膀胱癌的凝集素表达与组织学分级及预后的关系

选取七种生物素化凝集素采用ＡＢＣ法对１３例正常膀胱粘膜和１０２例膀胱移行细胞癌进行了免疫组化研究，并分析其组织学分级及预后的关系。结果：肿瘤的病理级别与ＣｏｎＡ或ＷＧＡ的表达率呈正相关，与ＳＪＡ的表达率呈负相关；膀胱癌预后不佳组ＣｏｎＡ及ＷＧＡ的强阳性率在８０％以上，而ＳＪＡ受体对９５％病例缺失；５年死亡组的ＣｏｎＡ和ＷＧＡ几乎全部强阳性表达，而ＳＪＡ则９０％以上呈阴性反应。提示ＣｏｎＡ、ＷＧＡ和ＳＪＡ是与膀胱癌分级及预后密切相关的有价值的标记物。     

Modifications of surface carbohydrates on bovine spermatozoa mediated by oviductal fluid: a flow cytometric study using lectins

The objective of the present study was to characterize and quantify changes in exposed saccharide residues of bovine sperm during capacitation in oviductal fluid (ODF) using flow cytometry (FC). Bovine sperm were incubated with 0% or 50% non-luteal ODF for 30 min or 3.5 h. After incubation, sperm were labelled with 11 fluorescein isothiocyanate-labelled lectins and evaluated for lectin binding with FC. Furthermore, inhibiting sugars were used to determine specificity of lectin binding to oligosaccharides on the sperm surface. After 30 min incubation, there was a 91% decrease in fluorescence intensity of labelled sperm incubated in WGA, a 76% decline for Con A, 75% decline for BS-I and a 36% decline for DBA. These differences remained approximately the same over the 3.5-h incubation. Interestingly, although there was no reduction in UEA-I binding at 30 min, a significant reduction (23%) was observed at 3.5 h. Con A fluorescence was mostly inhibited with either alpha-d-glucose or alpha-d-mannose (86% and 90% respectively). BS-I fluorescence was reduced after prior incubation of the control samples with N-acetyl-galactosamine and galactose by 74% and 80% respectively. After prior incubation with N-acetyl-galactosamine DBA fluorescence reduced by 18% in the control samples. With UEA-I no fluorescence reduction was observed after prior incubation with l-fucose. We have demonstrated that capacitation of bovine sperm in ODF is accompanied by a quantitative reduction in individual lectin binding sites. These modifications may be crucial to the subsequent signalling events involved with sperm-zona binding, zona penetration or interaction with the oolema.     

Lectins in diagnosis of bladder carcinoma.

With the purpose of studying changes in the expression of glycoconjugate structures in nonmalignant and cancerous lesions of urothelium the lectins ConA, TKA, PNA, DBA, STA, LFA, UEA, MPA, RCA, LCA, GSA1, SBA, GSA2, WGA, PHA and Lot were tested in formalin-fixed, paraffin-embedded tissue sections of (1) cold biopsies from normal urothelium and bladder cancer of different grades (G1-G3) in humans, (2) normal transitional epithelium and N-butyl-N(4-hydroxybutyl)nitrosamine (BBN)-induced bladder cancer in animal experiments (Wistar rat), and (3) human transitional cancer cell line HT 1376. In human urothelium TKA and SBA were positive markers demonstrating positive staining reactions in all tumor grades without binding to normal epithelium. They stained also the human transitional carcinoma cell line HT 1376 (G3). In Wistar rats DBA, ConA, LCA, SBA, GSA2 and WGA had a specific affinity to BBN-induced carcinoma. Findings of positive lectin marker in transitional cell cancer may offer progress in diagnostics and therapy.     

早早孕人子宫蜕膜及绒毛膜凝集素的组化分析

以凝集素作为组化探针研究早早孕期人子宫蜕膜、绒毛膜表面糖复合物的变化。结果表明：６种凝集素在早早孕期与子宫蜕膜及绒毛膜结合显示不同变化。其中刀豆凝集素（ＣｏｎＡ）、花牛凝集素（ＰＮＡ）、双花藕豆凝集素（ＤＢＡ）与子宫蜕膜结合随妊娠无数增加着色逐渐减弱。荆豆凝集素（ＵＥＡ－Ⅰ）与子宫蜕膜未见着色。麦胚凝集素（ＷＧＡ）、蓖麻凝集素（ＲＣＡ－Ⅰ）与子宫内膜在早早孕期着色程度未见有变化。ＰＮＡ、ＣｏｎＡ在与绒毛滋养层细胞结合随妊娠天数增加着色逐渐增强。ＵＥＡ－Ⅰ和ＤＢＡ与滋养层细胞在各期中未见结合。     

The study of the ABH isoantigen of bladder tumors

ABH isoantigens in 82 cases with bladder tumor of various grades and stages were investigated by the avidin-biotin-peroxidase complex (ABC) method. Correlation of ABC expression for isoantigens with pathological findings on the grading and staging system of the bladder tumors were as follow: Of 60 cases of low grade tumors, 34 (56.7%) were positive and of 22 cases of high grade tumors, 21 (95.5%) were negative. Of 63 cases of low stage tumors, 35 (55.6%) were positive and all 19 cases of high stage tumors, (100%) were negative. The isoantigens in the tissue of the bladder tumors tended to disappear as the histologic anaplasia of the tumor progressed. There was a high correlation between the histological grade, stage and ABC expression (p less than 0.01). In a follow-up study of 38 patients with low grade, low stage, initial transitional cell carcinoma, 9 of the 24 patients positive for ABC expression (37.5%) showed recurrence during the 1 to 6 year follow-up period, whereas 10 of the 14 patients negative for ABC expression 10 (71.4%) did. There was a high correlation between the recurrence rate and ABC expression (p less than 0.05). Therefore, the analysis in the bladder tumors may be valuable for prediction of malignant potential, especially for the low grade, low stage tumors.     

Lectin binding in immunohistologically AFP/HCG positive and negative testicular carcinoma

The morphologic display of testicular cancer is a heterogenous cellular pattern. A biological heterogeneity is also true for the expression of tumor markers. The biosynthesis of tumor marker proteins alpha-fetoprotein (AFP), ferritin, Schwangerschaftsprotein (SP 1) glycoprotein, tissue polypeptide antigen and of hormones (beta-human chorionic gonadotropin (HCG) = significantly present in nonseminoma germ cell tumors--does, however, define only a small number of cancer cells. To better visualize the majority of cancer cells, lectin binding was studied. During the oncogenic transformation a distinct change of cell membrane glycoproteins has been observed. Reactions of WGA/PNA lectins which get attached to glycoproteins with cancer tissue sample from seminomas (n = 20) and nonseminomas (n = 20) were analyzed. The results were correlated to AFP/beta-HCG positive (negative) immunohistology to establish further subgroups of biological homogeneity. The binding of WGA lectin appears relatively more frequent in both seminoma and nonseminoma than that of PNA. Lectin binding of WGA and/or PNA can be stained in 3/11 AFP- and beta-HCG-negative nonseminoma tissues while lectin staining is positive in 7/18 beta-HCG-negative seminomas. The fact that lectin binding is dependent on the spermatozoogenesis and on androgens in normal testis tissues asks for more detailed studies in this field.     

Expression of ras p21 oncogene product on human bladder tumors

The immunohistochemical localization of ras p21 oncogene product was examined in human bladder cancer. These bladder cancers consist of 52 superficial bladder tumors and 10 cases of invasive tumor. Five (9.6%) of 52 superficial tumors were demonstrated to be positive for ras p21 product. Two (20%) of 10 cases which showed deep tumor infiltration were demonstrated to be positive for ras p21 oncogene product. When we analyzed the incidence of positive staining for ras p21 with regard to histological grade of bladder cancer, 2 of 30 (7%) patients with G1, 4 of 26 (15%) with G2, and 1 of 6 (17%) with G3 had positive staining for the ras p21 oncogene product. One of five patients who showed positive staining on tumors had tumor recurrence 3 months later. The remaining 4 patients positive for the ras p21 oncogene product had no tendency for recurrence in 11-19 months' follow-up. Therefore, the incidence of detection of ras p21 oncogene product on superficial and deep infiltrating bladder tumor was similar and there was no significant correlation between the incidence of positive ras p21 staining and depth of invasion of bladder tumors or histological grade.     

Separation of osteoclasts by lectin affinity chromatography

Newborn rat calvaria bone cells obtained by digestion were fractionated on columns of wheat-germ agglutinin (WGA) sepharose 6MB for osteoclast isolation. The initial nonspecific binding cells which were passed through the WGA sepharose column by a buffer acquired a high enzyme activity of alkaline phosphatase, but not that of acid phosphatase. However, elution of cells using a buffer with the addition of N-acetyl-D-glucosamine resulted in a high acid phosphatase activity but no alkaline phosphatase activity. The former WGA binding negative fraction enriched osteoblasts averaging 30 microns in size. The latter WGA binding positive fraction enriched osteoclasts ranging from 20 microns to 60 microns in size. The electron-microscope clearly demonstrated the cellular details of osteoclasts. Isolated cell counts showed a ratio of six to four. These results indicate that our method of osteoclast isolation is simple and useful in lectin affinity chromatography because all cells have sugar moieties on their surface and the binding of osteoclasts can be reversed by the addition of specific lectin-binding sugars to the eluting buffer.     

Surveillance of stage O, grade I bladder cancer by cytology alone--is it acceptable?

Although the high incidence of subsequent tumors is well established there is accumulating evidence that few cases of low grade, low stage transitional cell carcinomas will progress in stage. Since the purpose of intensive endoscopic monitoring following initial tumor resection is to detect potentially lethal new tumors as soon as possible, we reviewed retrospectively the course of 36 patients with an initial grade I, stage O (Ta) transitional cell carcinoma to determine whether cytology was capable of achieving this goal. Of these patients 10 (28 per cent) had a subsequent tumor of a higher grade or stage. Cytology performed at or before recurrence was positive in 8 patients (80 per cent) and 2 (20 per cent) had grade II, noninvasive transitional cell carcinoma with negative cytology. Subsequent tumors in these 2 patients have been grade I. All 36 patients are alive. Given the patient inconvenience, expense and risk of infection of cystoscopy compared to cytology, this retrospective review suggests that when an experienced cytopathologist is available patients with grade I, noninvasive transitional cell carcinoma may be monitored primarily by urinary cytology with less frequent endoscopy. A prospective study must be performed to confirm this approach.     

Control group and maintenance treatment with bacillus Calmette-Guerin for carcinoma in situ and/or high grade bladder tumors

PURPOSE: Intravesical instillations of bacillus Calmette-Guerin have demonstrated satisfactory results in the treatment of vesical carcinoma in situ and high grade superficial bladder tumors. We designed a protocol to evaluate the decrease in tumor recurrence with maintenance therapy. MATERIALS AND METHODS: Between June 1989 and May 1995 an initial course of 6 intravesical instillations of Connaught strain bacillus Calmette-Guerin was administered in patients with carcinoma in situ and/or high grade superficial bladder tumors. Six months later 131 disease-free patients were randomly assigned to a control group or a maintenance therapy group that received 6 instillations every 6 months (6 x 6) for a 2-year period. RESULTS: Of the 126 evaluable patients at a mean followup of 79 months there were no significant differences in recurrence nor progression. A total of 16 patients (26.2%) in the control and 10 (15.1%) in the maintenance group had superficial relapse at a mean of 24 and 20 months, respectively (p = 0.07). Eight patients underwent radical cystectomy due to bladder contraction in 1, high grade superficial recurrence in 4 and disease progression in 3. Of the 65 patients on maintenance therapy 22 (33.85%) completed the planned 2-year treatment. CONCLUSIONS: Six-month maintenance therapy in patients treated initially for carcinoma in situ and/or high grade superficial bladder tumors who are disease-free at 6 months did not significantly decrease recurrence or progression.     

Lectin-Binding Sites in Testis of Men with Acrosomeless Round-Headed Spermatozoa/Testikuläre Lektinbindungsstellen bei Männern mit akrosomenlosen Rundkopfspermatozoen

We report herein about lectin histochemistry of seminiferous epithelia in two infertile men with exlusely acrosomeless round-headed spermatozoa. FITC-conjugated lectins (ConA, PNA, RCA II, WGA) have been employed on tissue sections of Bouin fixed testicular biopsies. RCA II gave a dot-like fluorescence of the acrosomal region and WGA gave a cap-like acrosomal fluorescence of spermatids. PNA-a marker of acrosomal differentiation-failed to stain spermatids. The binding of ConA to germ cells was not influenced by this syndrome. In conclusion, the syndrome of acrosomeless round-headed spermatozoa is associated with selective perturbations of testicular lectin-binding sites. They might contribute of the inability of sperm cells to adhere to and penetrate into ova.     

Capacitated and acrosome reacted spermatozoa of goat (Capra indicus): a fluorescence and electron microscopic study

Summary. Plasma membrane alterations accompanying in vitro capacitation and acrosome reaction of goat spermatozoa were studied using lectin labelling, scanning electron microscopy, and freeze-fracture methods. Fluorescein isothi-ocyanate linked lectins namely; Canavalia ensiformis (ConA), Maclura pomifera (MPA), Arachis hypogaea (PNA), Glycine max (SBA) and Triticum vulgaris (WGA) agglutinin were used to examine the distribution of surface carbohydrates during these two events. The head and the sperm tail reveal altered lectin labelling features after capacitation and acrosome reaction. After capacitation the surface coat components for MPA, SBA, and WGA are shed from the spermatozoon head. ConA receptors on the head are retained after capacitation but are partially shed in the acrosome reacted spermatozoa. SBA receptor sites appear on the sperm tail of the capacitated spermatozoa. Unusual morphological changes attending capacitation involve the sperm tail-end which develops a novel entity, which we have termed 'spatula'. The 'spatula' shows strong binding with ConA and WGA only. In the acrosome reacted spermatozoa the spatulated tail-end unwinds with a concomitant loss of lectin labelling. Highly ordered membrane particles, 'ladders' of the middle piece of the epididymal sperm tail, disappear and IMP clearings appear on the middle piece and in the spatulated ends of the capacitated spermatozoa. But in the acrosome reacted sperm IMPs reappear and are randomly disposed on the middle-piece and are clustered in small patches on the principal-piece. IMP free areas appear on the plasma membrane covering the acrosome and the outer acrosomal membrane (OAM) of the capacitated spermatozoa. The plasma membrane and OAM fuse at multiple foci and appear as acrosomal 'ghosts' which remain associated with the sperm head even after acrosome reaction.     

Grade progression and regression in recurrent urothelial cancer

PURPOSE: Recurrent urothelial cancers are reported to have characteristics similar to those of the primary tumor, with 10% to 25% of low grade tumors recurring as high grade disease. We determined how often grade progression and regression occur and whether abnormalities in p53 protein expression in original tumors are preserved in recurrences. MATERIALS AND METHODS: Two groups of patients treated for recurrent stages Ta/T1 urothelial bladder cancers with at least 1 tumor-free examination between the index and recurrent tumors were reviewed. Group 1 included 115 patients in whom the first available tumor was compared with the last recurrence and group 2 included 42 in whom the initial tumor was compared with the first recurrence. Immunohistochemical analysis of p53 expression was performed on a subset of 34 tumor pairs. RESULTS: In group 1, 33 grade 3 tumors (45%) recurred as grade 1 or 2 tumors, while 9 of 82 grades 1 and 2 tumors (11%) recurred as grade 3 tumors. Five of 7 group 2 grade 3 tumors (71%) recurred as grade 1 or 2 disease, while 1 of 35 grades 1 and 2 tumors (3%) recurred as grade 3 disease. In the 34 pairs studied immunohistochemically 6 of 14 grade 3 tumors recurred at lower grades. Nuclear p53 over expression occurred in 21 index tumors (12 of 14 grade 3, 8 of 17 grade 2 and 1 of 3 grade 1) and in 9 recurrences (6 of 10 grade 3, 2 of 17 grade 2 and 1 of 7 grade 1). Only 7 of 21 p53 positive and 2 of 12 p53 negative index tumors were p53 positive on recurrence. CONCLUSIONS: While progression from low to high grade occurred in less than 15% of patients, grade regression was observed in almost 50%. The loss of p53 positivity in regressing tumors indicates that these recurrences are molecularly distinct from the corresponding initial tumor.     

Superficial bladder tumor. Aspects of clinical progression

The clinicopathologic staging of bladder cancer is the main prognostic aid in superficial tumor since it has close relation to the degree of differentiation of the tumor and the observed degree of its progression in the 238 cases of this series. The survival rate of the patients was related to the depth of penetration and degree of histologic differentiation of the tumor. Thus both the stage and the grade of the tumor together had prognostic value. Primary clinical staging correlated well with the size of the initial lesion, but not with the location, number of primary lesions or subsequent recurrences, or time of the appearance of progression of the primary lesion. Almost one third of the patients with superficial bladder cancer died from bladder cancer. Two thirds of the cases with known pathologic stage at autopsy showed extravesical extension of the lesion. Prior external radiation to the superficial tumors apparently did not affect progression of the primary lesion.     

Intravesical bacillus Calmette-Guerin instillation therapy in superficial bladder cancers--clinical study on prognostic factors

Intravesical instillation of Tokyo 172 strain bacillus Calmette-Guerin (BCG) was performed on 137 patients with superficial bladder cancer (Ta and T1) after transurethral tumor resection as a prophylaxis against tumor recurrence. The recurrence rate of tumors was compared with that of historical controls and was estimated by the person-years method. There were statistically significant decreases in recurrent tumors following BCG therapy. To clarify the efficacy of intravesical BCG therapy, prognostic significance of several factors were evaluated in 90 patients with initial bladder cancer treated with TUR and BCG instillation, and compared to those of controls. Prophylactic effects were statistically better for those with multiple tumors, grade 3 lesions or Ta lesions than for control patients. No correlation between purified protein derivative responsiveness and favorable results could detected. There were no marked side effects or fatal complications of BCG therapy during the observation periods. Our results suggest that BCG is able to change the biological behavior of superficial bladder cancers with multiple, high grade or low stage lesions. The intravesical BCG instillation seems to be effective and safe as a prophylaxis against the recurrence of superficial bladder tumors.     

Lichtmikroskopischer Nachweis von Lektin-Rezeptoren an der Oberfläche humaner Spermatozoen/Demonstration of Lectin-receptors on the Surface of Human Spermatozoa by Light-microscopy

The topography of lectin binding sites on human spermatozoa has been examined by using particular series of peroxydase labeled lectin-konjugates (GS-1, GS-2, WGA, PNA, SBA, DBA, BPA, MPA, UEA-1, LPA, Con-A) which demonstrated typical patterns of lectin-receptor distributions concerning acrosomal and equatorial as well as postacrosomal areas.     

Blood group isoantigen and lectin binding studies in the human bladder cancer

The purpose of this study is to investigate changes of glycoconjugates in the cancer cells of the urinary bladder by means of immunohistochemical methods. The normal bladder epithelium, cancerous lesions, and non-malignant epithelia of the tumor bearing bladder were examined by staining by avidin-biotin-peroxidase complex methods using blood group isoantigens (BGA) and lectins. The materials were obtained from 48 cystectomy specimens in our hospital in these seven years. Anti-A, B and H monoclonal antibodies were used for detecting BGA. GSI-A4, UEA-1, LTA-M, BPA, DBA and PNA were used as probes for lectins. The changes of glycoconjugates in the cancer cells of the bladder and in non-malignant epithelia of the tumor bearing bladder were studied by using six kinds of lectins. Each lectin binding rates of the primary lesions and the metastasized lesions was comparatively investigated. Positive rates for BGA of the high grade tumor was lower than that of the low grade. In the high grade tumor, GSI-A4, PNA and BPA showed high binding rates, while the DBA binding rate was low. The correlation between the histopathological grading of bladder tumor and the changes of glycoconjugates in the cells was suggestive of cancerous process.     

Testicular Lectinhistochemistry in the Sertoli-Cell-Only-Syndrome/Testikuläre Lectinhistochemie beim Sertolizell-Syndrom

Glycoconjugate expression was studied in tissue specimens of the Sertoli-cell-only-syndrome by lectinhistochemistry. Both ConA and WGA marked the cell surface of Sertoli cells fainty. UEA I labelled their nucleoli. PNA and RCA failed to stain Sertoli cells. These staining patterns resembled those of normal human testis. The thickened tubular walls in the Sertoli-cell-only-syndrome were abundant in binding sites for WGA, PNA, UEA I, and RCA. ConA stained the outer part of the wall heavily, but failed to react with the adluminal part. It is suggested, that the Sertoli-cell-only-syndrome is accompanied by a selective loss of mannosyl residues in the adluminal part of thickened tubular walls.     

Carbohydrate residues in non-malignant prostatic epithelium as revealed by lectins

Summary Non-neoplastic prostatic epithelium from 39 patients obtained at transurethral resection for outflow tract obstruction and 5 normal prostates from men under 35 years of age obtained at postmortem were formalinfixed and paraffin-embedded. The distribution of 8 lectin receptors were studied using a peroxidase anti-peroxidase method and an avidin-biotin method. Con A, WGA, and PNA bound to most epithelial cells. Con A,and WGA also showed major stromal binding. Approximately 5% to 10% of cells bound UEA1, GS1, DBA, and BPA. No major differences in lectin receptor expression were observed between normal and hyperplastic epithelium with either of the immunohistochemical techniques except that hyperplastic cells stained more strongly than normal epithelium.     

Histological evaluation of superficial bladder tumors treated by Nd-YAG laser and transurethral resection.

Based on 3 study groups we have tried to analyze some concepts concerning the treatment of superficial bladder tumors by the Nd-YAG laser. In the first group of 34 superficial bladder tumors, we evaluated the accuracy of cold-cup biopsy in staging the tumor before treatment with the laser (100% for grade and 90% for stage). The possibility of identifying the tumoral grade in the lasered mass is 73%. In the second group of 53 random superficial bladder tumors we compared the effects of laser and transurethral resection (TUR) with regard to recurrence and progression after a 3-year follow-up: no significant differences are found between the treatments. In the third group the histologic changes in the bladder wall after treatment with the laser or TUR were evaluated by cold-cup biopsy (31 lasered cases and 24 resected cases) and by resector loop biopsy (11 lasered cases and 11 resected cases) between 12 and 24 months after treatment, without significant differences in terms of fibrosis.