 0.31 vs 1.54 +/- 0.35) counts were higher (P < 0.001) (Student's t-test) in patients with PC than in those with PA. Diploid (DI = 1), aneuploid (DI > 1) and hypoploid (DI < 1) neoplasms were found in 11 (PC = 4, PA = 7), 14 (PC = 11, PA = 3) and five (PC = 0, PA = 5) patients respectively. The average postoperative survival in patients with PC was 46.9 +/- 37.4 months (range 21-146 months). The survivals of patients with aneuploid (n = 11) and diploid (n = 4) PC were 74.0 +/- 58.1 and 34.1 +/- 18.4 months (P=0.21) respectively. There was a significant relationship between DI and AgNOR counts (R=0.69, P < 0.01), but no correlation was found between survival and both DI (Rho = 0.17, P = 0.55) and AgNOR counts (Rho = 0.26, P = 0.35). Moreover, there was no correlation (P = NS) between the main preoperative biochemical parameters and survival. In conclusion, DI and AgNOR are useful in confirming the diagnosis of PC, but they are of little value in predicting the clinical outcome of patients with PC.     

Evaluation of Biopsy Classification for Rejection: Relation to Detection of Myocardial Damage by Monoclonal Antimyosin Antibody Imaging

Objectives. This study sought to compare the histologic grades of rejection in endomyocardial biopsy specimens with the global estimate of myocardial transplant-related cardiac damage detected by myocardial uptake of monoclonal antimyosin antibodies.Background. The diagnosis and treatment of acute cardiac allograft rejection is based on the interpretation of endomyocardial biopsies. Because allograft rejection is a multifocal process and biopsy is obtained from a small area of the right ventricle, sampling error may occur. Global assessment of myocardial damage associated with graft rejection is now possible with the use of antimyosin scintigraphy. The present study was undertaken to compare the histologic grades of rejection in endomyocardial biopsy specimens with the global assessment of transplant-related myocardial damage detected by antimyosin scintigraphy.Methods. Biopsies (n = 395) from 112 patients were independently interpreted by three pathologists in a blinded manner according to the original Stanford four-grade (normal, mild, moderate and severe) and the current International Society of Heart and Lung Transplantation (ISHLT) seven-grade (0, 1A, 1B, 2, 3A, 3B and 4) classifications. The results were correlated with 395 antimyosin studies performed at the time of the biopsies. The heart/lung ratio of antimyosin antibody uptake was used to assess the severity of myocardial damage.Results. In the Stanford biopsy grade classification, significantly higher antimyosin uptake, indicating increasing degrees of myocardial damage, were associated with normal (1.78 ± 0.26), mild (1.88 ± 0.31) and moderate (1.95 ± 0.38) biopsy classifications for rejection (p < 0.01). In the ISHLT classification, significant differences were detected only for antimyosin uptake associated with grades 0 (1.77 ± 0.26) and 3A (1.98 ± 0.39) but not for intermediate scores (1A, 1B and 2). In view of the similar intensity of antibody uptake among the various grades, ISHLT biopsy scores were regrouped: normal biopsies in grade A; 1A and 1B as grade B; and 2 and 3A as grade C. Antimyosin uptake in grades A, B and C was 1.78 ± 0.26, 1.88 ± 0.31, 1.95 ± 0.38, respectively (p < 0.01).Conclusions. The current ISHLT seven-grade scoring system does not reflect the progressive severity of myocardial damage associated with heart transplant rejection. Because myocardial damage constitutes the basis of treatment for allograft rejection, there is a need to reevaluate the ISHLT grading system, given its importance for multicenter trials.     

流式细胞仪DNA倍体测定鉴别良恶性胸腔积液的诊断价值

目的通过流式细胞仪（FCM）技术测定胸水脱落细胞细胞核的脱氧核糖核酸（DNA）含量,并将此方法与癌胚抗原（CEA）相比较。方法病例选自2004～2006年5月,本院住院患者,留取新鲜的胸腔积液标本,用流式细胞仪检测细胞核DNA倍体,分析细胞的增殖能力。结果在恶性胸腔积液组中70％的标本可以检测到DNA异倍体,在良性胸腔积液中90％的标本表现为两倍体。结论流式细胞仪DNA倍体测定,与CEA联合测定可以提高恶性胸腔积液的检出率。     

Prolactin and thyroid hormones in patients with systemic sclerosis: correlations with disease manifestations and activity

Summary. Objective: To determine serum levels of prolactin (PRL) and thyroid hormones and to investigate the correlation between these hormones and different disease manifestations in patients with systemic sclerosis (SSc). Methods: Twenty four patients with SSc (23 women, mean age 37.7±12.7) were subjected to thyroid hormones assessment. Prolactin (PRL) was assessed in 23female patients. The patients were evaluated regarding different disease manifestations. Fifteen normal female volunteers were involved as controls. Results: Serum levels of PRL in all patients was significantly higher than controls (16.75±9.06 for patients vs. 11.6±4.5 for controls with p<0.001). Eight patients out of 23 (34.8%) showed hyperprolactinemia. In patients with diffuse SSc (dSSc), PRL levels showed significant correlation with the rate of skin tethering (r=+0.72, p<0.01) and abnormal left ventricular filling pattern (↓E/A ratio), i.e., occurrence of diastolic dysfunction (r=+0.65, p<0.05). Hyperprolactinemia in all patients correlated significantly with disease duration (r=–0.42, p<0.05).     Mean serum levels of free thyroxin (FT4) in all patients were significantly lower than the control group(7.46±2.7 for patients vs 10.47±2.5 with p<0.001). Eight out of all 24 patients (33.3%) showed hypothyroidism. In groupA (duration<3years); FT4 levels correlated significantly with Dlco% (r=+0.90, p<0.01). While in groupB (duration>3years), T4 hypothyroidism correlated significantly with hand joint restriction of motion (r=+0.66, p<0.01).    Serum levels of triiodothironine (FT3) in all patients were nonsignificantly lower than the control group (4.8±2.3 for patients vs 5.3±1.9 for controls, P=NS). Three patients out of 24 (12.5%) showed T3 thyrotoxicosis. Serum levels of T3 correlated significantly with liver enzyme elevations (r=+0.46, p<0.05) and ESR (r=+0.41, p<0.05). Conclusion: This study demonstrates the close association between PRL or thyroid hormones and some organ involvement in SSc. Correspondence to Amira A. Shahin     

Cytogenetic analysis of gallbladder neoplasms using fluorescence in situ hybridization (FISH)

To characterize the numerical chromosome aberrations in gallbladder neoplasms, we examined surgically resected tissues using fluorescence in situ hybridization. The aberrations in 15 specimens of adenocarcinomas and 2 adenomas were compared with those in 4 samples of adenomyomatosis and 17 samples of normal epithelium. We calculated the frequency of aneusomy and determined the chromosome indexes (mean number of chromosomes per nucleus) of chromosomes 17 and 18. The pattern of DNA ploidy was analyzed by flow cytometry. In normal epithelium, adenomyomatosis and adenomas, DNA aneuploidy was not observed, while 13 (87%) carcinomas showed DNA aneuploidy, including 2 specimens with multiploidy. No numerical aberrations were observed in normal epithelium and adenomyomatosis. A numerical gain of chromosome 17 was observed in a single adenoma and in 10 (66%) carcinomas. A numerical gain of chromosome 18 was observed in 6 (40%) carcinomas, but not in other tissues. The chromosome index of chromosome 17 was significantly higher in adenomas and carcinomas (2.45±0.60 and 2.29±0.14, respectively) compared with normal epithelium. Our cytogenetic findings did not correlate with any histopathologic features of carcinomas. Our results indicated that the gains of chromosome 17 and 18 represented early chromosomal alterations in gallbladder neoplasms and were maintained in advanced carcinomas.     

Hyponatremia in systemic lupus erythematosus patients: Relation to disease activity and fatigue

Aim of the workTo investigate the relation of hyponatremia to disease activity and fatigue in systemic lupus erythematosus (SLE) patients.Patients and methodsThe present study included 30 SLE patients with hyponatremia and 70 with normal serum sodium (Na) level. SLE disease activity index and the Functional Assessment of Chronic Illness Therapy (FACIT)-Fatigue score were assessed.ResultsThe gender, age and disease duration of SLE patients with hyponatremia (26 females and 4 males; mean age 37.8 ± 9.3 years, 10.9 ± 5.1 years) were comparable with those without (60 females and 10 males; 36.9 ± 11.8 years, 11.1 ± 6.02; p = 0.71 and p = 0.9 respectively). SLE patients with hyponatremia showed significantly increased SLEDAI (14.2 ± 3.85 vs 3.86 ± 3.59; p < 0.001), ESR (55.2 ± 18.2 vs 14.70 ± 3.5 mm/1st h; p < 0.001) and CRP (30.97 ± 4.4 vs 5.17 ± 2.6 mg/dl; p < 0.001) and lower FACIT-Fatigue (21.99 ± 2.1 vs 35.87 ± 4.81; p < 0.001) compared to patients without. Serum Na levels significantly correlated with the FACIT-fatigue score (r = 0.99, p < 0.01), platelets (r = 0.22; p = 0.03), white blood cells count (r = 0.31, p < 0.001) and inversely with SLEDAI (r = ?0.27, p = 0.01), ESR (r = ?0.71; p < 0.001), CRP (r = ?0.86, p < 0.001), anti-ds-DNA (r = 0.54, p < 0.001), C3 (r = ?0.29, p = 0.01) and C4 (r = ?0.2, p = 0.04). On regression, CRP (β = 0.3), SLEDAI (β = 0.28) and consumed C4 (β = ?0.07) were significant independent risk factors for hyponatremia (p < 0.0001, p = 0.0005, p = 0.02 respectively). The optimal cut-off values to predict hyponatremia was a SLEDAI score ≥11 (90% sensitivity and 96% specificity), and ESR ≥ 17.5 mm/1st h (100% sensitivity and 80% specificity) and a CRP of ≥10.5 mg/dl (100% sensitivity and 97% specificity).ConclusionHyponatremia in SLE patients is associated with higher disease activity and more perceived fatigue. Hyponatremia could reflect severe inflammation and could be considered as one of the predisposing factors of fatigue.     

Complement deposition on renal histopathology of patients with diabetic nephropathy

AimsAs the potential role of the complement system in diabetic nephropathy (DN) is increasingly reported, this study aimed to investigate C1q and C3c deposition as seen on renal histopathology, as well as its association with clinical and pathological parameters, in DN patients.MethodsRenal biopsy specimens from 161 DN patients were investigated using direct immunofluorescence, light, and electron microscopy. For direct immunofluorescence, staining for C1q and C3c on fresh-frozen renal tissue was performed immediately after biopsy. Complement deposition was defined as the presence of C1q or C3c of at least 1 + on a 0–4 + Scale. The association between complement deposition and clinicopathological data was also analyzed.ResultsOn direct immunofluorescence microscopy, C1q and C3c were detected in specimens from 44/161 (27.3%) and 89/161 (55.3%) patients, respectively. Regarding clinical data, patients with C1q deposition had a significantly higher level of urinary protein (7.25 ± 4.20 g/24 h vs. 4.97 ± 3.76 g/24 h; P < 0.01) and significantly lower estimated glomerular filtration rate (eGFR; 34.16 ± 25.21 mL/min/1.73 m² vs. 51.17 ± 31.56 mL/min/1.73 m², respectively; P < 0.01), whereas patients with vs. without C3c deposition had a significantly lower eGFR (40.09 ± 27.97 mL/min/1.73 m² vs. 54.48 ± 32.49 mL/min/1.73 m², respectively; P < 0.01). On renal histopathology, patients with C1q deposition had significantly higher Scores for interstitial fibrosis and tubular atrophy (IFTA), interstitial inflammation and vascular lesions (P < 0.01, P < 0.05 and P < 0.05, respectively), whereas patients with C3c deposition had significantly higher IFTA Scores and proportions of global sclerosis (P < 0.01 and P < 0.01, respectively).ConclusionComplement deposition of C1q and C3c on renal histopathology is associated with more severe kidney damage in patients with DN.     

Increased nitric oxide production accompanied by the up-regulation of inducible nitric oxide synthase in vascular endothelium from patients with systemic lupus erythematosus

Objective. To investigate whether systemic lupus erythematosus (SLE) is accompanied by increased serum nitrite levels, whether active compared with inactive disease is associated with greater nitric oxide (NO) production, and whether endothelial cells or keratinocytes serve as cellular sources of NO by virtue of their increased expression of either constitutive nitric oxide synthase (cNOS) or inducible NOS (iNOS). Methods. Fifty-one serum samples (46 from patients with SLE) were analyzed for NO production by measuring nitrite levels in a calorimetric assay. Skin biopsy samples from 21 SLE patients and 11 healthy volunteers were evaluated immunohistochemically, using monoclonal antibodies, for endothelial cell and keratinocyte cNOS and iNOS expression. Results. Serum nitrite levels were significantly elevated in the 46 patients with SLE (mean ± SEM 37 ± 6 μM/liter) compared with controls (15 ± 7 μM/liter; P < 0.01), and were elevated in patients with active SLE compared with those with inactive disease (46 ± 7 μM/liter versus 30 ± 7 μM/liter; P < 0.01). Serum nitrite levels correlated with disease activity (r = 0.47, P = 0.04) and with levels of antibodies to doublestranded DNA (r = 0.35, P = 0.02). Endothelial cell expression of iNOS in SLE patients (mean ± SEM score 1.5 ± 0.2) was significantly greater compared with controls (0.6 ± 0.2; P < 0.01), and higher in patients with active disease compared with those with inactive SLE (1.7 ± 0.2 versus 1.2 ± 0.2; P < 0.01). Keratinocyte expression of iNOS was also significantly elevated in SLE patients (0.9 ± 0.1) compared with controls (0.4 ± 0.1; P < 0.001). With regard to expression of cNOS, there were no differences between patients with active SLE, those with inactive SLE, and normal controls in either the vascular endothelium or the keratinocytes. Conclusion. NO production is increased in patients with SLE, and 2 potential sources of excessive NO are activated endothelial cells and keratinocytes via up-regulated iNOS.     

Biological characteristics of HCC by ultrasound-guided aspiration biopsy and its clinical application

AIM: To probe the pathological biological characteristics of hepatocellular carcinoma (HCC) by the ultrasound-guided aspiration biopsy and assess the clinical application value of this method.METHODS: The biopsy and DNA analysis by flow cytometry (FCM) were taken in 46 cases with HCC nodules, including 26 cases and 20 cases with nodules ≤3 cm and ＞3 cm in diameters respectively, and 12 cases with intrahepatic benign hyperplastic nodules. They were taken in 22 cases of 46cases with HCC before and after the therapy. Fine-needles and automatic histological incised biopsy needles were used.The fresh biopsy tissue was produced into the single cell suspension, which was sent for DNA detection and ratio analysis of cell period. The ratio of each DNA period of cell proliferation of each group was calculated and compared with each other. The DNA aneuploid (AN) and apoptosis cell peak were observed and their percentages were calculated.RESULTS: The ratios of S and G2/M periods of DNA, which reflect cell hyperproliferation, in the group with HCC tumors ＞3 cm in diameter were markedly higher than those of the group with HCC nodules ≤3 cm in diameter and the group with the benign hyperplastic nodules (P＜0.01 except A:B of S period, P＜0.05). The ratios of the middle group were also apparently higher than those of the latter group (P＜0.01).The ratio of DNA AN of 46 cases with HCC nodules was 34.8 % (16/46). None of the cases with the intrahepatic hyperplastic nodules appeared AN. The DNA AN appeared more apparently with the growth of the tumors. The AN ratio of the group with tumors ＞3 cm in diameter was 55 %(11/20), markedly higher than that of the group with tumors ≤3 cm in diameter which was 19.2 % (5/26) (P＜0.01). The FCM DNA analysis of 22 specimens of hepatic carcinoma tissue before therapy showed that the aneuploid peaks appeared in 5 cases (22.7 %). The ratio of G1 period rose after therapy while the S period and G2/M ratios fell (P＜0.01).The aneuploid peak disappeared in the 5 cases after the therapy, while the apoptosis peaks in 12 cases (54.5 %)appeared.CONCLUSION: Addition to supply the information of the pathological morphology of the tumor, the ultrasound-guided fine-needle aspiration tissue could be sent for FCM DNA analysis to comprehend its pathological biological characteristics. This can not only provide the clinic the reliable information about the occurrence, development,diagnosis, curative effect and prognosis of tumors but also supply biological information for clinic to choose therapeutic schemes.