Stereological analysis of nasal mucosa. III. Stepwise alterations in cellular and subcellular components of pseudostratified, metaplastic and dysplastic epithelium in nickel workers

Nasal mucosal biopsies taken primarily for diagnostic purposes were graded as pseudostratified, metaplastic (consisting of stratified cuboidal, non-keratinizing and keratinizing stratified squamous epithelium) or dysplasic, and were analysed by simple stereological procedures at the electron microscopic level. For biological and methodological reasons the evaluation was focused on the basal cells of the epithelium. The transition of pseudostratified to metaplasic and dysplasic epithelium was accompanied by significant alterations in the karyoplasm and cytoplasm. The main changes in the karyoplasm were a gradual reduction of heterochromatin and an enlargement of the nucleoli. In the cytoplasm the main changes involved: (i) the mitochondria, with a threefold increase in size and a corresponding reduction in number. In addition the surface density of their cristae was reduced by 32%. (ii) The surface density of rough endoplasmic reticulum, which increased by 100%, (iii) the volume fraction of tonofilaments which decreased by 60%, and (iv) the number of desmosomes which increased by 60%. The morphologic changes, especially those involving the mitochondria and endoplasmic membranes, suggest that biochemical differences also exist between the original, and metaplasic and dysplasic epithelia. The stepwise alterations of several independent cellular and subcellular components support the hypothesis of a gradual transformation of pseudostratified epithelium to stratified squamous epithelium and dysplasia.     

Usefulness of AgNOR technique and CEA expression in atypical metaplastic cells from cervical smears

OBJECTIVE: To evaluate the proliferating capacity of atypical metaplastic cells in cervical smears by AgNOR technique and image analysis and investigate the probable relation with squamous intraepithelial lesions (SIL) using immunocytochemical assay for carcinoembryonic antigen (CEA). STUDY DESIGN: Eight cervical smears were stained with Papanicolaou stain for diagnosis of atypical metaplastic cells. After removal of the stain the smears were processed with a silver colloidal solution and the AgNOR area determined by image cytometry. Differences in the mean AgNOR protein area between reactive metaplastic cells and controls were tested by Student's t test. The CEA was investigated by immunocytochemical staining in smears with atypical metaplastic cells and smears from high-grade squamous intraepithelial lesions (HSIL). RESULTS: The mean AgNOR areas from atypical metaplastic cells (4.55, 6.66, 4.68, 5.30, 4.97, 6.20, 6.28, and 7.35) were significantly greater those that of intermediate metaplastic cells and cells from low-grade squamous intraepithelial lesions (LSIL) (0.77, 0.99, and 0.82, respectively). The atypical metaplastic cells showed values of mean AgNOR area intermediate between that of basal cells (3.28) and HSIL cells (7.73) or neoplastic cells (16.12). The CEA was strongly expressed by the atypical metaplastic cells. CONCLUSION: The expression of CEA in the atypical metaplastic cells underlies the probable relation to SIL. Although the organizer region areas raised high values, it would be necessary to use a greater number of cases to define whether the AgNOR area is indicative of the proliferative activity of the atypical metaplastic cells.     

Evidence for a role of the monoclonal antibody E48 defined antigen in cell-cell adhesion in squamous epithelia and head and neck squamous cell carcinoma

Monoclonal antibody (MAb) E48 recognizes a 20- to 22-kDa antigen expressed by human squamous and transitional epithelia and their neoplastic counterparts. Histochemical examination of these tissues revealed distinct surface labeling with MAb E48. To investigate the subcellular localization of the E48 antigen we have performed electron microscopical analysis. In cells of normal oral mucosa, the E48 antigen was expressed on the plasmalemma, particularly associated with desmosomes, suggesting involvement of the E48 antigen in intercellular adhesion. Furthermore, the level of expression of the E48 antigen appeared to be influenced by the cellular organization. In squamous cell carcinoma (SCC) cell lines grown in vitro as subconfluent monolayer cultures, the E48 antigen expression was low. However, E48 antigen expression increased when SCC cells were grown to confluency. E48 antigen expression was similarly high when SCC cell lines were cultured under conditions promoting three-dimensional growth either as colonies within floating collagen gels or as xenograft in tumor-bearing nude mice. Further evidence for the involvement of the E48 antigen in cell-cell adhesion was found when SCC cells were grown within collagen gels in the presence of MAb E48: no spherical colonies were formed, but cells grew out to colonies composed of single cells. Moreover, in this culture system the percentage of SCC cells growing out to colonies was decreased by the presence of MAb E48. These findings indicate that the E48 antigen is involved in the structural organization of squamous tissue and might have a role in intercellular adhesion.     

Ⅲ期非小细胞肺癌患者精确放疗前后血清CEA、SCC、NSE水平变化及与放疗疗效的关系研究

目的:研究Ⅲ期非小细胞肺癌(NSCLC)患者精确放疗前后血清癌胚抗原(CEA)、鳞状细胞癌相关抗原(SCC)、神经元特异性烯醇化酶(NSE)水平变化及与放疗疗效的关系。方法:选择2014年1月到2016年12月在亳州市人民医院肿瘤科就诊的60例Ⅲ期NSCLC患者纳入此次研究,其中鳞癌14例,腺癌26例,腺鳞癌20例。所有患者均实施4周的精确放疗,放疗后肿瘤标记物水平降低43例,升高17例。根据放疗疗效将患者分为有效组39例,无效组21例。对比不同病理类型的Ⅲ期NSCLC患者CEA、SCC、NSE水平,不同疗效组放疗前后CEA、SCC、NSE水平,并分析患者的肿瘤标记物水平变化与放疗疗效的关系。结果:腺癌Ⅲ期NSCLC患者的CEA、NSE水平高于鳞癌及腺鳞癌者,且腺鳞癌者又高于鳞癌者;SCC水平低于鳞癌及腺鳞癌者,且腺鳞癌者又低于鳞癌者(P0.05)。放疗后有效组CEA、SCC、NSE水平均低于放疗前和无效组,而无效组CEA、SCC、NSE水平高于放疗前(P0.05)。肿瘤标记物水平降低者的有效率高于升高者,差异有统计学意义(P0.05)。结论:在实施精确放疗后治疗有效的Ⅲ期NSCLC患者,其血清CEA、SCC、NSE水平均呈现出明显的下降趋势,且与病理类型密切相关,临床上可重点关注上述指标水平,有助于患者的诊疗过程。     

Rare coexistence of keratinizing squamous metaplasia with xanthogranulomatous pyelonephritis. Report of a case with the role of immunocytochemistry in the differential diagnosis

BACKGROUND: Xanthogranulomatous pyelonephritis (XPN), a rare form ofchronicpyelonephritis, is commonly associated with lithiasis and rarely leads to keratinizing squamous metaplasia. Its manifestations closely mimic those of a renal neoplasm, leading to misdiagnosis of malignancy, often resulting in radical nephrectomy. The role of immunocytochemistry in the preoperative cytologic diagnosis is assessed in the present case report. CASE: A 20-year-old male presented with fever and an enlarging mass in the right renal angle. Ultrasonography revealed a heterogeneous mass in the renal pelvis. Fine needle aspiration cytology was advised to rule out malignancy. Aspiration smears from the mass showed many dissociated cells and clusters of them with abundant vacuolated cytoplasm, vesicular nuclei and prominent nucleoli in some cells. Many desquamated metaplastic squamous cells were also seen. The background was predominantly necrotic, with inflammatory cells. The cytologic possibility of XPN with squamous metaplasia vs. renal cell carcinoma was considered. Immunocytochemical markers, epithelial membrane antigen (EMA) and CD68 (histiocytic marker) were used to determine the nature of the suspicious vacuolated cells; these cells were immunoreactive for CD68 and negative for EMA, thus confirming the cytologic diagnosis of XPN with keratinizing squamous metaplasia. CONCLUSION: The case highlights the presence of metaplastic squamous cells in XPN in smears for the first time. Immunocytochemistry is an essential tool in the preoperative cytologic diagnosis of XPN. The patient can be managed conservatively with antibiotics.     

鳞癌细胞增殖能力与岩藻糖基转移酶表达水平及糖蛋白岩藻糖基化修饰有关

岩藻糖基转移酶（fucosyltransferases,FUTs）是一类催化糖蛋白和糖脂发生岩藻糖基化（修饰）酶,主要包括FUT1~FUT9。已有研究证明,很多癌组织中都有不同FUT基因表达升高的现象。本研究证明,表皮鳞癌细胞的增殖能力与几种FUT基因表达水平有关。本文比较研究了人表皮鳞癌A431和SCC12细胞的增殖速度和几种FUT的表达状况,以揭示鳞癌细胞增殖能力与几种FUT基因表达水平的关系。细胞倍增时间结合MTT法揭示,鳞癌A431细胞的倍增时间约为26 h,而鳞癌SCC12细胞的倍增时间约为33 h（P < 0.05）,提示A431细胞增殖速度比SCC12细胞明显加快。与增殖速度一致的是,Western 印迹显示,A431细胞中与DNA合成相关的增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）蛋白表达水平比SCC12细胞高。实时定量PCR（qPCR）检测FUT1-9基因 mRNA转录本,揭示A431细胞中几种FUT基因的mRNA水平均显著高于SCC12细胞。凝集素免疫印迹法和Western 印迹法进一步证明,A431细胞中总蛋白的岩藻糖基化水平比SCC12细胞中的明显升高。敲低FUT4基因表达后,A431细胞中LeY寡糖的表达水平下调,细胞增殖被明显抑制。这些结果证明,较强的表皮鳞癌细胞增殖能力可能与几种FUT基因的高表达,以及糖蛋白的岩藻糖基化（修饰）相关。岩藻糖基转移酶表达水平与临床表皮鳞癌的恶性增生的相关性有待进一步证明。     

Endocervical and metaplastic cells: comparison of endocervical and metaplastic cell number in Papanicolaou smears with and without squamous intraepithelial lesion

OBJECTIVE: To compare, with a retrospective study, endocervical and metaplastic cell number to identify the cytology of squamous intraepithelial lesion (SIL) of the cervix. STUDY DESIGN: Cytology files from March 2000 to March 2003 were reevaluated, and those with SIL were identified. RESULTS: In 8,743 cervicovaginal smears were found 70 (0.8%) SIL smears (39 low grade, 31 high grade) and 140 randomly selected smears without squamous or glandular abnormalities that were not atrophic or unsatisfactory. Endocervical and metaplastic cells counted in these smears and then classified into 5 groups were compared with each other. CONCLUSION: Women with smears containing endocervical cells are not at greater risk for SIL than those with smears lacking these cells. Women with smears containing metaplastic cells are more likely to have SIL than those without metaplastic cells. In addition, high grade lesions are more likely in smears containing more metaplastic cells.     

EGF stimulates Cdc42-dependent translocation of SCC antigen to the plasma membrane

Squamous cell carcinoma (SCC) antigen, including intracellular serine protease inhibitors, is widely used as a laboratory marker for cancers of squamous cell origin. Clinical evidences suggest that increased tissue-expression of SCC antigen predicts an invasive phenotype of cancer cells. Herein, we demonstrated that over-expression of SCC antigen increased the rate of EGF-stimulated cell migration. In the search for the underlying molecular mechanism, we have discovered that SCC antigen was translocated to the plasma membrane upon EGF stimulation and co-localized with polymerized-actin at lamellipodia. We further showed that, co-expression of Cdc42, a downstream target of the EGF receptor, enhanced translocation of the SCC antigen, while co-expression of dominant-inhibitory Cdc42 diminished its translocation. These results suggest that EGF-Cdc42 signal regulates the translocation of SCC antigen to the plasma membrane. Lamellipodia at the leading edge might be a site of action of SCC antigen.     

Squamous cell carcinoma antigen in genitourinary tumors

We have evaluated the serum levels of squamous cell carcinoma (SCC) antigen in patients with adenocarcinoma of the kidney, adenocarcinoma of the prostate, germ cell tumors of the testis, transitional cell carcinoma of the bladder, and SCC of the penis, urethra, and bladder. Serum SCC antigen levels were elevated in 5 of 11 patients (45%) with metastatic SCC of the penis, and in the 3 patients for whom serial determinations were made, the serum levels correlated correctly with the progression of disease or response to treatment. The antigen was elevated in 1 of 3 patients with SCC of the urethra, and 1 apparent false-positive value was observed in a patient with adenocarcinoma of the prostate. Otherwise, no SCC antigen elevations were noted among 10 patients with metastatic adenocarcinoma of the prostate, 8 with metastatic adenocarcinoma of the kidney, 11 with metastatic transitional cell carcinoma of the bladder, 8 with metastatic nonseminomatous germ cell tumors of the testis, and 2 patients with metastatic SCC of the bladder.     

Squamous cell carcinoma antigen is a new member of the serine protease inhibitors

We have cloned cDNA of squamous cell carcinoma antigen. Sequence analysis of the complete 1711 basepairs SCC antigen cDNA revealed that it coded 390 amino acids and no typical signal sequence in the NH2-terminus. Northern blot analysis of human squamous cell poly(A)+ RNA using this cDNA insert as the probe showed a single RNA species of about 1.7 kilobases. The cDNA was expressed in Escherichia coli and the product was detected by immunological methods using antibodies against SCC antigen, indicating that this cDNA encodes the entire SCC antigen sequence. The amino acid homology search revealed that SCC antigen was a member of the serine protease inhibitors family.     

Identification of cultured cell employing specific substance--with special references of "reserve cells" in uterine cervix

Identification of endocervical "reserve cell", which have been regarded as the origin of squamous cell carcinoma of the uterine cervix, was attempted employing immunohistochemically specific substances. The antigenicity of keratin, squamous cell carcinoma antigen(SCC), epithelial membrane antigen(EMA), tissue polypeptide antigen(TPA), vimentin, secretory component(SC) and placental alkaline phosphatase(PLAP) was investigated in histological preparations as well as cultured cells obtained from primary culture of endocervical tissue. The immunohistochemical findings in histological preparations revealed the following: a strongly positive reaction with TPA, a slightly positive reaction with EMA, a very slightly positive with SCC and PLAP, and a negative reaction with keratin, vimentin and SC. Cultured cells were divided into 4 groups according to their morphological characteristics; among these, small rounded or polygonal cells with a centric single nucleus showed similar immunocytochemical reactions to those of "reserve cells" in histological preparations, indicating that "reserve cell" can be growing in culture. The results obtained suggest that immunohistocytochemical specific substances may be useful to identify cultured cells.     

Characteristics of Nasal-Associated Lymphoid Tissue (NALT) and Nasal Absorption Capacity in Chicken

As the main mucosal immune inductive site of nasal cavity, nasal-associated lymphoid tissue (NALT) plays an important role in both antigen recognition and immune activation after intranasal immunization. However, the efficiency of intranasal vaccines is commonly restricted by the insufficient intake of antigen by the nasal mucosa, resulting from the nasal mucosal barrier and the nasal mucociliary clearance. The distribution of NALT and the characteristic of nasal cavity have already been described in humans and many laboratory rodents, while data about poultry are scarce. For this purpose, histological sections of the chicken nasal cavities were used to examine the anatomical structure and histological characteristics of nasal cavity. Besides, the absorptive capacity of chicken nasal mucosa was also studied using the materials with different particle size. Results showed that the NALT of chicken was located on the bottom of nasal septum and both sides of choanal cleft, which mainly consisted of second lymphoid follicle. A large number of lymphocytes were distributed under the mucosal epithelium of inferior nasal meatus. In addition, there were also diffuse lymphoid tissues located under the epithelium of the concha nasalis media and the walls of nasal cavity. The results of absorption experiment showed that the chicken nasal mucosa was capable to absorb trypan blue, OVA, and fluorescent latex particles. Inactivated avian influenza virus (IAIV) could be taken up by chicken nasal mucosa except for the stratified squamous epithelium sites located on the forepart of nasal cavity. The intake of IAIV by NALT was greater than that of the nasal mucosa covering on non-lymphoid tissue, which could be further enhanced after intranasal inoculation combined with sodium cholate or CpG DNA. The study on NALT and nasal absorptive capacity will be benefit for further understanding of immune mechanisms after nasal vaccination and development of nasal vaccines for poultry.     

The Characteristics and Function of S100A7 Induction in Squamous Cell Carcinoma: Heterogeneity,Promotion of Cell Proliferation and Suppression of Differentiation

S100A7 is highly expressed in squamous cell carcinomas (SCC) and is related to the terminal differentiation of keratinocytes. However, its characteristic and function in SCC is not very known. In this present study, we used immunohistochemistry to examine the expression of S100A7 in 452 SCC specimens, including the lung, esophagus, oral cavity, skin, cervix, bladder, and three SCC cell lines. We found that S100A7-positive staining showed significant heterogeneity in six types of SCC specimen and three SCC cell lines. Further examination found that S100A7-positive cells and its expression at mRNA and protein levels could be induced in HCC94, FaDu, and A-431 cells both in vitro and in vivo using immunohistochemistry, real-time PCR, and Western blotting. Notably, the upregulation of squamous differentiation markers, including keratin-4, keratin-13, TG-1, and involucrin, also accompanied S100A7 induction, and a similar staining pattern of S100A7 and keratin-13 was found in HCC94 cells both in vitro and in vivo. Further study revealed that the overexpression of S100A7 significantly increased proliferation and inhibited squamous differentiation in A-431 cells both in vitro and in vivo. Conversely, silencing S100A7 inhibited cell growth and survival and increased the expression of keratin-4, keratin-13, TG-1, and involucrin in HCC94 cells. Therefore, these results demonstrate that S100A7 displays heterogeneous and inducible characteristic in SCC and also provide novel evidence that S100A7 acts as a dual regulator in promoting proliferation and suppressing squamous differentiation of SCC.     

Growth requirements and characterization of rat cervical epithelial cells in culture

The extended culture of rat cervical epithelial cells can be achieved in the absence of a fibroblast feeder layer by utilizing collagen gels and a complex growth medium. The medium contains a 1:1 mixture of RPMI-1640 and Ham's F12 supplemented with 7.5% porcine serum and epidermal growth factor, cholera toxin, transferrin, insulin, and hydrocortisone. Under these culture conditions the cells show rapid log-phase growth and high saturation densities while retaining the ultrastructural characteristics of immature squamous metaplastic cells of the rat uterine cervix even after extended passage. In a manner similar to epithelial cells from a variety of sources, rat cervical epithelial cells form hemicysts at confluence in vitro when cultured on impermeable substrates. The development of these methods for culturing cervical epithelial cells provides an experimental system for the study of factors important in regulating the growth and differentiation of metaplastic squamous epithelial cells.     

Discovery and identification of anti-U1-A snRNP antibody in lung cancer

Autoantibodies are frequently observed in sera of patients with malignancies and generally have been thought to be nonspecific and a reflection of can cer-related general immune system dysfunction. In the previous study, it was reported that autoimmune responses to cell cycle-regulatory proteins and nuclear proteins were present in cancer patients. For example, the antibody against human p53 protein was found in 20%―40% of esophageal carcinoma and oral squa- mous cell carcinoma[1], autoantibo…     

Discovery and identification of anti-U1-A snRNP antibody in lung cancer

There are multiple reports of autoimmune response in patients with lung cancer. To investigate whether a novel autoantibody is present in patients with lung cancer and evaluate its clinical diagnostic and prognostic value, sera from 10 patients with lung cancer and 10 normal individuals were analyzed using immunofluorescence and Western blotting. It was found that one serum sample from the patients with squamous carcinoma gave a fine speckled pattern staining in nucleus and had a high titer antinuclear autoantibody which could recognize 31 kD of nuclear protein isolated from both cancer cells and normal cells. The same patient’s serum was further used to immunoprecipitate the target antigen. The protein bands were excised from the SDS-PAGE gels and were analyzed with a Qstar Pulser I Quadrupole time-flight mass spectrometer, and the 31 kD target antigen was identified as U1-AsnRNP. To test the prevalence of anti-U1-AsnRNP antibody, sera from 93 patients including 36 squmaous carcinomas (SCC), 26 adenocarcinomas (Ad), and 31 small cell carcinomas (SCLC) were screened by Western blotting. The results demonstrated that anti-U1-A snRNP antibody was present in 50% of SCC sera, 26.9% of Ad sera and 54.8% of SCLC sera. In this paper, we report for the first time that anti-U1-AsnRNP antibody could be detected in the patients with lung cancer.     

Oncogene and tumor suppressor gene alterations in nasal tumors

The development of nasal tumors in humans and rodents is likely mediated through the accumulation of genetic alterations in genes that regulate cell proliferation, cell death and differentiation (oncogenes and tumor suppressor genes). By examination of the relationship between genetic alterations that are known to occur in human cancers with those induced in rodent tumors with defined carcinogenic exposures, biologically relevant mechanistic linkages of molecular events leading to tumors in rodents and humans can be established. Molecular genetic studies on nasal squamous cell carcinomas (SCC) in rats thus far have indicated the presence of oncogenes unrelated to the ras oncogene family and that p53 mutation occurs at a high frequency among the nasal SCC examined. The finding of p53 mutations in rat nasal SCC and the high prevalence of p53 mutations among human SCC, indicates that a common molecular alteration is shared between rodent and human SCC.     

Tumor markers in patients with chronic renal failure.

In order to evaluate the specificity of tumor markers in chronic renal failure, we have determined serum levels of carcinoembryonic antigen (CEA), carbohydrate antigen 19.9 (CA 19.9), carbohydrate antigen 50 (CA 50), alphafetoprotein (AFP), neuron-specific enolase (NSE), prostatic acid phosphatase (PAP), prostatic specific antigen (PSA), squamous cell carcinoma antigen (SCC), carbohydrate antigen 15.3 (CA 15.3) and carbohydrate antigen 125 (CA 125) in 30 patients with chronic renal failure and in 36 hemodialyzed patients without clinical evidence of neoplasia. CEA, CA 50, NSE and SCC frequently show increased serum levels, suggesting a renal metabolism, while others remain, generally, within the normal levels.     

The distribution of interleukin-19 in healthy and neoplastic tissue

The influence of interleukin (IL)-19, a recently discovered cytokine in the IL-10 family, on tissue is still unclear. Our aim was to determine the distribution of IL-19 expression and to delineate the cell types that express IL-19 in healthy and neoplastic tissue, because this information will significantly facilitate the exploration of its pathophysiological functions. We used tissue microarray technology and an immunohistochemical survey with an anti-IL-19 monoclonal antibody to examine the expression of IL-19 in 28 healthy and 15 neoplastic tissues. IL-19 protein was positively stained in 15 healthy tissue types and three major cell types: epithelial cells, endothelial cells, and macrophages. We also found that several types of tumor cells were positively stained for IL-19, especially in squamous cell carcinoma (SCC) of the skin, tongue, esophagus, and lung. SCC of the oral cavity expressed IL-19 mRNA and its receptors. In two cell lines derived from SCC of oral cavity tumor tissue, IL-19 specifically activated an intracellular signal and induced proliferation of the cells, which indicated that IL-19 may act in an autocrine manner in SCC tumors. This study provides important references for further investigation of the biological functions and clinical implications of IL-19 in humans.     

Small cell carcinoma of the bladder. Two cases diagnosed by urinary cytology

BACKGROUND: Primary small cell carcinoma (SCC) of the bladder is a rare but important entity. We report two cases of SCC of the bladder diagnosed by urinary cytology. CASES: A 71-year-old male (case 1) and a 79-year-old female (case 2) presented with asymptomatic gross hematuria. Urinary cytology in case 1 showed the presence of a few undifferentiated malignant small cells and many transitional cell carcinoma (TCC) cells with a bloody and necrotic background. The former cells were small and round, with naked, hyperchromatic nuclei and finely granular chromatin. Pathologic diagnosis after total cystectomy was TCC > SCC > adenocarcinoma, T2M0N0. Urinary cytology of case 2 showed the presence of many undifferentiated malignant small cells and many TCC cells with or without squamous metaplasia. Cytologic features of the former cells were almost the same as those in case 1. Moreover, these cells were neuroendocrine marker positive by immunocytochemistry. Pathologic diagnosis after tumor resection was SCC and TCC > squamous cell carcinoma, T1b. CONCLUSION: The prognosis of primary SCC of the bladder is usually poor. Because our cases were found by urinary cytology at a relatively early stage, both have been well, without any evidence of recurrence, 30 and 25 months after surgery even without adjuvant therapy.