DNA ploidy,proliferative activity,and concanavalin A reactivity in breast cancer

Paraffin-embedded primary tumor specimens from 48 patients with breast cancer were examined for DNA ploidy, S-phase fraction (SPF), and concanavalin A (Con A) reactivity. The results were correlated with clinicopathological prognostic factors, including patients' age and menopausal status, stage of disease, nuclear grade, and size of the primary tumor. There were no associations among ploidy, SPF, Con A reactivity, and menopausal status, stage of disease, or size of the primary tumor. However, among patients who were 50 years or older, 81 % had diploid tumors and 73% had good reactivity (3+ or better staining score) with Con A. In contrast, among patients who were younger than 50 years, 45% had diploid tumors (P < 0.05) and 21% had good Con A reactivity (P < 0.05). Seven of 19 (37%) poorly differentiated tumors and 7 of 9 (78%) moderately differentiated tumors had good reactivity with Con A (P < 0.05). Reactivity of tumor cells with Con A in primary breast cancer tissues deserves further evaluation as a potential biomarker of prognosis. © 1994 Wiley-Liss, Inc.     

Identification of Cryptococcus antigen in human immunodeficiency virus‐positive Turkish patients by using the Dynamiker® lateral flow assay

Cryptococcus neoformans causes life‐threatening meningoencephalitis, particularly in human immunodeficiency virus (HIV)‐positive individuals with low CD4 levels (<100 cells/μL). Although the burden of cryptococcal meningoencephalitis (CM) in Turkey is low (0.13 cases per 100 000 persons), asymptomatic individuals at risk of cryptococcosis should be screened for antigenemia to prevent the disease and/or promote early CM diagnosis. A lateral flow assay (LFA) is used to detect Cryptococcus antigen (CrAg) in cerebrospinal fluid and serum. We determined Cryptococcus antigenemia prevalence in serum samples of HIV‐positive and HIV‐negative adult patients by using Dynamiker® CrAg‐LFA, a point‐of‐care dipstick test. Patients’ demographic data, CD4 count, HIV‐RNA levels and anti‐retroviral therapy status were recorded. CrAg was detected in 28 (11%) of 254 HIV‐positive patients screened but not in 100 HIV‐negative control individuals; a significant difference was observed in the CrAg‐LFA positivity rate between HIV‐positive and HIV‐negative groups (x² = 11.970; P < .05). In CrAg‐positive patients, the median CD4 level was 666 cells/μL (115‐1344 cells/μL), with a median viral load of 23 copies/mL (0‐3.69 × 10⁶ copies/mL). In HIV‐positive CrAg‐negative patients, the median CD4 level was 633 cells/μL (31‐2953 cells/μL) and the median viral load was 12 copies/mL (0‐1.95 × 10⁶ copies/mL; P > .05). Results indicate that HIV‐positive patients with both low (<200 cells/μL) and high (>200 cells/μL) CD4 counts should be screened for asymptomatic cryptococcal antigenemia. HIV‐associated asymptomatic cryptococcosis is not uncommon in Turkey, which warrants systematic screening. Updated strategies for CM prevention among HIV‐positive patients should be used even in non‐endemic countries.     

Expressions of hypoxia-inducible factor-1α and hexokinase-II in gastric adenocarcinoma: the impact on prognosis and correlation to clinicopathologic features

The impact of hypoxia-inducible factor (HIF)-1α and hexokinase-II (HK-II) expression on prognosis of gastric adenocarcinoma patients has not been clearly established. We identified all patients in Cancer Center of Sun Yat-Sen University who were diagnosed as gastric adenocarcinoma and underwent radical gastrectomy between January 1999 and December 2001. We used immunohistochemistry to determine the expressions of HIF-1α protein and HK-II in the surgical sections. We identified 188 patients with gastric adenocarcinoma for the final analysis. The positive rate of HIF-1α and HK-II were 110/188 (54.6%) and 40/188 (21.3%), respectively. Both HIF-1α and HK-II were all positively correlated with tumor size, lower differentiation, and tumor stage. Univariate analysis showed that advanced tumor stages (P < 0.001), tumor size (P = 0.003), HIF-1α expression (P < 0.001), and HK-II expression (P < 0.001) were all significantly associated with shorter survival. The multivariate Cox analysis revealed that tumor stage (P < 0.001), HIF-1α expression (P < 0.001), and HK-II expression (P = 0.002) remained independent prognostic variables for survival. In addition, there was a positive correlation of HIF-1α protein expression and HK-II (P = 0.022). Both HIF-1α and HK-II were overexpressed in gastric adenocarcinoma. The multivariate Cox analysis revealed that both of them were independent factors on survival of gastric adenocarcinoma patients.     

Expression of vascular endothelial growth factor (VEGF) and its two receptors in diffusely infiltrating astrocytomas and relationship to proliferative activity of tumor cells

We studied the relationships among vascular endothelial growth factor (VEGF), its receptors (Flt1 and Flk1), and MIB-1. Their expression in 47 diffusely infiltrating astrocytomas obtained at surgery or autopsy was investigated by the ABC method and analyzed quantitatively. The positive rate of VEGF in tumor cells was higher than that in endothelial cells, and Flk1 was lower in tumor cells (P<0.01, 0.01), whereas Flt1 in both tumor cells and endothelial cells was found at similar levels (P>0.05). In tumor cells, VEGF became high with increased histological grades (P<0.01), whereas both Flt1 and Flk1 were higher in grade 4 than in grades 2 and 3 (P<0.01, 0.05). VEGF, Flt1, and Flk1 in endothelial cells were also highly expressed in grade 4 (P<0.01). The distribution of MIB-1-positive nuclei in grade 4 was similar to VEGF, and the percent of positivity from grade 2 to grade 4 also increased (P<0.01). There was a linear positive correlation between VEGF and both Flt1 and Flk1 in both tumor cells and endothelial cells (P<0.01). So was the percent of positivity with VEGF, Flt1, and Flk1 in tumor cells and endothelial cells (P<0.01). The experiment suggests that VEGF may act as a growth factor for both endothelial cells and tumor cells. VEGF, Flt1, and Flk1 can be considered as indicators of the malignancy potential of diffusely infiltrating astrocytomas. The expression of VEGF and the two receptors may be affected by the proliferative activity of tumor cells.     

FLOW CYTOM ETRIC ANALYSIS OF CELLULAR DNA CONTENT IN EPITHELIAL OVARIAN TUMOR

The DNA content of tumor all was analyzed by flow cytometry on parafflnembedded specimens in 73 patients with epithelial ovarian tumor, and its clinical significance was evaluated. One of the 5 benign (20%), 2 of the 11 borderline (18.18%), and 30 of the 57 malignant (52. 63%) tumors were aneuplold. The occurrence rate of aneuploidy In malignant tumors was higher than In benign and borderline tumors ( P < 0. 05 ). Furthermore, aneuploidy was more frequently In the advanced stages (Ⅲ -Ⅳ ) (77. 7%) than in the early stages (Ⅰ - Ⅱ ) (9. 5%) (P<0. 005). The occurrence rate of DNA aneuploidy was higher in patients associated with ascites and the residual tumor≥.2 cm. Patients with aneuploid tumors had more of ten ascites (P<0. 005) and residual tumor size≥2cm (P< 0.005). There was no apparent correlation between the DNA ptoidy and the histologic grade, histologic type of the tumors. G0/G1 cell proportion of DNA diplold tumors in advanced carcinoma (64. 6%) was less than those of early stage carcinoma (     

Recombinant Apoptin gene retrovirus induces apoptosis in human breast cancer cells 435

Objective  To construct recombinant Apoptin gene (vp3) retrovirus pLVP3 and to study its apoptosis inducing effect on human breast cancer cells 435 as well as to discuss its mechanism in vitro and in vivo. Methods   vp3 gene was cloned and recombinated into retrovirus vector pLP-LNCX-VP3 (pLVP3) at loxP site, which was transformed into package cell line PT67 and then into NIH3T3 cells for titer assay. The human breast cancer cell line 435 was infected with retrovirus pLVP3, and then MTT assay and Western blotting were adopted to detect cellular proliferation and Apoptin protein expression. Forty-eight hours after infection flow cytometry (FCM) was used for apoptosis detection and Surface Enhanced Laser Desorption Ionization-Time of Flight-Mass Spectrometry (SELDI-TOF-MS) was used for protein profile assay. Nude mice model of human breast cancer cells 435 was set up to observe the tumor inhibiting rates of pLVP3, and TUNEL assay was used to detect tumor apoptosis as well as real-time PCR for vp3 gene expression. Results  Recombinant plasmid pLVP3 was successfully constructed. Virus titer reached to 5×10⁸ pfu/ml in the PT67 culture supernatant. Forty-eight hours after infection, cellular inhibition rate was 65.1% in MTT assay, higher than that in blank control (P<0.05) and Apoptin protein expressed more in test group in Western blotting. FCM assay showed apoptotic peaks with a percentage of 15.42%. SELDI-TOF-MS findings suggested that two protein peaks, M_2544.1+H and M_3712.4+H, were statistically different between infection group and control group (P<0.05). The tumor inhibition rates in pLVP3 group were 65.52% and 68.23%, much higher than that of control group (t=4.06, P<0.01). TUNEL assay findings showed that positive yellow stains were seen in pLVP3 retrovirus group and 5-FU positive control group without difference (t1=1.05, t2=0.84, P>0.05). Conclusion  The experiment demonstrated that vp3 could induce apoptosis in tumor cells in vivo and in vitro, which laid a basis for further study on molecular mechanism of tumor cell apoptosis induced by Apoptin and provided valuable reference for tumor gene therapy. Contributed equally to this work.     

Primary Tumor Location Is a Useful Predictor for Lymph Node Metastasis and Prognosis in Lung Adenocarcinoma

Introduction

We examined the value of tumor location in predicting the clinicopathologic features, survival, and metastases of pulmonary adenocarcinoma.

孤立性肺腺癌血管生成与血流模式初步研究:影像-病理对照

Objective:To investigate the correlations of vascular endothelial growth factor (VEGF)-positive tumor anglogenesis and the quantifiable parameters of blood flow pattern derived with dynamic CT in solitary bronchogenic adenocarcinoma.Methods:30 patients with VEGF-positive bronchogenic adenocarcinomas (diameter≤4 cm) underwent multi-location dynamic contrast matedal..enhanced (nonionic contrast material was administrated via the antecubital vein at a rate of 4 mL/sec by using an autoinjector) serial CT.The quantifiable parameters (Perfusion,peak height,ratio of peak height of the bronchogenic adenocarcinoma to that of the aorta and mean transit time) of blood flow pattern derived with dynamic CT in solitary bronchogenic adenocarcinoma were compared with microvessel densities (MVDs) and VEGF expression by immunohistochemistry.Results:Peak height of VEGF-positive bronchogenic adenocarcinoma was 36.06 HU 13.57 HU,bronchogenic adenocarcinoma-to-aorta ratio 14.25%±4.92,and perfusion value 29.66±5.60 mL/min/100 g,mean transit time 14.86 s±5.84 s,and MVD 70.15±20.03.Each of peak height,ratio of peak height of the bronchogenic adenocarcinoma to that of the aorta and perfusion correlated positively with MVD (r=0.781,P＜0.0001;r=0.688,P＜0.0001;r=0.716,P＜0.0001;respectively).No significant correlation was found between mean transit time and MVD (r=0.260,P=0.200＞0.05).Conclusion:Perfusion,peak height and ratio of peak height of the bronchogenic adenocarcinoma to that of the aorta reflect MVD in VEGF-pesitive bronchogenic adenocarcinoma.Perfusion,peak height and ratio of peak height of the bronchogenic adenocarcinoma to that of the aorta derived with dynamic CT might be index for VEGF-related tumor angiogenesis in bronchogenic adenocercinoma.     

Increased NDRG1 Expression is Associated with Advanced T Stages and Poor Vascularization in Non-small Cell Lung Cancer

N-myc downstream regulated gene 1 (NDRG1) is a member of the N-myc downstream regulated gene family which belongs to the alpha/beta hydrolase superfamily. Earlier studies have shown its association with inhibition of tumor metastasis. However, its function in malignant tumors is not fully enunciated. Recently there was increasing evidence that NDRG1 is involved in stress responses. In the current study, we examined the expression of NDRG1 and its correlation with clinicopathological factors and microvessel density (MVD) in non-small cell lung cancer (NSCLC) using immunohistochemistry (IHC). NDRG1 expression in NSCLC (71/115, 61.7%) was higher than that in normal lung tissues (32/115, 27.8%) (p < 0.05). NDRG1 expression in NSCLC cells was found in cytoplasm (63/115, 54.8%), nuclear (24/115, 20.9%) and cell membrane (13/115, 11.3%). NDRG1 expression in NSCLC with advanced T stages (T2-4) (63/84, 75.0%) was significantly higher than that with T1 stage (8/31, 25.8%) (P < 0.05). No other clinicopathological factors including lymph node metastasis were found to be associated with NDRG1 expression (p > 0.05). Moreover increased NDRG1 expression was associated with lower MVD in NSCLC (P < 0.05). MVD in adenocarcinoma (33.4 ± 8.4/HP) was significantly higher than that in squamous cell carcinoma (SCC) (19.3 ± 8.1/HP) (P < 0.05). No other clinicopathological factors were associated with MVD in NSCLC (p > 0.05). The present findings indicate an increase of NDRG1 expression with the progress of tumour extent which may be due to unbalanced tumor oxygenation on account of poor vascularization in NSCLC.     

The relationship between the erythrocyte polyamine levels and the different types of leukemia

In a study on erythrocyte polyamine levels in 20 leukemia patients and 18 healthy individuals, following results were obtained. Putrescine(Pu) in erythrocytes was nearly undetectable but in 2 AMoL patients; Spermidine(Spd) level in ALL was higher than the control volue (P<0.05), in AML (M₁, M₂) and APL the Spd level was in the range of control group (P>0.1), while that in AMoL and AMMoL was considerably lower as compared to control (P<0.001); Spermine (Spm) level in all types of leukemia was increased (P<0.001); Spd/Spm ratio was significantly decreased, showing 1.525 in control group, 0.938 in ALL (P<0.025), 0.779 in AML (M₁, M₂)P<0.01), 0.319 in APL (P<0.001) and 0.296 in AMoL & AMMoL (P<0.001), and the differences between the Spd/Spm ratio in AMoL and AMMoL and in ALL were noted (P<0.05). These results suggested that determination of erythrocyte polyamines in leukemia patients may be helpful in diagnosis, differential diagnosis and prognosis.     

中国启东地区肝癌细胞凋亡相关基因的克隆、表达及其与肝癌相关性研究

Objective To explore the molecular basis of hepatocarcinogenesis by cloning and expressing a novel liver cancer apoptosis-related gene. Methods With homologous screening and RT-PCR, we had cloned an apoptosis-related gene APG from liver cancer cells, compared its expression in hepatocellular carcinoma (HCC) tissue, and paracarcinoma tissue, and analyzed its sequence from these tissues. The association of APG gene expression with HCC was investigated. Results A new gene APG was cloned with a full-length cDNA of 563 bp. Sequencing analysis showed heterogeneity of APG gene from hepatocarcinoma tissue and from paracarcinoma tissue. Among 50 cases of liver cancer, APG gene, expressions were down-regulated in 42 cases (84%), while up-regulated in 8 cases (16%,P<0.01). Its expression was also found to be associated with tumor size (P<0.05), HbsAg level (P<0.01), degree of tumor differentiation (P<0.01), invasion of the tumor capsule (P<0.01), clinical stage (P<0.01), tumor embolism (P<0.01), paracarcinoma tumor satellites (P<0.01), Ki-67 protein expression (P<0.05), p53 protein expression (P<0.01) and apoptosis (P<0.05). There is no association between the gene expression, gender and AFP (P>0.05). Conclusion APC is an appoptosis-related gene and down-regulated in HCC. Its expression is associated with many clinical and pathologic features of HCC, suggesting that APC gene is probably involved in the tumorigenesis of HCC.     

Rosiglitazone and imidapril alone or in combination alleviate muscle and adipose depletion in a murine cancer cachexia model

Rosiglitazone (RGZ) and imidapril improve cancer cachexia via different mechanisms. Therefore, we hypothesized that combination therapy of RGZ+imidapril would further attenuate cancer cachexia in vivo. After injection with colon-26 adenocarcinoma for 9 days, BALB/c mice were randomly divided into the following four treatment groups for 7 days (n?=?8 per group): (1) placebo, (2) RGZ, (3) imidapril, and (4) RGZ+imidapril. Eight healthy control animals were also assessed. Body weight, tumor volume, gastrocnemius muscle and epididymal adipose mass, serum metabolic markers and cytokines, and the expression of nuclear factor-κB and two E3 ubiquitin ligases, atrogin-1 and MuRF-1, were measured. From days 14 to 16, all treatments significantly reduced tumor volume (P?<?0.05). From days 10 to 16, improvements in the tumor-free body weight were observed in the RGZ and RGZ+imidapril groups. In addition, significant improvements in both gastrocnemius muscle and epididymal adipose mass were observed in all treatment groups (all, P?<?0.05). Furthermore, all treatments significantly increased tumor necrosis factor alpha levels as compared to those observed in the healthy control animals (P?<?0.001). Insulin levels significantly increased in the placebo group as compared to those in the healthy control group (P?<?0.05), which were reduced in all the treatment groups (P?<?0.05). Finally, whereas all treatments significantly reduced atrogin-1 levels as compared to the placebo group (all, P?<?0.05), significant reductions in MuRF-1 levels were only observed in the RGZ and RGZ+imidapril groups (both, P?<?0.05). Thus, all three treatments reduce tumor growth and alleviate cancer cachexia; however, synergistic effects of RGZ+imidapril combination therapy were not observed.     

CA 19-9 as a prognostic index after resection for pancreatic cancer

Serial serum CA 19-9 assays were performed in 30 consecutive patients who underwent resection for pancreatic cancer. Patients with preoperative CA 19-9 levels < 200 U/ml had significantly better prognosis than those with CA 19-9 > 200 Ulml (P < 0.001). Serum tumor marker normalized in 14 patients after tumor resection, and survival in this group was significantly higher than that of patients with persistently elevated CA 19-9 (P < 0.0001). Prognosis was also influenced by absence of lymph node metastases (P < 0.02) and radicality of resection (P < 0.005). Elevation of serum CA 19-9 levels after operation well predicted tumor recurrence from 1-10 months before clinical and radiological evidence. CA 19-9 determination is useful as a prognostic index after resection for pancreatic carcinoma and as a surveillance test in monitoring the efficacy of treatment. © 1993 Wiley-Liss, Inc.     

Prognostic significance of tumor-host interaction in clinical gastric cancer: Relationship between dna ploidy and dendritic cell infiltration

DNA ploidy of tumor cells and the degree of infiltration of dendritic cells were determined in 93 gastric cancer tissue specimens, and the mechanisms of tumor-host interaction on the prognosis were investigated. DNA ploidy patterns were grouped into low and high ploidy, and the degree of infiltration of dendritic cells (DC) was graded into marked and slight infiltration. In the low ploidy group, the 5-year survival rates in patients with marked and slight DC infiltration were 80.7% and 61.5%, respectively (P < 0.05). In the high ploidy group, however, there were no significant differences. In cases of low ploidy, the incidence of lymph node metastasis was significantly lower in the marked DC infiltration group compared with findings in the slight DC group. Thus, markedly infiltrating dendritic cells in gastric cancer tissue may lead to prolongation of survival time for patients with a carcinoma of the low ploidy profile, by preventing widespread nodal involvement. © 1993 Wiley-Liss, Inc.     

Cytomorphologic features of poorly differentiated thyroid carcinoma

BACKGROUND:

Poorly differentiated thyroid carcinoma (PDTC) is an uncommon and aggressive malignancy. Despite the significant clinical implications of a diagnosis of PDTC, its cytomorphologic features have not been well defined. Statistical analysis was applied to a series of 40 PDTCs to identify a specific set of cytomorphologic features that characterized these tumors on fine‐needle aspiration biopsy (FNAB).

METHODS:

In total, 40 thyroid FNABs that were highly diagnosed histologically as PDTC (19 insular carcinomas and 21 noninsular carcinomas) comprised the study group. A control group of 40 well differentiated thyroid neoplasms were selected for comparison. All FNABs were reviewed and scored for a series of 32 cytomorphologic features. The results were evaluated using univariate and stepwise logistic regression (SLR) analyses.

RESULTS:

In univariate analysis, 17 cytomorphologic features were identified that characterized the 40 PDTCs: insular, solid, or trabecular cytoarchitecture (P < .001); high cellularity (P = .007); necrosis (P = .025) or background debris (P = .025); plasmacytoid appearance (P = .0007); single cells (P < .0001); high nuclear/cytoplasmic ratio (P < .0001); scant cytoplasm (P = .03); nuclear atypia (P < .0001), including nuclear pleomorphism (P = .0052) and anisokaryosis (P < .0001); granular/coarse chromatin (P = .026); naked nuclei (P = .01); mitotic activity (P = .0001) and apoptosis (P < .0001); endothelial wrapping (P = .0053); and severe crowding (P < .0001). In logistic regression analysis, severe crowding (P = .0008) and cytoarchitecture (P < .0001) were identified as the most significant cytomorphologic features of PDTCs, and the combination of cytoarchitecture, severe crowding, single cells, and high nuclear/cytoplasmic ratio was the most predictive of PDTC.

CONCLUSIONS:

PDTCs have characteristic cytomorphologic features. By using logistic regression analysis, the features that were identified as the most predictive of PDTC were severe crowding, insular/solid/trabecular morphology, single cells, and high nuclear/cytoplasmic ratio. Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.     

The correlation between IGF-II and Bcl-2 expression in colorectal adenocarcinoma

Our aim for this study was to investigate the correlation and clinical significance between the expression of IGF-II and Bcl-2 in colorectal adenocarcinoma, especially in terms of the metastasis of colorectal adenocarcinoma. Sixty paraffin embedded samples of colorectal adenocarcinoma were selected, and fifteen normal colorectal tissues were used as controls. IGF-II mRNA was detected using in situ hybridization, and the expression of Bcl-2 along with the proliferating cell nuclear antigen (PCNA) protein was detected through immunohistochemistry. The TUNEL assay was used to detect apoptosis. Specimens with a positive cell ratio less than 30% were defined as negative. The levels of IGF-II mRNA and the Bcl-2 protein were significantly higher in colorectal adenocarcinoma (39.64 ± 7.38% and 30.74 ± 7.22%, respectively) than in normal colorectal tissues (22.56 ± 4.21% and 12.17 ± 1.94%, respectively) (P < 0.01). The levels were related to Dukes′ stage and lymph node metastases, but were unrelated to patient age, gender, tumor site, tumor size, and tumor differentiation. Also, a negative correlation was observed between IGF-II mRNA and Bcl-2 protein (P < 0.05) during Dukes′ stages. In addition, a positive correlation between IGF-II mRNA and PCNA or apoptosis, as well as a negative correlation between Bcl-2 and apoptosis were observed (P < 0.01). There was no correlation between Bcl-2 and PCNA (P > 0.05). The patients detected as IGF-II mRNA (+) and Bcl-2 (−) showed the worst prognosis. The expression of IGF-II and Bcl-2 was correlated with the clinical manifestation of colorectal adenocarcinoma; thus, the assessment of both IGF-II and Bcl-2’s status will provide important information regarding the diagnosis and prognosis of colorectal adenocarcinoma.