HO-1在原发性肝癌中的表达及意义

目的探讨血红素氧合酶-1(HO-1)在肝癌组织中的表达及意义。方法分别用RT-PCR、免疫组化检测35例肝癌标本、癌旁组织、正常肝组织的HO-1mRNA及HO-1蛋白表达情况,探讨HO-1在肝癌组织中表达及意义。结果肝癌组织中的HO-1mRNA表达明显高于癌旁组织、正常肝组织(P<0.05),肝癌组织中的HO-1表达高于癌旁组织及正常肝组织(P<0.05)。结论HO-1mRNA、蛋白表达在原发性肝癌组织中表达上调与肝癌演变、发展密切相关,并提示HO-1对肝癌细胞存在保护作用。     

Giant renal oncocytoma masquerading as infiltrating renal cell carcinoma: case report and literature review

Renal oncocytomas (RO) are tumours containing a population of cells with highly differentiated eosinophilic granular cytoplasm, extremely rich in mitochondria. It is estimated they account for about 3 to 7% of all solid renocortical tumours that were previously regarded as renal cell carcinoma. Based on their clinical behaviour and distinct pathologic features they are now regarded as benign renal tumours, often less than 5cm in diameter. We present a case of giant renal oncocytoma in a patient with synchronous bladder tumour, with pre-operative clinical, urographic and ultrasound features of locally advanced renal cell carcinoma (RCC). Findings at surgery included huge right renal tumour with infiltration to the duodenum; hepatic colic flexure; gall bladder; liver capsule and the greater omentum with small indurations at the base of the bladder. There was no tumour extension to the renal vein, no peritoneal seedling, no nodal metastasis and no ascites. Radical nephrectomy was carried out with good prognosis and without recurrence 52months post nephrectomy. This presentation, besides highlighting the possible giant nature of RO, also illustrates the malignant potential of RO to infiltrate contiguous structures and mimics infiltrating RCC. In view of the difficulties at establishing pre-operative diagnosis in this disease and because nephron-sparing surgery is curative, especially for the well-circumscribed tumours, RO should be considered in the management of patients with features of infiltrating RCC. A review of literature is also presented.     

Quantitative changes in glycosaminoglycans in the lungs of rats exposed to diesel exhaust.

Exposure to diesel exhaust (DE) induces lesions in lung epithelium by generation of reactive oxygen species. Glycosaminoglycans (GAG), components of extracellar matrix, are thought to play important roles in cell proliferation and differentiation in the repair process of injured tissue. We investigated how GAG are related to the recovery of lung tissue from injury. Using high-performance liquid chromatography analysis, we determined the amounts of GAG, such as chondroitin sulfate (CS), dermatan sulfate (DS), and hyaluronan (HA) in the lungs of rats exposed to DE for 4 weeks at concentrations of 0.3 or 3 mg/m(3) as suspended particulate matter, or to filtered air. The contents of CS and HA in the surroundings of the bronchi were significantly increased after exposure to DE. In addition, immunohistochemical staining showed that the number of 8-hydroxydeoxyguanosine-positive cells as a marker of cell damage, and proliferating cell nuclear antigen-positive cells also increased in the same areas in which the levels of GAG were elevated in the lungs of rats exposed to 3 mg/m(3) DE. These results suggest that CS and HA in the lung contribute to cell proliferation and remodeling in the process of recovery from injury caused by exposure to DE.     

Role of chondroitin sulfate C in the action of anthrax toxin

Anthrax toxin is produced by Bacillus anthracis, the causative agent of anthrax, and is responsible for the majority of disease symptoms. The toxin consists of 3 proteins, protective antigen (PA), lethal factor (LF), and edema factor (EF), which combine to form lethal and edema toxin. Glycosaminoglycans, which are present on the surface of cells, were investigated with regard to their role in toxicity resulting from anthrax toxin exposure. Lethal toxin-induced cytotoxicity of the RAW 264.7 cells was significantly inhibited by the addition of chondroitin sulfate C as determined by the MTT assay. By contrast, several other glycosaminoglycans, including heparin, heparan sulfate, and dermatan sulfate did not show significant levels of inhibition. Studies utilizing fluorescence-labeled PA demonstrated decreased PA binding to RAW 264.7 cells with the addition of chondroitin sulfate C. Formation of PA oligomers at the surface of cells after binding was also inhibited by chondroitin sulfate C. Interestingly, enzymatic degradation of endogenous chondroitin sulfate C from the cell surface with chondroitinase ABC was accompanied by increased sensitivity to the toxin. These findings were further confirmed by pretreating cells with sodium chlorate to reduce the degree of cell surface glycosaminoglycans sulfation. In addition, chondroitin sulfate C effectively inhibits edema toxin-induced cAMP accumulation in cells. Our results indicate that chondroitin sulfate C may play an important role in the toxicity of anthrax toxin.     

Proteoglycans synthesized by cultured bovine aortic smooth muscle cells after exposure to lead: lead selectively inhibits the synthesis of versican, a large chondroitin sulfate proteoglycan

To investigate the effects of lead on the formation of extracellular matrix in the vascular wall, we characterized proteoglycans synthesized by cultured vascular smooth muscle cells after exposure to the metal by biochemical techniques. Confluent cultures of bovine aortic smooth muscle cells were metabolically labeled with [(35)S]sulfate or [(35)S]methionine/cysteine in the presence of lead nitrate. The amount of glycosaminoglycans (GAGs) was evaluated by the incorporation of [(35)S]sulfate into GAGs by the cetylpyridinium chloride precipitation method. The labeled proteoglycans were characterized by DEAE-Sephacel ion exchange chromatography and Sepharose CL-2B molecular sieve chromatography. The GAG M(r) and composition were analyzed by Sepharose CL-6B chromatography, and the core protein M(r) was analyzed by SDS-polyacrylamide gel electrophoresis, before and after digestion with papain or chondroitin ABC lyase. Lead significantly decreased the [(35)S]sulfate incorporation into GAGs accumulated in the cell layer and the conditioned medium. [(35)S]Sulfate-labeled proteoglycans obtained from the cell layer and the conditioned medium were separated into three peaks on DEAE-Sephacel chromatography and only the peak with the highest charge density was decreased by lead. The highly charged peak was eluted near the void volume on Sepharose CL-2B molecular sieve chromatography and sensitive to chondroitin ABC lyase on Sepharose CL-6B chromatography, indicating that lead selectively inhibits the synthesis of large and highly charged chondroitin/dermatan sulfate proteoglycans (CS/DSPGs). However, the size of chondroitin/dermatan sulfate chains of the CS/DSPGs was M(r) approximately 47000 in both the control and lead-treated cultures. On the other hand, lead decreased the accumulation of a large CS/DSPG with a core protein of approximately 450 kDa in the cell layer and the conditioned medium; the core protein was identified as versican core by Western blot analysis. It is therefore suggested that lead inhibits the synthesis of the versican core protein in vascular smooth muscle cells without a change in length of chondroitin/dermatan sulfate side chains. As a result, versican-poor extracellular matrix would be induced by lead in vascular smooth muscle cells.     

MMP-26与PDZK1在肾透明细胞癌中的表达

目的:研究基质金属蛋白酶-26和PDZK1在不同分化程度肾透明细胞癌中表达,探讨其与临床发生、发展及预后的意义。方法:通过免疫组织化学(ABC法)对60例不同分化程度肾透明细胞癌组织切片(A组)和20例肾脏良性病变的正常组织切片(B组)检测MMP-26和PDZK1阳性细胞的表达情况。结果:MMP-26在A组的阳性表达率为80%,在B组阳性表达率为50%(P<0.05);PDZK1阳性率A组表达为85.0%,在B组表达为30%(P<0.05)。结论:MMP-26和PDZK1在肾透明细胞癌组织中高表达。     

原发性肝癌患者血清血管内皮生长因子检测及临床意义

目的探讨血清血管内皮生长因子（VEGF）与原发性肝癌（PHC）发生、发展、转移、疗效及预后的关系。方法采用ELISA法检测了50例PHC、13例良性肝肿瘤患者和30例正常对照组血清VEGF水平。结果PHC患者血清VEGF水平明显高于正常人及良性肝肿瘤患者（P〈0．01），并与肿瘤直径、AFP水平、有无腹腔积液、有无肝外转移、临床分期及治疗效果有关。结论PHC患者血清VEGF水平在一定程度上可反映肝癌发生、发展、转移情况及治疗效果．可能成为预测肝癌转移及疗效的有效指标。     

A pilot study of nuclear instability in archived renal and upper urinary tract tumours with putative ochratoxin aetiology

DNA ploidy measurement has been applied uniquely to wax-embedded tissue of primary renal cell and metastatic tumours of a key experimental researcher on porcine ochratoxicosis, a control, and four transitional cell carcinomas from cases of Balkan endemic nephropathy. Primary renal tumour was diploid, and hyperdiploid metastasis was within the lower ploidy range for typical renal cell carcinoma. Three Balkan primary tumours showed extensive aneuploidy indicating marked nuclear instability, similar to model rat renal carcinoma caused by ochratoxin A. In contrast, much less nuclear instability in the putative occupational ochratoxicosis case fitted poorly with the ochratoxin A model.     

良恶性甲状腺肿瘤组织中层粘连蛋白的表达及意义

目的比较层粘连蛋白（LN）在良恶性甲状腺肿瘤的表达及其意义。方法采用免疫组化方法检测12例甲状腺正常组织和38例甲状腺腺瘤、20例甲状腺癌组织中LN的表达，对LN表达率作两两比较。结果LN在甲状腺腺瘤与甲状腺癌组织中阳性表达率差异有显著意义（P〈0．05）；在甲状腺癌组织中与甲状腺正常组织中的阳性表达率差异有非常显著意义（P〈0．01）。LN在甲状腺癌细胞及出现于甲状腺滤泡性腺瘤组织中的乳头状腺瘤细胞胞浆中有表达。结论LN参与了肿瘤的发生发展，可作为甲状腺良恶性肿瘤的鉴别诊断指标之一。LN免疫反应物出现于肿瘤细胞胞浆很可能是良性肿瘤转化为癌的标志。     

Isolation of proteoglycans synthesized by rat heart: evidence for the presence of several distinct forms.

1. The proteoglycans (Ps) synthesized by auricle and ventricle from adult rat heart were studied. 2. Auricle tissue incorporated over two times radioactive sulfate compared to ventricle tissue and the Ps were mainly found in the detergent insoluble fraction. 3. The Ps from both tissues were isolated by ion-exchange chromatography on DEAE-Sephacel, followed by gel filtration on Sepharose CL-6B and SDS-PAGE electrophoresis. 4. Enzymatic and chemical degradation of these Ps suggest that at least three and probably four different species of Ps can be observed in heart tissue. 4. A high molecular weight chondroitin sulfate-P, a high molecular weight heparan sulfate-P, a chondroitin/dermatan sulfate-P of 240-200 kDa and a dermatan sulfate of 115 kDa. 5. This latter P was specifically immunoprecipitated using rat decorin antiserum.     

肾脏不典型良性肿块与肾癌的多层螺旋CT鉴别诊断

目的 提高肾脏良恶性肿块的诊断与鉴别诊断水平,降低误诊率,避免误切肾脏.方法 肾良性肿块患者9例,男6例,女3例,年龄30～76岁,平均56岁.肾癌患者52例,男性40例,女性12例,其中透明细胞癌37例,乳头状癌10例以及嫌色细胞癌5例.术前均行CT平扫及三期增强扫描,比较肾脏良性肿块与肾癌的CT影像学特点.结果 所有肾肿块性病变均经手术及病理证实.9例患者中8例按肾癌行根治术,1例行肿瘤剜除术,术后病理均为肾脏良性病变.其中少脂肪血管平滑肌脂肪(AML)4例、嗜酸细胞瘤2例、平滑肌瘤、炎性假瘤和囊肿伴血肿机化各1例.52例肾癌平扫呈均匀或不均匀的等、稍低、稍高或混杂密度;增强扫描肿瘤呈不均匀、均匀明显或不明显强化.结论 CT是肾脏良恶性肿块诊断与鉴别诊断中一种重要的影像学手段,对于影像学表现不典型良性肿块病变者,术前穿刺病理活检或术中冰冻病理切片是避免误诊及误切肾脏的关键,从而选择合适的治疗方案,避免不必要的肾根治性术.     

Treatment of advanced malignancies with ifosfamide under protection with mesna

Fifty-seven patients with advanced malignant tumours were treated with ifosfamide (Holoxan) and mesna (Uromitexan) in our department from November 1979 to December 1984. This series comprised eight cases of soft tissue sarcoma, nine cases of ovarian carcinoma, five cases of non-seminomatous testicular tumour, 11 cases of bronchogenic carcinoma, three cases of renal carcinoma, seven cases of non-Hodgkin's lymphoma, two cases of skeletal fibrosarcoma, two cases of breast carcinoma, one case each of Ewing's tumour, prostatic carcinoma, seminoma, plasma cell tumour, multiple myeloma, malignant teratoma, nasopharyngeal carcinoma, Wilms's tumour, neuroblastoma and mycosis fungoides. Out of these 57 cases, 53 were evaluable. There were five complete remissions and 20 partial remissions, corresponding to a total response rate of 47%. The overall median survival time (MST) of the 53 evaluable patients was 7.5 months. The responders had a longer survival time (MST 10 months) than the non-responders (MST 4.75 months) (p greater than 0.05). Analysis of the results according to sex, age, dosage of ifosfamide and degree of histological differentiation of the tumour cells failed to show any influence of these factors on the therapeutic results. The response rate to ifosfamide found in this study might be related to the histological origin of the tumours and to whether the primary tumours had been resected. The non-seminomatous testicular tumours, non-Hodgkin's lymphomas and ovarian carcinomas showed a high response rate. The response rate was higher in the group in which the primary tumour had been resected (61%) than in the non-resected group (12%) (except the non-Hodgkin's lymphoma). The side-effects of this regimen were moderate. Dyspepsia, nausea, vomiting, myelodepression, dizziness, and alopecia were common. Cystitis could be prevented nearly completely by concomitant administration of mesna, when given correctly, for preventing side-effects of ifosfamide on the urinary system (haemorrhagic cystitis, etc.).     

Structural and conformational aspects of the anticoagulant and anti-thrombotic activity of heparin and dermatan sulfate

Heparin and other iduronic acid-containing glycosaminoglycans (GAG) such as dermatan sulfate exert their anticoagulant properties primarily by accelerating the rate of inhibition of the natural protease inhibitors antithrombin III (AT, which inhibits both factor Xa and thrombin) and heparin cofactor II (HCII, which selectively inhibits thrombin). Although AT and HCII are structural homologs, only heparin binds to AT, and HCII has different binding sites for heparin and dermatan sulfate. Whereas the binding site of heparin for AT is a unique pentasaccharide sequence contained in only about one third of the chains of this GAG, HCII-binding sequences of heparin and dermatan sulfate are less specific and contained in practically all the GAG chains. Protein binding and associated biological activities of heparin and dermatan sulfate are modulated by the "plasticity" of their iduronic acid residues due to the availability of up to three equienergetic conformation among which the protein selects the one favouring the most stable complex. Glycol-splitting of nonsulfated uronic acid residues, a device for generating flexible joints along the GAG chains, has different effects on different binding domains. Whereas it inactivates the binding site for AT causing a drop of the anticoagulant activity, it enhances the HCII-associated activity of both heparin and dermatan sulfate.     

免疫印迹法分析AFP糖链及其临床意义

本文采用免疫印迹法分析扁豆凝集素与AFP糖链的结合能力,鉴别诊断肝细胞癌。对32例肝细胞癌、14例慢性肝炎、肝硬化患者的AFP逆行了分析,以扁豆凝集素结合型AFP部分大于总量的25%作为诊断肝细胞癌的标准,其中阳性率为100%(32/32),正确率为73.9%(34/46)。     

T-lymphocytic infiltrate in canine mammary tumours: clinic and prognostic implications

In recent years, considerable progress has been made in understanding the role of the immune system in tumour progression. However, in canine mammary tumours (CMT), the prognostic value of T-lymphocytes is not established. The aims of the present study were to characterize T-lymphocytic infiltrate in 57 canine mammary tumours (21 benign and 36 malign), by immunohistochemical detection of CD3 antigen, and to determine its association with several clinicopathological parameters and overall survival. CD3+ positive cells were counted in 10 high-power fields within the tumour (i.e. The tumour-infiltrating T-lymphocytes, TIL), in the peripheral area of the tumour and in the adnexal non-tumoural mammary gland. CD3(+) TILs were significantly more frequent in benign than in malignant tumours (p<0.001). Conversely, peripheral CD3(+) TILs were significantly more frequent in malignant than in benign neoplasias (p<0.001). For CD3(+) T-lymphocytes in the adnexal non-tumoural mammary gland, there was no statistical difference in their frequency between benign and malignant tumours. On survival analysis, there was a tendency towards an association of a higher number of CD3(+) TILs and a shorter overall survival (p=0.08). Interestingly for CD3(+) T-lymphocytes in the adnexal non-tumoural mammary gland, a statistically significant relationship was observed, with a higher number of lymphocytes conferring a reduced overall survival (p=0.045). Further studies will be required to better understand the biological implications of the current findings.     

Decorin from different bovine tissues: study of glycosaminoglycan chain by PAGEFS

The sulphation pattern of glycosaminoglycan (GAG) plays a critical role in biological functions of proteoglycans. In this study, we showed that decorins from different bovine tissues present specific sulphation pattern coupled with peculiar biological activity. In order to elucidate chemical structure of decorin glycosaminoglycan chains, we improved an electrophoretic method to analyse fluorescent disaccharides from dermatan/chondroitin sulphate GAG chains. The disaccharide separation is based on minigels, and this technique was able to define the polysaccharide chain composition in terms of sulphated and not sulphated disaccharides. This approach allowed not only the measurement of few picomoles of material, but it also permits a rapid qualitative analysis of the GAG chains. Data obtained by PAGEFS indicate that the sulphation pattern of GAG is tissue specific and this finding may explain the different binding properties to von Willebrand factor of decorins.     

Local and Systemic Effects of Commonly Used Cutaneous Agents: Lifetime Studies of 16 Compounds in Mice and Rabbits

Abstract A number of commonly used external agents were applied repeatedly on the skin of the mouse and rabbit. These chemicals, components of commercially available products included 4-hydroxyanisole, benzalkonium chloride, bromodeoxyuridine, iododeoxyuridine, 5-nitroacenaphthene, N-N-diethyltoluamide, hexachlorophene, para-aminobenzoic acid, benzophenone, isopropylmyristate, pyrogallol, resorcinol and ethylhexanediol. Also tested were a commercial hairspray, a dandruff shampoo and an anti-dandruff agent. Local and systemic changes were studied and the tumour incidence compared with that of an effective carcinogen, 9,10-dimethylbenz(a)anthracene. The mice showed no local toxic changes or tumour formation and the systemic tumour incidence, i. e., tumours of the liver, lungs, lymphatic system and other organs, was similar to that of control animals. In rabbits, a number of proliferative, benign and malignant ear tumours were observed in the positive controls, thereby demonstrating the efficiency of this model. Local toxic changes were seen in benzalkonium- and hexachlorophene-treated animals, but no skin tumours. Four animals had uterine tumours. In addition, one lung and one kidney tumour, unrelated to the method of treatment, were seen.     

Characterization of chondroitin/dermatan sulfate proteoglycans synthesized by bovine retinal pericytes in culture

Pericytes associate with the outside of endothelial cells in microvessels. Previous studies have shown that these cells synthesize glycosaminoglycans (GAGs) but the nature of the core proteins to which these GAGs are attached is unknown. In the present study, cultured bovine retinal pericytes were metabolically labeled with [(3)H]glucosamine, [(35)S]sodium sulfate or (35)S-labeled amino acids and the proteoglycans synthesized by these cells were purified by DEAE-Sephacel ion exchange and molecular sieve Sepharose CL-4B chromatography. Separated proteoglycans were digested with papain, heparitinase or chondroitin ABC lyase and the GAGs characterized by Sepharose CL-6B chromatography. Proteoglycans were also assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis before and after digestion with chondroitin ABC lyase. Pericytes predominantly synthesize and secrete chondroitin or dermatan sulfate proteoglycans (CS/DS PGs) rather than heparan sulfate proteoglycans (HSPGs). Two subclasses of CS/DS PGs are synthesized by pericytes; one is a high M(r) subclass with high charge density. This subclass eluted at the void volume of a Sepharose CL-4B molecular sieve column, was susceptible to chondroitin ABC lyase, and contained core proteins of ca. 550 and 450 kD which were recognized by antibody to versican. The other major subclass eluted at a K(av) ca. 0.45 on a Sepharose CL-4B molecular sieve column, was susceptible to chondroitin ABC lyase, and contained core proteins recognized by antibodies to either biglycan or decorin that separated as a broad band of ca. 50 kDa in SDS-PAGE. A small amount of HSPG was also synthesized by these cells and could be separated from the CS/DS PGs by DEAE-Sephacel chromatography using a linear gradient of 0.1-0.7 M NaCl. Release of GAG chains by protease digestion indicated that the length of GAG chains was approximately M(r) 45000 in biglycan and decorin, approximately M(r) 48000 in the small amount of HSPGs and approximately M(r) 66000 in versican. These proteoglycans resemble those synthesized by vascular smooth muscle cells but differ markedly from those synthesized by vascular endothelial cells.     

A rare case of renal and uterine clear cell carcinoma: Which is the primary tumour?

Renal cell carcinoma to the uterus is rare. We describe a 52-year old lady who presented with progressive abdominal distension and computerized tomogram scan of the abdomen showing two pathologies; uterine and right renal tumour. It was initially thought to be two distinct tumours (double pathology). Radical nephrectomy and total abdominal hysterectomy with bilateral salpingo-oophorectomy was performed concurrently. Histopathological evaluation of the uterine and right renal tumour however surprised us with a rare form of metastasis from a renal tumour to the uterus. The method of differentiating primary renal cell carcinoma with uterine metastasis, from primary uterine carcinoma with renal metastasis, via immunohistochemistry, is described.     

Immunolocalisation of Cytochrome P‐450 3A Enzymes in Human Breast Carcinoma: Relationship with Tumour Differentiation and Steroid Receptors

Abstract: Cytochrome P‐450 3A enzymes belong to the most abundant subfamily of the cytochrome P‐450 system. They are predominantly found in the liver where they metabolize numerous drugs and endogenous substances such as oestrogens. However, they are also expressed by normal and tumoural extrahepatic tissues. Accordingly, immunolocalization was assessed in malignant breast tumours (n=32) and normal counterparts, by using a monoclonal antibody that recognizes all human CYP3A proteins. We investigated a potential relation between expression of CYP3A protein expression, the degree of tumour differentiation assessed by the histological grade and the proliferation index assessed by Ki‐67 immunostaining. Immunodetection of CYP3A was observed in 27 of the 32 tumours analyzed (84%). A focal staining was also observed in the adjacent normal breast tissue in 33% of the samples, but expression was always fainter than in tumours. A significant negative association was found between CYP3A and the proliferation index, but there was no relation with receptor status or tumour differentiation. While CYP3A protein expression can be found in normal breast tissues, these data highlight higher and more frequent CYP3A in malignant breast cells. Such expression in malignant breast cells appears inversely related to the proliferation index whereas no relation is found with tumour differentiation.