Non-tumor Group Research Articles

Discrimination of non‐melanoma skin lesions from non‐tumor human skin tissues in vivo using Raman spectroscopy and multivariate statistics

Raman spectroscopy was used to discriminate human non-melanoma skin lesions from non-tumor tissues in vivo. This work proposed the discrimination between non-melanoma (basal cell carcinoma, BCC; squamous cell carcinoma, SCC) and pre-cancerous lesions (actinic keratosis, AK) from benign lesions and normal (non-tumor group, NT) tissues, using near-infrared Raman spectroscopy with a Raman probe. Prior to surgery, the spectra of suspicious lesions were obtained in situ. The spectra of adjacent, clinically normal skin were also obtained. Lesions were resectioned and submitted for histopathology. The Raman spectra were measured using a Raman spectrometer (830 nm). Two types of discrimination models were developed to distinguish the different histopathological groups. The principal components analysis discriminant analysis (PCA/DA) and the partial least squares discriminant analysis (PLS/DA) were based on Euclidean, quadratic and Mahalanobis distances. PCA and PLS spectral vectors showed spectral features of skin constituents, such as lipids (between 1,250 cm(-1) and 1,300 cm(-1) and at 1,450 cm(-1)) and proteins (between 870 cm(-1) and 940 cm(-1), 1,240 cm(-1) and 1,271 cm(-1), and at 1,000 cm(-1) and 1,450 cm(-1)). Despite the small spectral differences between malignant lesions and benign tissues, the algorithms discriminated the spectra of non-melanoma skin and pre-cancerous lesions from benign and normal tissues, with an overall accuracy of 82.8% and 91.9%, respectively. PCA and PLS could discriminate Raman spectra of skin tissues, opening the way for an in vivo optical diagnosis.

ED-16 * HFE POLYMORPHISMS AFFECT SURVIVAL OF BRAIN TUMOR PATIENTS

The HFE protein plays a key role in the regulation of body iron. HFE polymorphisms (H63D and C282Y) are the common genetic variants in Caucasians. Based on frequency data, both HFE polymorphisms have been associated with increased risk in a number of cancers. The prevalence of the two major HFE polymorphisms in a human brain tumor patient population and the impact of HFE polymorphisms on survival have not been studied. In the present study, male GBM patients with H63D HFE polymorphism (H63D) have poorer overall survival than wild type (WT) male GBM (p = 0.03). In GBM patients with the C282Y HFE polymorphism (C282Y), female patients have shorter survival than male patients (p = 0.05). In addition, female metastatic brain tumor patients with C282Y have shorter survival times post diagnosis than WT patients (p = 0.02) or male metastatic brain tumor patients with C282Y (p = 0.02). There is a tendency toward a lower proportion of H63D genotype in GBM patients than a non-tumor control group (p = 0.09) or other subtypes of brain tumors. In conclusion, our study suggests that HFE genotype impacts survival of brain tumor patients in a gender specific manner. We previously reported that glioma and neuroblastoma cell lines with HFE polymorphisms show greater resistance to chemo and radiotherapy. Taken together, these data suggest HFE genotype is an important consideration for evaluating and planning therapeutic strategies in brain tumor patients. [This project was supported in part by the Tara Leah Witmer Endowment and Pennsylvania Department of Health Commonwealth Universal Research Enhancement (CURE) Program.]

Carbohydrate antigens as potential biomarkers for the malignancy in patients with idiopathic deep venous thrombosis: a retrospective cohort study.

A variety of biomarkers have been identified in recent prospective and retrospective reports as being potentially predictive of venous thromboembolis (VTE), particularly idiopathic deep venous thrombosis (IDVT). This study identified a serum tumor biomarker for early screening of IDVT. A total of 128 IDVT patients (54 females and 74 males; average age: 50.9±17.4 years) were included. Carcinoembryonic antigen (CEA), ferritin, β2-microglobulin, cancer antigen (CA) 125, CA 15-3, CA 19-9, squamous cell carcinoma antigen (SCC), alpha-fetoprotein (AFP), prostate specific antigen (PSA), free PSA (f-PSA), and beta-human chorionic gonadotropin (β-HCG) in patients with IDVT were detected. Malignancies were histo- or cytopathologically confirmed. Of the 128 IDVT patients, 16 (12.5%) were found to have malignancies. Serum CEA, CA 125, CA 15-3, and CA 19-9 were found to be helpful for detecting malignancies in IDVT patients. Our study revealed a positive association between these markers and tumors in IDVT patients. On the other hand, SCC and AFP were not sensitive enough to be markers for detecting tumors in patients with IDVT. No significant differences were found in positive rates of ferritin and β2-microglobulin between tumor and non-tumor groups, and no significant difference exists in serum levels of ferritin and β2-microglobulin between the two groups. Carbohydrate antigens, CA 15-3 in particular, may be useful for differential diagnosis and prediction of malignancies in patients with IDVT.

Child-related characteristics predicting subsequent health-related quality of life in 8- to 14-year-old children with and without cerebellar tumors: a prospective longitudinal study.

We identified child-related determinants of health-related quality of life (HRQoL) in children aged 8-14 years who were treated for 2 common types of pediatric brain tumors. Questionnaire measures of HRQoL and psychometric assessments were completed by 110 children on 3 occasions over 24 months. Of these 110, 72 were within 3 years of diagnosis of a cerebellar tumor (37 standard-risk medulloblastoma, 35 low-grade cerebellar astrocytoma), and 38 were in a nontumor group. HRQoL, executive function, health status, and behavioral difficulties were also assessed by parents and teachers as appropriate. Regression modeling was used to relate HRQoL z scores to age, sex, socioeconomic status, and 5 domains of functioning: Cognition, Emotion, Social, Motor and Sensory, and Behavior. HRQoL z scores were significantly lower after astrocytoma than those in the nontumor group and significantly lower again in the medulloblastoma group, both by self-report and by parent-report. In regression modeling, significant child-related predictors of poorer HRQoL z scores by self-report were poorer cognitive and emotional function (both z scores) and greater age (years) at enrollment (B = 0.038, 0.098, 0.136, respectively). By parent-report, poorer cognitive, emotional and motor or sensory function (z score) were predictive of lower subsequent HRQoL of the child (B = 0.043, 0.112, 0.019, respectively), while age at enrollment was not. Early screening of cognitive and emotional function in this age group, which are potentially amenable to change, could identify those at risk of poor HRQoL and provide a rational basis for interventions to improve HRQoL.

Sperm concentration, hyaluronic acid-binding capacity, aneuploidy and persistent histones in testicular cancer.

Are the quantitative and qualitative characteristics of semen samples of patients with testicular cancer (TC), prior to anticancer therapy, different from infertile oligozoospermic (IO) and normozoospermic (NZ) age-matched men? Sperm concentration in TC patients was significantly decreased with no difference in estimated numerical chromosome aberrations and nuclear decondensation compared with NZ men, while the infertile, oligozoospermic men had significantly poorer sperm qualitative characteristics versus the TC group overall and oligozoospermic patients with TC. Spermatogenesis is altered in TC patients at the time of diagnosis. However, the mechanism responsible for the decreased semen quantity in patients with TC is not well understood. Anticancer treatment may have gonadotoxic side effects and post-treatment fertility cannot be predicted. Before commencing anticancer treatment, cryopreservation may be suggested to preserve fertility but there are no data regarding the risk of genetic aberrations in these sperms. This is a cross-sectional study examining semen from 28 patients with TC, 20 IO and 20 NZ age-matched men attending the Andrology Center and the Sperm Cryopreservation Laboratory of the Medical and Health Science Center, University of Debrecen. Semen samples from patients with TC were collected after orchidectomy, but prior to anticancer treatment. Semen samples from TC patients recruited over a period of 4 years were studied. Based on their sperm concentration, TC patients were subgrouped into an oligozoospermic TC (TCO) and a normozoospermic TC group. For statistical analysis, the normal group (NZ + IO) comprised non-tumorous NZ and IO men. The ejaculates were assessed as per World Health Organization guidelines. Hyaluronic acid (HA)-binding capacity was the functional test. To determine the numerical chromosome aberrations, we used multi-color fluorescence in situ hybridization. Aniline blue (AB) staining was performed as a nuclear decondensation marker test. The results did not reveal any significant difference in disomy of sex chromosomes and chromosome 17, diploidy and estimated numerical chromosome aberrations and AB staining results upon comparing the NZ and TC groups, although the sperm concentration (P < 0.001) and HA-binding capacity (P < 0.001) were lower in the TC group. Estimated numerical chromosome aberrations (P < 0.001), AB staining (P < 0.001) and HA-binding capacity (P = 0.019) were lower in the infertile, oligozoospermic group when compared with the patients with TC. The TCO group had significantly better results in every examined parameter than the infertile, oligozoospermic group. In the non-tumorous control group (NZ + IO), a significant (P < 0.001) correlation (Spearman's rho = r) was found between sperm concentration and aneuploidy rate (r = -0.642), AB staining (r = -0.876) and HA binding (r = 0.842); the HA-binding capacity was related to the aneuploidy rate (r = -0.678) and the AB staining (r = -0.811); and there was significant correlation between aneuploidy and AB staining (r = 0.559). In the TC group, apart from the negative correlation between sperm concentration and estimated chromosomal aberrations (r = -0.642), no other correlations were observed. Data on confounders influencing sperm characteristics, such as smoking, occupational or environmental hazards, alcoholism, co-morbidities and other andrological conditions, were not collected. This is the first study to demonstrate that sperm qualitative characteristics in anticancer therapy naïve oligozoospermic TC patients differ significantly from those in IO men and do not differ from those in NZ men. Our results need to be validated in similar groups of men and in other patient groups with cancer where cryopreservation is advisable. This research was supported by the European Union and the State of Hungary, co-financed by the European Social Fund in the framework of TÁMOP-4.2.4.A/2-11/1-2012-0001 'National Excellence Program'. The authors have no conflict of interest to declare.

Expression of osteopontin in central nervous system leukemia and its clinical significance

This study was aimed to investigate the expression of osteopontin (OPN) in central nervous system leukemia (CNSL) and to understand its clinical significance. The expression level of OPN in serum of 62 pediatric patients (22 cases of CNSL, 20 cases of acute leukemia without extramedullary infiltration and 20 cases of nontumor patients) and 19 cases of CNSL with complete remission (CR)were assayed by ELISA; the expression changes of OPN mRNA in bone marrow of the CNSL patients were detected by RT-PCR. The results indicated that the serum OPN level was significantly higher in CNSL group (25.21 ± 6.87 ng/ml) than that in acute leukemia group (13.24 ± 2.73 ng/ml) (P < 0.001) and nontumorous group (3.14 ± 1.60 ng/ml) (P < 0.001); the serum OPN level (4.35 ± 1.50 ng/ml) in CNSL group with CR decreased obviously (P < 0.001) after therapy; RT-PCR analysis showed that the expression of OPN mRNA was higher in CNSL group as compared with other two groups (P < 0.01). It is concluded that the OPN expression may play a role in central nervous system infiltration of leukemia, the mechanism of which remains to need further clinical exploration.

Characteristic power Doppler sonographic images of tumorous and non-tumorous buccal space lesions

The aim of this study was to evaluate the characteristic power Doppler sonographic images of buccal space tumorous and non-tumorous lesions. 48 patients with buccal space lesions were evaluated with greyscale sonography followed by power Doppler sonography with a 12 MHz linear transducer. On greyscale sonography, buccal space lesions were assessed for the boundary (clear or unclear), echogenicity (hypoechoic or isoechoic) and internal architecture (homogeneous or heterogeneous). Power Doppler sonography was performed to evaluate the vascular signals within the buccal space lesions. 48 lesions were found in the 48 patients; of these 48 lesions, 28 were tumourous and 20 were non-tumourous. In the 28 tumours, 15 cases showed clear boundaries, 15 cases were hypoechoic relative to adjacent tissues and 22 cases presented with a heterogeneous appearance on greyscale sonography. The internal vascularity of 19 tumours was shown using power Doppler sonography. In the 20 non-tumorous lesions, 11 cases showed clear boundaries, 17 cases were hypoechoic relative to adjacent tissues and 13 cases presented with a homogeneous appearance on greyscale sonography. 18 non-tumorous lesions showed no internal vascularity using power Doppler sonography. Logistic multivariate regression analysis between the tumour group and the non-tumorous lesions group demonstrated that the internal architecture (odds ratio = 8.270, p = 0.029) and vascular signals (odds ratio = 17.533, p = 0.003) were significant variables. Power Doppler sonography is a useful technique for the differential diagnosis of tumorous and non-tumorous buccal space lesions.

Circulating miR-125b is a novel biomarker for screening non-small-cell lung cancer and predicts poor prognosis

MicroRNAs are small, non-coding RNAs that are critical regulators of various diseases including cancer, and may represent a novel class of cancer biomarkers. Recent reports have highlighted the oncogenic aspects of miR-125b. However, the level and clinical relevance of circulating miR-125b transcripts in human serum of non-small-cell lung cancer (NSCLC) patients are unclear. The purpose of this study was to identify circulating miR-125b transcripts in human serum for use as a biomarker for stratification and prediction of prognosis in NSCLC. We analyzed serum levels of miR-125b in 193 patients with different stages of NSCLC. Blood samples were collected before surgery and therapy. Quantitative reverse transcription-polymerase chain reaction of circulating miR-125b transcripts was performed directly in serum to improve the efficiency of miRNA assessment. Receiver operating characteristic analysis was used to evaluate the sensitivity and specificity of serum miR-125b. We found that serum miR-125b was consistently expressed in the non-tumor group and was significantly associated with NSCLC stage. miR-125b expression was capable of separating NSCLC patients from control groups with an area under the curve of 0.786. Furthermore, patients with high miR-125b expression displayed a significantly poorer prognosis compared with patients with low expression (p<0.0001). Multivariate analysis indicated that high miR-125b expression was an independent prognostic factor for survival. We propose that serum miR-125b may represent a novel biomarker in NSCLC patients and that high miR-125b expression is an independent prognostic factor for survival.

The Effects of Electroacupuncture at the ST36 (Zusanli) Acupoint on Cancer Pain and Transient Receptor Potential Vanilloid Subfamily 1 Expression in Walker 256 Tumor-Bearing Rats

Several studies have addressed the expression of transient receptor potential vanilloid subfamily 1(TRPV1) playing an important role in the generation of cancer pain. Electroacupuncture (EA) is an effective method of acupuncture shown to attenuate different kinds of pain such as inflammatory, neuropathic, and cancer. In this study, we investigated the effect of EA on cancer pain caused by intraplantar injection of Walker 256 carcinoma cells and cancer-driven TRPV1 expression in the dorsal root ganglions (DRGs). Rats were randomly divided into 4 groups: the nontumor cell inoculation group (normal control, n = 8); Walker 256 carcinoma cell inoculation group (tumor control, n = 8); sham point electrical stimulation treatment with Walker 256 carcinoma cell inoculation group (SES, n = 8); EA treatment with Walker 256 carcinoma cell inoculation group (EA, n = 8). The time courses of thermal, mechanical sensitivity, and spontaneous nocifensive behavior were determined. In addition, TRPV1 expression in DRGs was observed by quantitative real-time polymerase chain reaction and Western blotting. Injection of cancer cells decreased the paw withdrawal threshold, increased spontaneous nocifensive behavior, and induced significant thermal hyperalgesia that was attenuated by EA at the ST36 acupoint (2 Hz, 0.3 ms, ≤1 mA). TRPV1 mRNA and protein in DRGs were upregulated in the cancer pain model, and EA at ST36 acupoint counteracted the cancer-driven upregulation of TRPV1 expression in the corresponding DRGs. EA at ST36 could attenuate cancer-induced pain, at least in part, through suppressing TRPV1 mRNA and protein upregulation in the DRGs.

Expression of MCP1 gene in serum and tumor tissues in bladder urothelial carcinoma patients and its clinical significance

Objective To investigate the monocyte chemoattractant protein (MCP1) gene expression of the bladder urothelial carcinoma and its correlation with the pathogenesis of the bladder urothelial carcinoma.Methods Thirty cases of patients with the bladder urothelial carcinoma, including 20 cases of male and 10cases of female, were taken the blood and bladder tissue.In control group, 30 cases of non-cancer patients,including 20 cases of male and 10 cases of female, were taken the blood samples.ELISA method was used to detected the concentration of plasma MCP1, immunohistochemical method to investigate the expression of MCP1 in the bladder urothelial carcinoma and adjacent tissues.Real-time quantitative RT-PCR was applied to detected the expression of MCP1. Data of the two groups were comparied and the relationship between the expression of MCP1 and the clinical characteristics of the bladder urothelial carcinoma was analyzed.Results MCP1 in group of patients with the bladder urothelial carcinoma was (193.4±105.7) pg/ml, and higher than that in non-tumor group (91.8±34.6) pg/ml (t = 8.37, P ＜0.001).MCP1 in invasive bladder cancer was (204.3±167.5) pg/ml and superficial bladder cancer was (130.6±69.2) pg/ml (t = 2.667, P = 0.013). By immunohistochemistry, the MCP-1 positive rate in the bladder urothelial carcinoma was 70.0 % (21/30), that in adjacent cancer tissue was 43.3 % (13/30) (χ2 = 4.9, P ＜0.05). The positive rate of MCP1 in invasive bladder cancer in tumor group was 80.0 % (8/10) and that in superficial bladder cancer was 65.0 %(13/20).At the same time, MCP- 1 positive intensity in the bladder urothelial carcinoma was significantly higher than that in adjacent tissues. The intensity in invasive bladder cancer was higher than that in superficial ones. Total RNA and mRNA levels of MCP-1 in the bladder urothelial carcinoma were statistically differences compared with that in adjacent tissues (χ2 = 10.08, P ＜0.05).Conclusion The upregulation of MCP1 gene expression is likely to play an important role in the incidence and metastasis of the bladder urothelial carcinoma. Key words: Urinary bladder neoplasms; Chemokine CCL2; Gene

Simultaneous detection of expression and gene mutations of HER2/neu in Chinese patients with gastric cancer

HER2/neu is one of the few identified oncogenes in tumorigenesis. Attention has been focused on the potential effect of HER2/neu mutations in the tyrosine kinase domain on carcinoma-targeted therapy. However, data concerning HER2/neu mutations in Chinese patients with gastric cancer (GC) are limited. This study aimed to detect the expression and somatic mutations of HER2/neu in Chinese patients with GC. Immunohistochemical staining for HER2/neu was performed on 72 formalin-fixed, paraffin-embedded specimens of GC (40 intestinal and 32 diffuse type). The correlation between the overexpression of HER2/neu and clinicopathological parameters was statistically analyzed. Somatic mutations in the tyrosine kinase domain of HER2/neu in the 72 patients were detected by direct sequencing. In the GC group, overexpression of HER2/neu was detected in 13 of the 72 GC patients and in 4 of the 72 adjacent tissues in the non-tumorous group (18.1 vs. 5.6%, P<0.05). Furthermore, the intestinal type of GC exhibited a higher rate of HER2/neu overexpression than the diffuse type (29.7 vs. 5.7%, P<0.05). The rate of HER2/neu overexpression in stage III-IV (TNM stage) GC cases was significantly higher than that in stage I-II (28.2 vs. 6.6%, P<0.05). HER2/neu overexpression correlated with a significantly less favorable patient survival (P=0.046). No somatic mutations in the tyrosine kinase domain of HER2/neu were detected in tumor tissues or the corresponding non-tumorous ones in the specimens obtained from the 72 Chinese GC patients. Results suggest that overexpression of HER2/neu is a frequent molecular event strongly associated with a poor patient prognosis, whereas the incidence of somatic mutations of the HER2/neu kinase domain is more likely a low-frequency event in Chinese GC patients.

Comparative Study on Overexpression of HER2/neu and HER3 in Gastric Cancer

Owing to the special importance of the HER family in tumorigenesis, the downstream signaling pathways and effectors have become the key molecules in the strategy of carcinoma-targeted therapy. Recent evidence that HER3 is responsible for tumor resistance to therapeutic agents targeting EGFR or HER2/neu, along with the new findings that HER3 is involved in the process of dedifferentiation of gastric cancer (GC) have highlighted the critical role of HER3 in cancer research. HER3 is becoming a new targeted molecule in cancer treatment. Here, we comparatively investigated the expression of HER2/neu and HER3 in gastric cancer of two pathologic types (intestinal type and diffuse type) using immunohistochemistry (IHC) and analyzed the correlation between overexpression of HER2 and HER3 and clinicopathologic parameters. An IHC study for HER2 and HER3 was performed on 102 formalin-fixed, paraffin-embedded specimens of GC-60 intestinal and 42 diffuse types. The correlation between overexpression of HER2 and HER3 and clinicopathologic parameters was statistically analyzed. In the GC group, overexpression of HER2 and HER3 was detected in 19 (18.6%) and 14 (13.7%) of 102 GC patients, respectively. In a nontumorous group of 102 specimens, 5 were HER2-positive (4.9%) (18.6% vs. 4.9%, p < 0.01), and 2 were HER3-positive (2.0%) (13.7% vs. 2.0%, p < 0.01). No co-overexpression of HER2 and HER3 was observed. The intestinal type of GC exhibited a higher rate of HER2 overexpression than did the diffuse type (26.7% vs. 7.1%, p < 0.05), whereas the diffuse type of GC exhibited a significantly higher rate of HER3 overexpression than did the intestinal type (26.2% vs. 5.0%, p < 0.01). The overexpression rates of HER2 and HER3 in phase III-IV (TNM stage) disease were significantly higher than that in phase I-II disease (24.0% vs. 7.7%, p < 0.05 and 22.0% vs. 5.8%, p < 0.05, respectively). HER2 and HER3 overexpression was also correlated with a significantly worse survival (p = 0.046 and 0.024, respectively). The selective overexpression of HER2 and HER3 in the two histologic types of gastric cancer is strongly associated with a poor prognosis. Being an important member of the HER family, HER3 may become another candidate for molecular-targeted therapy in gastric cancer, especially for the diffuse histologic type.

Radio-iron uptake of rat hepatoma.

Iron uptake rate of the tumor tissues of the rat liver was compared with that of the nontumor tissues of the same rat liver. Ferric[59Fe] citrate was injected to rats bearing hepatoma induced by the administration of diethylnitrosamine. The tumor and nontumor tissues were enucleated from the liver and radioactivity (cpm/10 mg tissue) of the individual tissue was calculated to estimate the iron uptake rate of the tissue. Eleven rats were finally available for the study. Population mean of radioactivity of the tumor group was larger than that of the nontumor group when examined 6 hr after the injection, so far as the specimens of both groups were sufficient in number, more than 19. The variance of radioactivity of the tumor group was always larger than that of the nontumor group, probably in part due to cytological and histological varieties of the tumor tissues. These results suggest that the tumor tissues can take up more iron than the nontumor tissues.

β-Hydroxy-β-methylbutyrate supplementation reduces tumor growth and tumor cell proliferation ex vivo and prevents cachexia in Walker 256 tumor-bearing rats by modifying nuclear factor-κB expression

Cancer cachexia syndrome contributes to wasting and weight loss leading to inefficacy of anticancer therapy. In this study, the anticatabolic agent beta-hydroxy-beta-methylbutyrate (HMB) was supplemented to adult Walker 256 tumor-bearing rats during 8 weeks aiming to determine if tumor burden could be reduced. Male Wistar rats were randomly assigned to nontumor and tumor-bearing groups and fed regular chow or regular chow plus HMB supplemented (76 mg/kg body weight). Beta-hydroxy-beta-methylbutyrate supplementation induced a lower tumor weight and tumor cell proliferation ex vivo, totally prevented glycemia reduction, as well as blunted the increase in the serum lactate concentrations and also preserved glycogen stores in tumor-bearing rats. Reduction in tumor cell proliferation ex vivo was accompanied by increased nuclear factor-kappaB inhibitor-alpha content by more than 100%. In contrast, nuclear factor-kappaB p65 subunit content was suppressed by 17% with HMB supplementation. In conclusion, HMB supplementation, at a similar dose used in humans to increase muscle mass, caused antitumor and anticachectic effects, with tumor-cell nuclear factor-kappaB pathway participation, which might be a potential nutritional strategy in cancer therapy.

Relationship between biomarker expression and allelic alteration in esophageal carcinoma

Expression of biomarkers and probable allelic alterations were studied in esophagus tissue samples from patients with esophageal carcinoma. A total of 116 esophagus tissue samples were obtained from 25 patients with esophagus cancer. Histological studies revealed 23 samples were adenocarcinoma and 14 samples were epidermoid carcinoma while 79 samples were non-tumor. Expression of biomarkers was determined by enzyme immunoassay, and allelic alterations on chromosome 17p were performed by polymerase chain reaction (PCR) using primers D17S513 and D17S514. The adenocarcinoma group exhibited an increase of matrix metalloproteinase (MMP)-1 (P < 0.0001) and sialyl Le (a) (P < 0.001) mean levels when compared with the non-tumor group. Adenocarcinoma samples from patients with more than three positive lymph nodes had lower levels of tissue-inhibitor metalloproteinase (TIMP)-1 than those with negative nodes (P < 0.0005). Positive allelic alteration was associated with high levels of MMP-1 expression (P = 0.003). Epidermoid carcinoma samples showed higher expression of MMP-1 (P < 0.0001) and TIMP-1 (P < 0.02) than non-tumor samples. Both epidermal growth factor receptor and sialyl Le (a) levels were overexpressed in tumors of patients with more than three positive lymph nodes (P < 0.005). Carcinoembryonic antigen levels were higher in tumors associated with allelic wild type group (P = 0.0001) and patients with negative lymph nodes (P < 0.05). Furthermore, variability in expression of biomarkers was observed according to sample location, and allelic alterations were also found both in tumor and in some non-tumor samples. The data suggest that overexpression of tissue biomarkers associated with allelic alterations may have potential prognostic implications with different behavior in esophagus cancer.

The correlation between the dilated extent of bile duct and gallbladder and low bile duct obstructive jaundice diseases

目的 探讨肝内、外胆管及胆囊扩张程度、形式对低位梗阻性黄疸的诊断价值.方法回顾性分析105例低位阻塞性黄疸患者的CT和内窥镜逆行胰胆管造影(ERCP) 的影像资料,将胆系扩张分为下列7种类型:Ⅰ型为肝内、外胆管及胆囊均重度扩张;Ⅱ型为肝外胆管、胆囊重度扩张伴肝内胆管轻度扩张;Ⅲ型为肝内、外胆管重度扩张伴胆囊不扩张或轻、中度扩张;Ⅳ型为肝外胆管重度扩张伴肝内胆管和胆囊不扩张或轻、中度扩张;Ⅴ型为肝内胆管重度扩张伴肝外胆管及胆囊不扩张或轻、中度扩张;Ⅵ型胆囊重度扩张伴肝内、外胆管不扩张或轻、中度扩张;Ⅶ型为肝内、外胆管及胆囊均不扩张或轻、中度扩张.结合手术和病理结果,分析各类型胆系扩展和其低位阻塞性黄疸病变的相关性.结果 105例低位阻塞性黄疸中33例为肿瘤性病变,72例为非肿瘤性病变.肿瘤性病变中,Ⅰ型16例,Ⅱ型10例,Ⅲ型4例,Ⅳ型1例,Ⅶ型2例.非肿瘤性病变中,Ⅰ型4例,Ⅱ型4例,Ⅲ型9例,Ⅳ型33例,Ⅴ型2例,Ⅵ型11例,Ⅶ型9例.Ⅰ、Ⅱ型扩张和Ⅲ～Ⅶ型扩张在肿瘤和非肿瘤病变中差异有统计学意义(χ2=47.33,P<0.01).结论低位阻塞性黄疸病变性质和肝内、外胆管及胆囊扩张程度密切相关:(1)Ⅰ型和Ⅱ型扩张提示绝大多数为肿瘤性病变,少数为嵌顿性结石.(2)Ⅲ～Ⅶ型扩张常为胆管、胆囊结石及炎症。

Production of Thymus and Activation-Regulated Chemokine and Macrophage-Derived Chemokine by CCR4+ Adult T-Cell Leukemia Cells

Adult T-cell leukemia/lymphoma (ATL) is a peripheral CD4(+)CD25(+) T-cell malignancy caused by human T-cell leukemia virus type I. The tumor cells frequently infiltrate in the skin, lymph nodes and other organs and especially form prominent cutaneous masses. Recently, ATL cells have been shown to express Th2 chemokine receptor CCR4. The aim of this study is to investigate the possibility that CCR4 ligands, thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC), are produced by CCR4(+) ATL cells per se. CD4(+) or CD4(+)CD14(-) cells were purified from peripheral blood mononuclear cells of 11 ATL patients with cutaneous involvement and normal healthy volunteers. Tissue-infiltrating cells were isolated from skin tumors. The expression of chemokine receptors on these cells were analyzed by flow cytometry. The production of chemokines and cytokines by the neoplastic cells was assessed by ELISA and reverse transcription-PCR after cultivation for 96 hours in the presence or absence of anti-CD3/CD28 monoclonal antibodies. Finally, TARC and CCR4 expressions were examined by immunohistochemistry. ATL cells highly expressed CCR4 but did not necessarily exhibit the Th2 cytokine profile. The cells also produced TARC and MDC. The production level of MDC was higher in the skin tumor formation group than that in the nontumor group. Immunohistochemically, both CCR4 and TARC were expressed by the tumor cells in the lesional skin. ATL cells not only express CCR4 but also produce TARC and MDC. The skin tumor formation as well as the monoclonal integration of proviral DNA are the factors that are associated with the high production of Th2 chemokines by ATL cells.

Life span and spontaneous tumors incidence of the Wistar Mishima (WM/MsNrs) rat.

The life spans and spontaneous tumors in a total of 1960 Wistar Mishima (WM/MsNrs) rats, inbred strain, from the 80-130th generations were examined. The average life span (mean +/- SD) was 731 +/- 173 days (n = 1053) in the males and 813 +/- 214 days (n = 907) in the females (p < 0.0001). The average life span of tumor-afflicted females was significantly longer than that of the non-tumor group (p < 0.0001), while no such difference was observed in males. Tumors were observed in 33 males (3.1%) and 246 females (27.1%). In the males, tumors were often observed under the skin (2.2%). Frequencies of tumors in lung and liver, bones and intestine were less than 0.5%. In the females, incidence of mammary tumor was 20.1%, and various organs such as ovaries, uterus, bones, lung, and liver had tumor incidence frequencies of less than 3.5%. It was concluded that WM/MsNrs rats might be suitable for life span and age-related studies because of their characteristics of length of longevity and the low incidence of spontaneous tumors in both sexes.

Tiny staining spots in liver cirrhosis associated with HCV infection observed by computed tomographic hepatic arteriography: follow-up study.

It is important to distinguish small lesions with increased arterial perfusion observed by computed tomographic arteriography (CT-A) from hepatocellular carcinoma (HCC). However, the clinical characteristics and prognosis of such lesions have not been clarified. We retrospectively examined 200 patients with cirrhosis related to hepatitis C virus (HCV) infection who had undergone both CT-A and CT arterioportography between 1995 and 1998, and found 80 tiny staining spots (TSS)s, with a diameter of 5-10 mm, by CT-A (35 patients). The mean TSS observation period was 29.0 months. If the major axis was larger than 10 mm and showed a 1.5-fold or more increase, the lesion was regarded as tumor growth (TG). The TSS lesions were divided into two groups according to whether the patient had or did not have HCC. The prognosis of TSS was classified into three groups; HCC-suspected group, nontumor group, and unclassified group, in which TG was negative although transcatheter arterial embolization (TAE) had been performed. Of the 40 TSSs in 14 patients without HCC, 2 (5%) were suspected as HCC. Of the 40 TSSs in 21 patients with HCC, 13 (32.5%) were suspected as HCC. There were no significant differences in the size, position, and morphology of TSSs among the three prognostic groups. Of the 7 TSSs with a high signal intensity on T2-weighted magnetic resonance (MR) images, 5 were in the HCC-suspected group. We recommend early treatment of TSSs accompanying HCC or showing features of malignancy at the imaging workup.

Glucose-6-phosphatase gene mutations in 20 adult Japanese patients with glycogen storage disease type 1a with reference to hepatic tumors.

A few cases are reported of liver neoplasms observed in patients with glycogen storage disease type 1a (GSD1a). Genetic analysis was carried out in adult Japanese patients with GSD1a and their family members, and hepatic tumors were also investigated in these patients. DNA was extracted from the peripheral blood lymphocytes of 20 adult patients with GSD1a and 21 family members, and mutations were detected based on the differences in the polymerase chain reaction (PCR) products of the glucose-6-phosphatase (G6Pase) gene shown by single-strand conformation polymorphism (SSCP) analysis. Actual mutations were confirmed by direct sequencing. The relationship between the occurrence of liver tumors and the clinical characteristics of the patients was also investigated. Nineteen of the 20 patients were homozygous for the G727T mutation and one was a compound heterozygote for G727T plus G327A mutations. All of the 19 homozygotes for G727T had hepatomegaly, three had hepatocellular carcinoma, one had cholangiocellular carcinoma, and seven had hepatic adenoma. There were no differences between the tumor and non-tumor groups with respect to laboratory biochemical data (P > 0.05). The mean age of G727T homozygotes with hepatocellular carcinoma was 48.3 years, and that of those with hepatic adenoma was approximately 20 years younger. The G727T mutation seems to be common among Japanese patients with GSD1a, and the discovery of one heterozygote with a combination of G727T and G327A mutations (the latter mutation is common among Chinese) by the use of polymerase chain reaction-single strand conformation polymorphism analysis gave further insight into Japanese ancestry. This is the first study of liver tumors in a large group of adult GSD1a patients with the G727T mutation. As most of the patients in our series are free from other chronic liver diseases such as viral hepatitis, other genetic and/or acquired factors may have influence on the sequel to this metabolic disease.