The contribution of image cytometry to the characterization of clinical subgroups of lipomas

The aim of the present study was to investigate whether biological features determined through image cytometry are able to characterize clinical subpopulations of lipomas. Forty lipomas excised from 36 patients were studied. On the one hand, the tumors were clinically characterized by means of patient-related and pre- and post-operative features. On the other, the tumors were analyzed by means of the computer-assisted microscopy analysis of Feulgen-stained nuclei. This analysis generated 3 groups of biological quantitative variables describing morphonuclear aspects (i.e. the chromatin pattern of the cell nuclei), the nuclear DNA content (DNA ploidy level), and architectural features (such as the cell density and the topographical cell nuclei organization). Possible relations between the clinical and the biological features of the lipomas were investigated by means of univariate and multivariate statistical analyses. The results show the existence of such relations, in particular between the morphonuclear and architectural features of lipomas, on the one hand, and their consistency, volume and weight, on the other. Furthermore, multivariate analysis made it possible to distinguish two subpopulations of lipomas exhibiting different biological characteristics in terms of morphonuclear patterns.     

An immunohistochemical study of papillary tumors in the central nervous system

An immunohistochemical study was performed on the paraffin sections of 25 tumors in the central nervous system (CNS) with prominent papillary configurations. These tumors included seven metastatic papillary carcinomas, six choroid plexus tumors, four papillary ependymomas, two myxopapillary ependymomas, two papillary pituitary adenomas, two astroblastomas, one papillary meningioma, and one anaplastic astrocytoma with significant papillary changes. The panel of antibodies applied included anti-glial fibrillary acidic protein (GFAP), anti-carcinoembryonic antigen (CEA), anti-vimentin (VM), anti-S-100 protein (S-100 P), anti-cytokeratin, and anti-prealbumin antisera. All ependymomas and astroblastomas examined expressed both VM and GFAP, which were either absent or focally expressed in choroid plexus tumors. In contrast, all choroid plexus tumors showed anti-cytokeratin immunoreactivity that was absent in the ependymomas and astroblastomas. Five choroid plexus tumors also expressed S-100 P, thus differentiating them from metastatic carcinoma that showed negative immunostaining. Anti-CEA antisera immunoreactivity was seen in six metastatic tumors, whereas none of the primary CNS tumors expressed CEA. Prealbumin was expressed in four choroid plexus tumors and two metastatic tumors. Immunohistochemical typing using a panel of antibodies has allowed the differentiation of most of the papillary tumors in the CNS examined in this study.     

Histological spectrum of ependymomas and correlation of p53 and Ki-67 expression with ependymoma grade and subtype

BACKGROUND: Clinical and histological criteria for ependymoma prognosis are well recognized. Recently few studies have been done based on Immunohistochemistry for prognostication of these tumours. In this study we have correlated the histological spectrum with immmunoexpression of p53 and Ki67 in these tumors. AIMS: To know the incidence of ependymomas; study their morphological spectrum and to evaluate expression of P53 and Ki 67 in different morphological subtypes. MATERIAL AND METHOD: A retrospective study was preformed on 70 ependymomas received in a period between 1994 and 2001. Entire tissue received was processed for routine paraffin embedded H&E stained sections. Immunocytochemistry was performed using antibodies to GFAP, EMA, Pancytokeratin and synaptophysin, to differentiate papillary ependymoma from choroid plexus papilloma; clear cell ependymoma from oligodendroglioma and central neurocytoma; ependymoblastoma from other embryonal tumours. p53 and Ki-67 immunohistochemistry was performed to correlate their expression with various tumour grades and subtypes. RESULTS: There were 3 cases (4.2%) of Grade I ependymoma (2 cases of myxopapillary ependymoma and 1 case of subependymoma); 57 cases (81.5%) of ependymoma grade II (43 of these were of classical variety, 11 of clear cell ependymoma, 2 of papillary and 1 case of cellular ependymoma). There were 9 cases (12.8%) of anaplastic ependymoma (one of these was a clear cell ependymoma and 1 case (1.5%) of ependymoblastoma CONCLUSION: p53 and Ki67 indices can be used in routine diagnostic laboratories to supplement the tumor grade on histology and more studies with follow up should be performed to analyse the prognosis of different subtypes. The expression of Ki 67 and p53 was significantly higher in anaplastic ependymomas. 4 out of 11 cases of clear cell ependymomas showed higher Ki 67 indices as compared to classical grade II ependymomas, thus further highlighting the importance of differentiating the various subtypes.     

Image cytometry characterization of ploidy level, proliferative activity and chromatin pattern in 50 nasal polyps

A computer-assisted microscope analysis of Feulgen-stained nuclei was carried out on a series of 50 nasal polyps in order to try to identify specific biological subgroups. The present series of 50 nasal polyps includes single polyps both associated (n=9) and unassociated (n=9) with allergy and diffuse polyposis both associated (n=7) and unassociated (n=9) with allergy, cystic fibrosis (n=9) and ASA (aspirin-sinusitis-asthma) related polyposis (n=7). The computer-assisted microscope analysis provides 36 quantitative variables which include 1 variable describing proliferative activity, 9 describing the nuclear desoxyribonucleic acid distribution (DNA ploidy level) and 26 describing nucleus morphology, i.e. its size and chromatin pattern. The results show that the methodology proposed here enabled four major groups of nasal polyps to be identified, i.e. diffuse polyposis associated with allergy, cystic fibrosis-related polyposis, single polyps both associated and unassociated either with allergy and a fourth group including diffuse polyposis not associated with allergy and ASA-related polyposis. These four groups of nasal polyps differed markedly in their morphonuclear characteristics, but not in the proliferative activity- and DNA ploidy-related variables.     

DNA ploidy analysis and cell proliferation in congenital sacrococcygeal teratomas

BACKGROUND: Congenital sacrococcygeal teratoma is the most common germ cell tumor in infants and children. It usually is diagnosed at birth, is benign, and consists of fully differentiated mature tissues. Congenital sacrococcygeal teratomas (SCTs) also may contain immature tissues, most commonly of neural origin. The proportion of malignant teratomas increases with advancing age, but the relation between mature and immature SCTs is not well understood. Thus, it is very important to determine proliferative activity, DNA ploidy, and DNA index to predict biologic behavior of these tumors. METHODS: DNA ploidy and cell proliferation were analyzed by flow cytometry, and the expression of proliferating cell nuclear antigen (PCNA) and Ki-67 were analyzed immunohistochemically on paraffin embedded tissue. RESULTS: All the tumors that were surgically treated within 3 months after birth, including immature teratoma, were diploid. Strongly positive PCNA immunostaining was found in both immature teratomas, and weakly positive PCNA was found in nine cases. Weak positivity for Ki-67 was observed in 2 cases, and moderate positivity was observed in 6 cases including immature teratomas. CONCLUSION: The value of flow cytometry in the prediction of biologic behavior of congenital SCT should be analyzed further. Our results suggest that Ki-67 and especially PCNA may reflect the proliferative activity of these tumors.     

Prognostic value of Ki-67 (MIB-1) and p53 in ependymomas

To determine whether Ki-67 (MIB-1) and p53 have prognostic value in ependymomas, clinicopathologic study was undertaken in 29 patients with this tumor. The clinical course correlated well with the histological grade according to the World Health Organization (WHO) grading system, and it was the worst in patients with anaplastic ependymoma. The percent expression of MIB-1 and p53 correlated with the histological grade of malignancy. With regard to the subtypes of benign ependymoma, the clinical course was the worst in clear-cell ependymoma, which had a significantly higher expression of MIB-1 and p53 than the other subtypes. Tanycytic ependymoma showed the most benign clinical course and the lowest expression of MIB-1 and p53. Although the WHO grading generally tended to correlate with the clinical course of ependymomas, these two subtypes-clear-cell ependymoma and tanycytic ependymoma-exhibited biological properties different from those of other grade II ependymomas.     

In situ evaluation of growth fraction determined by monoclonal antibody Ki-67 and ploidy in surgically resected non-small cell lung cancers

Ploidy and growth fraction were analyzed by means of a computer-assisted image processor in surgically resected non-small cell lung cancer (NSCLC). This study was done in order (a) to evaluate the distribution of anti-Ki-67 immunostaining and (b) to correlate this distribution to ploidy status and pTNM stage of NSCLC. Thirty-two patients underwent a surgical resection for primary NSCLC following complete staging. Indirect immunoperoxidase reactions of monoclonal antibody Ki-67 were done on frozen tissue sections. Integrated optical density and index of stained nuclear surface were calculated by means of a computer-assisted image processor in 120 fields of each preparation in order to quantify the Ki-67 immunostaining. DNA content was determined by means of cytometry of Feulgen-stained cytological prints. The ploidy status was defined for each tumor by DNA index, percentage of hypodiploid cells, and type of DNA content histogram (near diploid, hyperdiploid, hypodiploid, and multiploid). Reproducibility of immunostaining quantitative analysis was demonstrated by iterative measurements of the same slide. Intratumoral heterogeneity of Ki-67 immunostaining induced integrated optical density variation assessed on six nonconsecutive tissue sections from at least two regions of the same tumor. This intratumoral variability was 15 times lower than integrated optical density variability between tumors. The Ki-67 immunostaining varied significantly according to the DNA content histogram type (P less than 0.05, Kruskal-Wallis test); most of the specimens with high Ki-67 immunostaining were multiploid or hypodiploid. Moreover, Ki-67 immunostaining correlated to the percentage of hypodiploid cells. Ki-67 immunostaining and ploidy status did not vary significantly according to the tumor-nodes-metastasis stage. We conclude that (a) quantitative analysis of Ki-67 immunostaining is a reliable evaluation of growth fraction in NSCLC if a large number of fields are analyzed to take into account intratumoral variability, (b) hypodiploidy and multiploidy are frequent abnormalities of DNA content, (c) Ki-67 immunostaining is significantly higher in hypodiploid and multiploid tumors. Thus, determination of growth fraction and ploidy in surgically resected NSCLC specimens may be considered as complementary prognostic parameters independent of the stage of the disease.     

乳腺肿瘤中DNA含量、Ki-67指数、肿瘤微血管密度（MVD）的定量研究

 目的 探讨DNA含量、Ki-67指数、肿瘤微血管密度（MVD）在乳腺肿瘤中意义、相关关系及计算机图象分析技术在肿瘤研究中的应用。方法 运用计算机图像分析系统对60例乳腺癌、28例乳腺癌前病变、25例乳腺良性病变中Ki-67、MVD及DNA含量进行定量检测。结果 Ki-67标记指数、微血管密度、细胞DNA相对倍体均值（U值）按乳腺良性病变、乳腺癌前病变及乳腺癌的顺序呈递增趋势，且随乳腺癌分级的增高而显著增高。结论 DNA相对倍体均值（U值）和Ki-67标记指数均能较好地反映肿瘤细胞增生活性。肿瘤MVD反映血管生成，是恶性实体性肿瘤生长和增殖的基础，是淋巴结转移的重要因素。计算机图像分析仪能够对肿瘤的组织切片进行多种参数的自动测量分析，为乳腺癌的自动化诊断及淋巴结转移判断提供方便。它将逐步应用到临床病理诊断工作中。     

Flow cytometric analysis of Ki-67 in invasive ductal carcinoma of the breast: correlation with tumor and patient characteristics.

Of all markers associated with cellular proliferation in breast carcinoma, Ki-67 has more often been correlated with prognosis in patients with these tumors than others. To investigate the relevance of Ki-67 determination at each phase of the cell cycle in the biological assessment of mammary carcinoma we applied bivariate Ki-67/DNA content analysis on samples from 154 resected primary lesions. Three Ki-67-derived indices including an overall and G1 and S+G2M indices were generated. These values were correlated with similar indices derived from flow cytometric DNA/RNA analysis and traditional clinicopathologic factors. The results show that overall Ki-67 indices do not correlate with flow cytometric values and clinicopathologic factors. Flow-derived Ki-67 and DNA S+G2M indices were positively correlated (p<0.0001, r=0.58). High indices for the S+G2M phase derived by both Ki-67 and DNA analysis were significantly correlated with DNA aneuploidy, high tumor grade, and negative hormonal status. We conclude that the proliferative fraction (S+G2M) by either Ki-67 or DNA analyses offers more practical and clinically relevant information in assessing the proliferative activity in mammary carcinoma.     

Evaluation of cell proliferation in breast carcinoma. Comparison of Ki-67 immunohistochemical study, DNA flow cytometric analysis, and mitotic count

Growth kinetics of 102 breast carcinomas were studied by immunohistochemical analysis with the monoclonal antibody Ki-67, which reacts with a nuclear antigen in proliferating cells. The results were correlated with ploidy and S-phase fraction (SPF) analyzed by DNA flow cytometric study and with mitotic count analyzed by light microscopic study. The proportion of Ki-67-positive cells (Ki-67 score) in breast carcinomas varied from 0.6% to 80% (median, 6.3%). Ki-67 scores were significantly higher in the DNA aneuploid than in the DNA diploid tumors. Ki-67 scores correlated significantly with tumor SPF in DNA aneuploid tumors. In DNA diploid tumors SPF showed no correlation with Ki-67 score. High Ki-67 scores were associated with high mitotic counts (P less than 0.0001) and histologic grade (P less than 0.0001). Nuclear pleomorphism, tubule formation, or lymph node status were not correlated with Ki-67 score. In conclusion, Ki-67 immunostaining correlates with other measures of cell proliferation (SPF, mitotic count) supporting its clinical significance.     

Supratentorial extraventricular ependymal neoplasms: a clinicopathologic study of 32 patients

BACKGROUND: Published research on the clinicopathologic features of extraventricular ependymal neoplasms of the cerebral hemispheres has been scant. METHODS: Thirty-two archival cases were studied to investigate the prognostic impact of clinicopathologic parameters including flow cytometry, the proliferation (Ki-67) labeling index, and p53 expression. RESULTS: Among these 32 cases were 2 subependymomas, 19 ependymomas, and 11 anaplastic ependymomas. No significant gender predilection was observed, and 45% of patients were in their second or third decade of life. The left cerebral hemisphere was 1.5 times more commonly involved. On available imaging studies, lesions were often cystic, with or without a mural nodule. Tumors expressed glial fibrillary acidic protein (87%), S-100 protein (77%), cytokeratin (43%), and epithelial membrane antigen (17%). Ki-67 proliferation index paralleled tumor grade. Immunoreactivity for p53 protein was observed in the 2 cases of subependymoma, in 10 of 11 anaplastic ependymomas, and in 6 of 17 ependymomas. Flow cytometry performed in 27 tumors revealed diploidy in 20 cases and aneuploidy in 4 cases (3 anaplastic and 1 classic ependymomas), with S-phase fraction ranging from 0.2-9.7. Eleven subjects were additionally treated with radiotherapy, and 3 with chemotherapy. Follow up was available in 25 (78%) patients. CONCLUSIONS: The results of the current study suggest that there is no significant relation between histopathology, Ki-67 proliferation index, p53 immunolabeling, tumor ploidy, and biologic behavior.     

Absence of simian virus 40 in human brain tumors from northern India

Simian virus 40 (SV40), a monkey polyomavirus, was a contaminant of early poliovirus vaccines administered to millions of individuals in the 1950s and early 1960s. SV40 causes brain tumors in laboratory animals, and SV40 DNA sequences have been variably identified in human choroid plexus tumors and ependymomas. We studied the possible association between SV40 and human brain tumors in northern India, where humans have frequent contact with SV40-infected rhesus macaques. DNA from pathologic specimens from 33 ependymomas, 14 choroid plexus tumors and 18 control brain tissues (contused brain, brain metastases) was extracted and analyzed under masked conditions. We used real-time PCR to detect and quantify SV40 (T antigen) and human (GAPDH) DNA sequences. The SV40 PCR assay detected as few as 10 copies of SV40 DNA and had a linear range from 1 x 10(2) to 1 x 10(6) copies. SV40 DNA was detected in 1 specimen (an ependymoma). However, few SV40 DNA copies were detected in this sample (<10 copies, equivalent to <1 copy/350 cells, based on simultaneous GAPDH quantification), and SV40 was not detected when this sample was retested. Our findings do not support a role for SV40 in choroid plexus tumors or ependymomas from northern India.     

Fas Engagement Increases Expression of Interleukin-6 in Human Glioma Cells

The aim of this investigation was recognition of the epithelial differentiation and proliferative activity of ependymomas in the spinal cord compared with astrocytomas in the spinal cord and ependymoma in the brain. We investigated histopathological and immunohistochemical examination in thirty-five cases, including eleven ependymomas, thirteen astrocytomas in the spinal region and eleven ependymomas in the intracranial region. An anti-epithelial membrane antigen antibody (EMA), and two types of anti-cytokeratin antibodies, CAM 5.2 (45 and 52kDa) and keratin (56 and 64kDa) were applied as epithelial markers. The proliferative potential of the tumors was investigated using the Ki-67 labeling index (LI, %). Histologically, perivascular pseudorosettes were seen in all of the spinal and intracranial ependymomas. There were few ependymal structures in the spinal ependymomas except in the myxopapillary type. Immunohistochemical study demonstrated that nine (82%) were immunoreactive for EMA, eight (73%) were immunoreactive for CAM 5.2 and three (27%) were immunoreactive for keratin in the spinal ependymomas. In the spinal astrocytomas, no tumors were immunoreactive for EMA or CAM 5.2. One of thirteen cases was immunoreactive for keratin. The Ki-67 LI of the spinal ependymomas was lower than that of the intracranial ependymoma (p < 0.01). Epithelial differentiation was found in the ependymomas, which may reflect the differences in histological and biological features between ependymomas and astrocytomas in the spinal cord. Regional differences in ependymomas may influence not only clinical features but also histo-pathological characteristics.     

Growth fractions in gastric carcinomas determined with monoclonal antibody Ki-67

The growth fractions of 101 gastric carcinomas were determined in situ by immunostaining with the monoclonal antibody Ki-67 and the results correlated with the histopathologic findings, bromodeoxyuridine (BrdU) labeling index, and DNA ploidy pattern. DNA ploidy patterns were determined by flow cytometric analysis. The Ki-67 labeling rates are rated from 4.6% to 82% (mean, 22%). A significant correlation was found between Ki-67 labeling rates and BrdU labeling indices. Sixty-seven percent of tumors with lymph node metastases showed Ki-67 labeling rates of less than 22%, whereas 33% of tumors without lymph node metastases showed Ki-67 labeling rates of less than 22%. There was a significant correlation between these two groups. Tumors with vessel invasion more often have higher Ki-67 labeling rates than those without vessel invasion. By the DNA ploidy classification, the mean Ki-67 labeling rates of aneuploid tumors was significantly higher than that of diploid tumors. This method yielded similar results to those obtained by BrdU labeling and flow cytometric study. The measurement of Ki-67 labeling rates may be useful to decide the therapeutic modalities.     

Immunohistochemical Prognostic Markers in Intracranial Ependymomas: Systematic Review and Meta-Analysis

Distinction between grade II ependymomas and anaplastic ependymomas based on histopathological examination solely is problematic and, therefore, the management of intracranial ependymomas remains controversial. The aim of this study was to conduct a systematic review (SR) and meta-analysis (MA) of data published on immunohistochemical prognostic markers (IPM) in intracranial ependymomas (IE), and to establish an evidence-based perspective on their clinical value. Following the extensive search based on a strictly defined group of key words, 30 studies reporting results on IPM in IE were identified. Due to a pronounced inter-study heterogeneity, only 14 publications fulfilled the criteria for inclusion into SR. From the total of 67 immunohistochemical markers, 18 were found to correlate with prognosis. However, owing to inadequate data publishing, MA could be performed only with data on proliferation marker MIB-1 (Ki-67) from 5 publications, including 337 patients: The pooled hazard ratio for overall survival was 3.16 (95% confidence interval = 1.96–5.09; p < 0.001) implicating that patients suffering from tumors with higher immunohistochemical expression of MIB-1 had a significantly worse outcome. Marked inter-study heterogeneity and incomplete data publishing in primary studies significantly limited extent of the SR, and the possibility of performing MA. Although the prognostic impact of MIB-1 immunoexpression in IE could be confirmed, there remains lack of further reliable IPM that could be used in routine diagnosis. We encourage to search for new, useful markers, as well as to standardize lab-techniques and data interpretation algorithms across laboratories in order to increase data compatibility.     

<Superscript>99m</Superscript>Tc-Tetrofosmin brain SPECT in the assessment of meningiomas—correlation with histological grade and proliferation index

Meningiomas account for about 30% of all intracranial tumors. Evaluation of their proliferation rate is useful for assessing their biological behavior. We evaluated prospectively whether (99m)Tc-Tetrofosmin ((99m)Tc-TF) uptake in meningiomas correlates with cellular proliferative activity and with tumor grade. We prospectively studied 18 meningioma cases. Brain single-photon emission computed tomography (SPECT) by (99m)Tc-TF was performed within a week prior to surgical excision. In the excised tumor specimens we assessed Ki-67 antigen expression. 14 of 18 patients had benign meningiomas, while the remaining four had anaplastic meningiomas. A significant correlation was found between both (99m)Tc-TF uptake and tumor grade (r = 0.722, P = 0.001) and between (99m)Tc-TF uptake and Ki-67 expression (r = 0.930, P < 0.001). There was a significant correlation between the intensity of tracer uptake and tumor recurrence at 1 year postoperative (r = 0.574, P = 0.02). This pilot study implies that (99m)Tc-TF brain SPECT could prove useful in differentiating benign from anaplastic meningiomas and is a potential indicator of their proliferative activity.     

Hypodiploidy, Ki-67 growth fraction and prognosis of surgically resected lung cancers.

One hundred and thirty-seven lung cancer patients (123 non-small-cell lung cancers (NSCLC), 10 small-cell lung cancers (SCLC) and four carcinoid tumours) who underwent surgery in an attempt at complete resection were prospectively entered in a study whose aim was to determine the prognostic significance of a hypodiploidy or a multiploidy pattern of tumour cell DNA content and a high immunohistochemical reactivity of Ki-67, a nuclear antigen related to the cell cycle. Indirect immunoperoxidase reactivity of Ki-67 on frozen tumour tissue sections was evaluated both visually, using a classical semiquantitative scale, and by means of a computer-assisted image processor. Cell DNA content analysis was done using static computer-assisted cytometry on tumour cytological prints stained by the pararosaline Feulgen-Schiff technique. The ploidy was characterised for each tumour by DNA index (DI), percentage of hypodiploid cells and type of DNA content histogram (near diploid, hyperdiploid, hypodiploid and multiploid). Ki-67 immunostaining was negative in 64 tumours (48%) and positive in 69 (52%). DNA histogram classification disclosed 57 (42%) near diploid tumours. Among the 80 (58%) aneuploid tumours, 16 were hypodiploid, 44 hyperdiploid and 20 multiploid. The prevalence of both a positive Ki-67 immunostaining and an aneuploid DNA histogram differed according to histology as SCLC demonstrated a higher frequency of both features when compared with NSCLC and carcinoid tumours. On the other hand, Ki-67 immunostaining and ploidy did not significantly differ according to degree of differentiation, nodal status and Mountain''s stage grouping. The percentage of cells in the hypodiploid modal DNA was significantly higher for tumours which demonstrated a high Ki-67 immunostaining, suggesting a link between growth fraction and DNA content abnormalities. In univariate analysis, survival did not differ significantly according to either the Ki-67 immunohistochemical reactivity or the DNA index. Patients with a hypodiploid tumour had a shorter survival than patients with other DNA histogram patterns but, owing to the low frequency of hypodiploidy, this difference did not reach statistical significance. In Cox''s proportional hazard model, an SCLC histology, an advanced tumour status, a positive nodal status and a hypodiploid tumour (hazard ratio: 2.070; 95% confidence interval 1.041-4.116) were significant determinants of survival. We conclude that hypodiploidy in lung cancer is a distinct DNA content abnormality as it contributes significantly to prognosis. Neither visually assessed nor computer-generated Ki-67 immunostaining measurements significantly determine prognosis.     

P16/Ki-67双染联合DNA倍体分析在ASCUS分流诊断中的应用价值

目的： 探讨P16/Ki-67双染联合DNA倍体分析在宫颈意义不明的不典型鳞状细胞（ASCUS）分流诊断中的应用价值。方法： 选取2016年12月-2018年8月上海市长宁区妇幼保健院收治的经液基细胞学（LCT）诊断为ASCUS的患者115例，同时行P16/Ki-67双染检测，DNA倍体分析，且以阴道镜病理活检结果为金标准。结果： 115例患者阴道镜病理活检结果提示，宫颈上皮瘤变2级及以上（CIN2+）45例，其中CIN2级20例，CIN3级23例，鳞癌2例；P16/Ki-67双染检测CIN2+的灵敏性、特异性、诊断符合率分别为77.8%、77.1%、77.4%；DNA倍体分析检测CIN2+的灵敏性、特异性、诊断符合率分别为71.1%、34.3%、48.7%，且DNA倍体阴性的细胞标本中P16/Ki-67双染检测结果均为阴性；DNA倍体分析联合P16/Ki-67双染检测CIN2+的灵敏性、特异性、诊断符合率分别为92.9%、71.4%、86.3%，其中诊断符合率明显高于单纯采用DNA倍体分析（P < 0.05）。结论： P16/Ki-67双染相较于DNA倍体分析检测CIN2+具有更高灵敏度和特异度，两者联合检测能有效提高准确率，可辅助用于ASCUS的分流诊断。     

P16/Ki-67双染联合DNA倍体分析在ASCUS分流诊断中的应用价值

目的： 探讨P16/Ki-67双染联合DNA倍体分析在宫颈意义不明的不典型鳞状细胞（ASCUS）分流诊断中的应用价值。方法： 选取2016年12月-2018年8月上海市长宁区妇幼保健院收治的经液基细胞学（LCT）诊断为ASCUS的患者115例，同时行P16/Ki-67双染检测，DNA倍体分析，且以阴道镜病理活检结果为金标准。结果： 115例患者阴道镜病理活检结果提示，宫颈上皮瘤变2级及以上（CIN2+）45例，其中CIN2级20例，CIN3级23例，鳞癌2例；P16/Ki-67双染检测CIN2+的灵敏性、特异性、诊断符合率分别为77.8%、77.1%、77.4%；DNA倍体分析检测CIN2+的灵敏性、特异性、诊断符合率分别为71.1%、34.3%、48.7%，且DNA倍体阴性的细胞标本中P16/Ki-67双染检测结果均为阴性；DNA倍体分析联合P16/Ki-67双染检测CIN2+的灵敏性、特异性、诊断符合率分别为92.9%、71.4%、86.3%，其中诊断符合率明显高于单纯采用DNA倍体分析（P < 0.05）。结论： P16/Ki-67双染相较于DNA倍体分析检测CIN2+具有更高灵敏度和特异度，两者联合检测能有效提高准确率，可辅助用于ASCUS的分流诊断。     

Predictive value of Ki-67, p53 protein, and DNA content in the diagnosis of gastric carcinoma.

BACKGROUND: The ability to make a precise preoperative diagnosis is a valuable and effective method in improving the prognosis of patients with gastric carcinoma. The authors examined retrospectively whether preoperative histopathologic analysis with p53 protein, Ki-67 labeling index, and DNA ploidy along with preoperative radiographic and endoscopic findings led to a precise preoperative diagnosis of patients with gastric carcinoma. METHODS: Histopathologic analysis of p53 protein, Ki-67 labeling index, and DNA content was performed on formalin fixed, paraffin embedded tissue. Tissue sections from endoscopic and surgically resected specimens were stained immunohistochemically for p53 protein and Ki-67 labeling index, and the cell nuclear DNA content of the surgically resected primary lesion was measured using a microspectrophotometer. These analyses were performed on 16 patients with early gastric carcinoma (EGC) who were diagnosed with advanced gastric carcinoma (AGC) based on the preoperative imaging findings and on 15 patients with AGC who were diagnosed preoperatively with EGC. RESULTS: Overexpression of p53 in the AGC group was significantly more frequent compared with that in the EGC group (P = 0.0386). With regard to the correlation between lymph node metastases and p53 overexpression, there was no apparent relation in either the AGC group (P = 0.648) or the EGC group (P = 0.726). The AGC group had significantly higher Ki-67 labeling indices compared with the EGC group (P = 0.0195). There was complete concordance between endoscopic and surgically resected specimens with regard to the p53 and Ki-67 labeling index findings. DNA ploidy in the primary tumor did not differ between the AGC and EGC groups. The survival rates for the EGC group were significantly superior to those for the AGC group (P = 0.0312). CONCLUSIONS: The findings of the current study suggest that in routine clinical practice, the combination of preoperative imaging findings in addition to Ki-67 labeling indexes, and p53 protein analyses may be useful for the accurate diagnosis of EGC; however, DNA ploidy did not appear to reflect the growth potential of gastric carcinoma.