Ploidy and fruit trait variation in oil-tea Camellia:Implications for ploidy breeding

Plant polyploidy often occurs in conjunction with higher yield and superior quality.Therefore,obtaining polyploid germplasms is a significant part of breeding.The oil-tea Camellia tree is an important native woody plant that produces high-quality edible oil and includes many species of Camellia with different ploidies.However,whether higher ploidy levels in oil-tea Camellia trees are related to better traits remains unclear.In this study,the ploidy levels of 30 different oil-tea Camellia strains in three different species in the Sect.Paracamellia were determined by flow cytometry and chromosome preparation,and the phenotypic characteristics and fatty acid compositions of the fruits were examined by field observations and laboratory analyses.The correlations between the ploidy level of oil-tea Camellia and the main traits of the fruit were investigated.Our results showed that 10 Camellia lanceoleosa strains were diploid,10 Camellia meiocarpa strains were tetraploid and 10 Camellia oleifera strains were hexaploid.Hexaploid C.oleifera had larger fruit size and weight,more seeds per fruit,greater seed weight per fruit,higher oil content and greater yield per crown width than tetraploid C.meiocarpa and diploid C.lanceoleosa,but their fruit peel thickness and fresh seed rate were significantly lower,and these traits were significantly correlated with ploidy level.In addition,in terms of fatty acid composition,hexaploid C.oleifera had a higher oleic acid content than tetraploid C.meiocarpa and diploid C.lanceoleosa,but their linoleic acid,linolenic acid and arachidonic acid contents were lower.The contents of palmitic acid,stearic acid and total unsaturated fatty acids were not significantly correlated with ploidy level.In conclusion,certain correlations exist between the main characteristics of oil-tea Camellia fruit and the ploidy level,and increasing the ploidy level led to an increase in fruit yield with no effect on oil composition.The discovery of variations in the main characteristics of oil-tea Camellia fruit with different ploidies will facilitate germplasm innovation and lay a foundation for ploidy breeding and mechanistic research on fruit traits.

The prognostic molecular markers in hepatocellular carcinoma

The prognosis of hepatocellular carcinoma (HCC) still remains dismal, although many advances in its clinical study have been made. It is important for tumor control to identify the factors that predispose patients to death. With new discoveries in cancer biology, the pathological and biological prognostic factors of HCC have been studied quite extensively. Analyzing molecular markers (biomarkers) with prognostic significance is a complementary method. A large number of molecular factors have been shown to associate with the invasiveness of HCC, and have potential prognostic significance. One important aspect is the analysis of molecular markers for the cellular malignancy phenotype. These include alterations in DNA ploidy, cellular proliferation markers (PCNA, Ki-67, Mcm2, MIB1, MIA, and CSE1L/CAS protein), nuclear morphology, the p53 gene and its related molecule MD M2, other cell cycle regulators (cyclin A, cyclin D, cyclin E, cdc2, p27, p73), oncogenes and their receptors (such as ras, c-myc, c-fms, HGF, c-met, and erb-B receptor family members), apoptosis related factors (Fas and FasL), as well as telomerase activity. Another important aspect is the analysis of molecular markers involved in the process of cancer invasion and metastasis. Adhesion molecules (E-cadherin, catenins, serum intercellular adhesion molecule-1, CD44 variants), proteinases involved in the degradation of extracellular matrix (MMP-2, MMP-9, uPA, uPAR, PAI), as well as other molecules have been regarded as biomarkers for the malignant phenotype of HCC, and are related to prognosis and therapeutic outcomes. Tumor angiogenesis is critical to both the growth and metastasis of cancers including HCC, and has drawn much attention in recent years. Many angiogenesis-related markers, such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived endothelial cell growth factor (PD-ECGF), thrombospondin (TSP), angiogenin, pleiotrophin, and endostatin (ES) levels, as well as intratumor microvessel density (MVD) have been evaluated and found to be of prognostic significance. Body fluid (particularly blood and urinary) testing for biomarkers is easily accessible and useful in clinical patients. The prognostic significance of circulating DNA in plasma or serum, and its genetic alterations in HCC are other important trends. More attention should be paid to these two areas in future. As the progress of the human genome project advances, so does a clearer understanding of tumor biology, and more and more new prognostic markers with high sensitivity and specificity will be found and used in clinical assays. However, the combination of some items, i.e., the pathological features and some biomarkers mentioned above, seems to be more practical for now.

DNA ploidy and c-Kitmutation in gastrointestinal stromal tumors

AIM: To investigate the prognostic significance of c-Kitgen emutation and DNA ploidy in gastointestinal stromal tumors (GISTs).METHODS: A total of 55 cases of GISTs were studied for the expression of c-Kit by immunohistochemistry, and the c-Kit gene mutations in exons 9, 11, 13, and 17 were detected by polymerase chain reaction-single strand confirmation polymarphism (PCR-SSCP) and denaturing high performance liquid chromatography (D-HPLC) techniques. DNA ploidy was determined by flow cytometry.RESULTS: Of the 55 cases of GISTs, 53 cases (96.4%) expressed c-Kit protein. The c-Kit gene mutations of exons 11 and 9 were found in 30 (54.5%) and 7 cases (12.7%),respectively. No mutations were found in exons 13 and 17.DNA aneuploidy was seen in 10 cases (18.2%). The c-Kit mutation positive GISTs were larger in size than the negative GISTs. The aneuploidy tumors were statistically associated with large size, high mitotic counts, high risk groups, high cellularity and severe nuclear atypia, and epithelioid type.There was a tendency that c-Kit mutations were more frequently found in aneuploidy GISTs.CONCLUSION: DNA aneuploidy and c-Kit mutations can be considered as prognostic factors in GISTs.

DNA倍体分析在食管良恶性疾病转归及预后研究中的应用

DNA非整倍体的出现是恶性肿瘤的特征性标志之一。食管上皮细胞的不典型增生中,DNA异倍体的出现是恶变的早期信号。采用流式细胞术检测癌细胞生物学特性,是从分子生物学水平探索肿瘤生物学特性及其发展规律。测量和分析细胞核DNA含量与倍体对恶性肿瘤的病理诊断、恶性程度判定、疗效估价、预测预后具有重要价值。本文主要综述了肿瘤细胞DNA倍体分析及其临床应用的现状,希望对其应用于临床以提高对食道良恶性疾病诊断的准确性和预测食道癌前病变的发展转归起到积极的作用。

DNA ploidy analysis and expression of MMP-9, TIMP-2, and E-cadherin in gastric carcinoma

AIM： To investigate DNA ploidy and expression of MMP-9, TIMP-2, and E-cadherin in gastric carcinoma and to explore the mechanism of invasion and metastasis of gastric carcinoma. METHODS： Immunohistochemical methods were used to detect the expressions of MMP-9, TIMP-2, and E-cadherin in 156 cases, including 99 cases of gastric carcinoma, 16 cases of adjacent noncancerous mucosa, 16 cases of distant metastases and 25 cases of metastatic lymph node （LN） from gastric carcinoma. Flow cytometry DNA ploidy and S-phase fraction （SPF） analysis were performed on 57 cases, including 47 cases of gastric cancer, 6 cases of adjacent noncancerous mucosa, and 4 cases of distant metastatic cancer. RESULTS： The expression of MMP-9 was significantly correlated with Lauren＇s classification, Borrmann＇s classification, LN metastasis, tumor metastasis, and TNM stage, as well as depth of invasion （all P〈0.05）. The positive rate was lower in noncarcinoma than in carcinoma （31.3% vs 66.7%, P〈0.01）. The expression of TIMP-2 was significantly correlated with Borrmann＇s classification, LN metastasis, and the depth of invasion （all P〈0.05）. The expression of E-cadherin was significantly correlated with differentiation, Lauren＇s classification, Borrmann＇s classification, and LN metastasis, as well as the depth of invasion （P〈0.01 or P〈0.05）. E-cadherin was less expressed in carcinoma than in noncarcinoma （42.4% vs 87.5%, P〈0.01）. There was a positive correlation between MMP-9and TIMP-2 and a negative correlation between MMP-9 and E-cadherin, but no correlation between TIMP-2 and E-cadherin. Also there was a positive correlation between DNA aneuploid rate and differentiation and LN metastasis. SPF that was higher than 15% was positively correlated with tumor size, differentiation and LN metastasis. And there was a significant difference between carcinoma and noncarcinoma in DNA aneuploid rate and SPF. CONCLUSION： With tumor progression and development of heterogeneity, the abnormal expressions of MMP-9, TIMP-2, and E-cadherin or DNA aneuploid rate or high SPF gradually increases, suggesting that they play a crucial role in gastric carcinoma progression. 2005 The WJG Press and Elsevier Inc. All rights reserved

Clinical application of DNA ploidy to cervical cancer screening: A review

Screening for cervical cancer with DNA ploidy assessment by automated quantitative image cytometry has spread throughout China over the past decade and now an estimated 1 million tests per year are done there. Compared to conventional liquid based cytology, DNA ploidy has competitive accuracy with much higher throughput per technician. DNA ploidy has the enormous advantage that it is an objective technology that can be taught in typically 2 or 3 wk, unlike qualitative cytology, and so it can enable screening in places that lack sufficient qualified cytotechnologists and cytopathologists for conventional cytology. Most papers on experience with application of the technology to cervical cancer screening over the past decade were published in the Chinese language. This review aims to provide a consistent framework for analysis of screening data and to summarize some of the work published from 2005 to the end of 2013. Of particular interest are a few studies comparing DNA ploidy with testing for high risk human papilloma virus(hrH PV) which suggest that DNA ploidy is at least equivalent, easier and less expensive than hrH PV testing. There may also be patient management benefits to combining hr HPV testing with DNA ploidy. Some knowledge gaps are identified and some suggestions are made for future research directions.

Relationship between DNA ploidy, expression of ki-67 antigen and gastric cancer metastasis

AIM To evaluate the relationship between the expression of Ki 67 antigen and the pathobiological behaviours of gastric cancers especially their distant metastases. METHODS Fifty six specimens of gastric cancer routinely fixed in formalin and embedded in paraffin (FFEP) were studied by immunohistochemical method. RESULTS Expression of Ki 67 antigen was significantly related to the distant metastases to liver, ovary and adrenal gland ( P <0 005), but not related to the histological type, growth pattern, depth of invasion, histological differentiation and the metastases to local lymph nodes ( P >0 05). Furthermore, the Ki 67 antigen expression was significantly related to the DNA aneuploidy pattern, which is closely related to poor prognosis ( P <0 05). CONCLUSION Overexpression of Ki 67 can be used as an objective marker of the proliferative activity for predicting prognosis of gastric cancer and metastatic potential to distant organs.

Cytological Evaluation and Karyotype Analysis in Plant Germplasms of Elytrigia Desv.

Elytrigia Desv. is widely distributed throughout the world and is represented with species of various levels of ploidy including diploids, tetraploids, hexaploids, octaploids, and decaploids. The distribution pattern of these ploidy levels, however, is not well-defined. In this study, the levels of ploidy for 64 accessions of Elytrigia from 25 countries were determined with microscopic procedures. The results showed that accessions of E. intermedia and E. repens were grouped into three distinct levels of ploidy including diploids, tetraploids and hexaploids. For E. elongata, E. pontica, and E. caespitosa, it was found that two ploidy levels presented, and only one ploidy level was in those of E. hybrid, E. pycnantha, E. pungens, E. juncea, and E. alatavica. Karyotype analysis indicated that the karyotype formula of diploid, tetraploid and hexaploid of E. intermedia was 2n = 2x = 14 = 6m ＋ 6sm ＋ 2st, 2n = 4x = 28 = 2M ＋ 10m ＋ 16sm and 2n = 6x = 42 = 4M ＋ 18m ＋ 20sin, respectively. Furthermore, the karyotype formula of three germplasms in tetraploid of E. intermedia was 2n=4x=28 =2M＋ 10m＋ 16sm, 2n=4x=28=4M＋22m （sat）＋2sm and 2n=4x=28 =4M＋ 12m＋ 12sm （sat）, which were not completely uniform. Therefore, it could be suggested that the studies about chromosome constitution would be helpful for the detail understanding of the diversity of germplasm resource in Elytrigia and promoting the utilization in the crop molecular breeding.

Analysis of DNA Ploidy, Cell Cycle and Ki67 Antigen in Nasopharyngeal Carcinoma by Flow Cytometry

Summary: The expression of DNA ploidy, the cell cycle and Ki67 antigen in nasopharyngeal carcinoma (NPC) were studied and their relationship with the clinical biological behaviors and prognosis of NPC was evaluated. Biopsied specimens of NPC were made into cell suspension. By using cytometric double labeling Ki67 and DNA method, the expression of DNA ploidy, the cell cycle and Ki67 antigen were analyzed. The patients were followed-up for about 3 years and the relationship between the above-mentioned parameters and the clinical biological behavior and prognosis of NPC were evaluated. Of the 62 cases of NPC, the DNA aneuploid accounted for 29.03 %. The S phase cells accounted for 0 to 54 % in the cell cycle and the positive expression of Ki67 ranged from 0 to 52 %. There were 40 cases of LPI (64.5 %) including 15 negative cases and 22 cases of HPI (35 5 %) respectively. The DNA anueploid content was positively related to the S phase cells. The patients having a low expression of Ki67 or DNA aneuploid in tumor cells were not sensitive to chemotherapy, liable to metastasis to distant organs and had a poor prognosis, while Ki67 showed no correlation with DNA ploidy and the cell cycle. It was suggested that DNA ploidy and Ki67 could be used as an independent and objective marker to evaluate the radiosensitivity and prognosis of NPC.

Morphological characteristics of leaf epidermis and size variation of leaf,flower and fruit in different ploidy levels in Buddleja macrostachya (Buddlejaceae)

Buddleja macrostachya （Buddlejaceae） is a widespread shrub native to the Sino-Himalayan mountains and beyond. It has been found to occur at two ploidy levels, hexaploid, 2n=6x=114 and dodecaploid, 2n= 12x=228. To determine if morphological characters might be used as indicators of ploidy levels, we measured floral and fruit length, relative and absolute leaf size, trichome density on both leaf surfaces, and stomatal density and length in different populations orB. macrostachya. In general, flower and fruit length, absolute leaf size, and stomatal length in,eased with an increase at ploidy level （P〈0.01）, whereas adaxial cell and stomatal density decreased with an increase at ploidy level （P〈0.01）. We found no conspicuous differences in relative leaf size （P〉0.05） in different populations. Other characters studied such as trichome type, cuticular membrane and ornamentation of stomata, cell and stomatal shape, and anticlinal wall pattern were quite constant in this species. Thus it appears that flower and fruit length, absolute leaf size, and stomatal frequency and length can be used to distinguish hexaptoid from dodecaploid cytotypes either in the field or in herbarium specimens.

Physiological and molecular characteristics of two ploidy mutants in Myrica rubracv.Dongkui

In this study, two ploidy mutant lines ofMyrica rubra cv. Dongkui （DK） were identiifed and named as DB1 and DB2. The lforal organ, leaf cel structure, ploidy, and number of chromosomes of the two mutants were investigated. Meanwhile, anthocyanin contents at different developmental stages were analyzed, and the Cy-3-glu contents of DB1 and DB2 at the ful ripe stages are signiifcantly higher than that of DK by 27.84 and 23.51%, respectively. Furthermore, 6 RNA libraries at two developmental stages （young fruit stage and ful ripe stage） were built for RNA-Seq. By mapping to the reference database, 28407, 28043, and 28683 genes were detected in the young fruit of DB1, DB2, and DK, respectively, while 28040, 22256, and 27351 genes were detected in the ful ripe stage, respectively. There were 281 differentialy expressed genes between DB1 and DK, with 123 and 158 genes up-regulated and down-regulated, respectively, and 47 differentialy expressed genes between DB2 and DK, of which 8 and 39 genes were up-regulated and down-regulated. Using real-time PCR, the expression levels of the eight functional genes at different developmental stages of the fruit were also analyzed. These comprehensive analyses showed that both mutants are different from DK, which is the result of natural doubling of ploidy, thereby generating a pleiotropic effect. As we known, it is the ifrst report to study the relationship between bayberry ploidy alterations and genes involved in regulation of fruit mutations, which wil help to identify the morphological and cytological characteristics ofM. rubragermplasm, and provide a theoretical basis and technical support for genetic improvement and creation of breeding resources.

DNA PLOIDY，EXPRESSION OF p53 PROTEIN AND METASTATIC BEHAVIOUR OF GASTRIC CARCINOMA

DNA ploidy of 57 gastric carcinomas with metastases(12 liver,1 adrenal,4 ovary and 48 lymph node) were measured by flow cytometry.DNA anueploidy was significantly related to liver metastases:9 out of 12 gastric carcinomas with liver metastases were anueploid(75%) as compared to 13 out of 45(28.8%) of cases without liver metastases(P<0.01);the one gastric carcinoma with adrenal metastasis was also anueploid.DNA ploidy was not related to ovarian or lymph node metastases.Another interesting finding was that all of 3 gastric carcinomas with liver metastases which showed a diploid DNA pattern,expressed p53 protein, while all of 3 carcinomas with liver metastases but no p53 protein expression were anueploid.The expression of p53 protein was not related to ovarian metastases.The results suggested that an anueploid DNA pattern and the expression of p53 protein are both objective markers valuable in predicting high risk potential of metastases to the liver,and that the combined detection of these markers can be a most useful method in the follow-up of Patients with gastric carcinoma in detecting those at high risk of developing metastases following surgical resection.Also the poorer prognosis of Patients with gastric carcinoma showing an anueploid DNA pattern may be related to the development of distant organ metastases through the blood vascular system.Furthermore,the clone of gastric carcinoma cells which accumulate p53protein or show an anueploid DNA pattern may have a causative role in the development of liver(&.adrenal) metastases.

Study on the Relationship Between the Ploidy Level of Microspore-Derived Plants and the Number of Chloroplast in Stomatal Guard Cells in Brassica oleracea

The relationship between the ploidy level of microspore-derived plants and chloroplast number in stomatal guard cells was studied in cabbage, broccoli, and Chinese kale. In the experiment, distribution statistics analysis and t-test were used to perform statistical analysis on chloroplast number of different ploidy level in those stomatal guard cells mentioned above, and morphology identifying and chromosome counting were used to test accuracy of counting chloroplast number in stomatal guard cells. The chloroplast average number in stomatal guard cells was very similar among the different leaf positions on the same plant and among significantly among the different ploidy the different locations in the same stoma in the same variety. All the leaf, while the chloroplast number varied distributions of the chloroplast number in different ploidy stoma were normal distribution fitted. A correlation has been established between ploidy and chloroplast number in the stomatal guard cells. In every single stoma of microspore-derived plants, the chloroplast number for a haploid should not be more than 10, diploids 11 to 15, and polyploids more than 15. The accuracy of this method for identification of different ploidy plants was 93.93%. Furthermore, the accuracy of this method was reliable and did not vary with the plants growth conditions. Therefore, the chromosome ploidy of plants derived from microspore culture in cabbage, broccoli, and Chinese kale can be identified by simply counting the chloroplast number in stomatal guard cells.

Plant regeneration via protoplast electrofusion in cassava

Protoplast electrofusion between callus protoplasts of cultivar TMS60444 and mesophyll protoplasts of cultivar SC8 was performed as an approach for the genetic improvement of cassava.The fusion products were subsequently cultured in protoplast culture medium(TM2 G) with gradual dilution for approximately 1-2 months.Then the protoplast-derived compact calli were transferred to suspension culture medium(SH) for suspension culture.The cultured products developed successively into embryos,mature embryos,and shoots on somatic embryo emerging medium(MSN),embryo maturation medium(CMM),and shoot elongation medium(CEM),respectively.And the shoots were then rooted on Murashige and Skoog(1962) medium(MS).Sixty-six cell lines were obtained and 12 of them developed into plantlets.Based on assessment of ploidy level and chromosome counting,four of these plantlets were tetraploid and the remaining eight were diploid.Based on assessment of ploidy level and simple sequence repeat(SSR) analysis,nine tetraploid cell lines,one diploid variant plant line and nine variant cell lines were obtained.The diploid variant plant line and the nine variant cell lines all showed partial loss of genetic material compared to that of the parent TMS60444,based on SSR patterns.These results showed that some new germplasm of cassava were created.In this study,a protocol for protoplast electrofusion was developed and validated.Another important conclusion from this work is the confirmation of a viable protocol for the regeneration of plants from cassava protoplasts.Going forward,we hope to provide technical guidance for cassava tissue culture,and also provide some useful inspiration and reference for further genetic improvement of cassava.

Effect of ploidy level on expression of lycopene biosynthesis genes and accumulation of phytohormones during watermelon(Citrullus lanatus) fruit development and ripening

The difference between lycopene and phytohormone levels among diploid, triploid and tetraploid plants of two watermelon cultivars during fruit growth and ripening was studied. The expression pattern of five genes（phytoene synthase（PSY1）, phytoene desaturase（PDS）, ζ-carotene desaturase（ZDS）, carotenoid isomerase（CRTISO）, and lycopene β-cyclase（LCYB）） was analyzed in details. In red-fleshed cultivar Mimei, lycopene content increased rapidly from 25 to 35 days after pollination（DAP）, and then decreased at 40 DAP. Triploid and tetraploid fruit had higher levels of lycopene than diploid. Moreover, triploid tended to contain more lycopene than tetraploid during fruit growth and ripening stages. However, little amount of lycopene（0–2 mg kg–1 fresh weight（FW）） in yellow-fleshed cultivar Huangmei was found during all fruit development stages. In Mimei, transcript level of PSY1 was generally higher than the other four genes, and LCYB gene expression was the lowest among all five genes being tested. PSY1, CRTISO and LCYB genes showed higher transcript levels in polyploid than in diploid fruit. By contrast, in Huangmei, transcript level of LCYB was not the lowest, but only lower than that of PSY1. PSY1, CRTISO and LCYB genes showed higher expression levels in diploid than in polyploid fruit. In Mimei, the negative correlation between gibberellane（GA） content and lycopene accumulation was determined in all three different ploidy fruits, while a positive correlation was observed between abscisic acid（ABA） content and lycopene accumulation only in diploid watermelon. These results indicated that different lycopene contents in different ploidy watermelons is regulated by the differential transcription expression of the lycopene metabolic genes and phytohormones.

Characterization of ploidy levels in Chrysanthemum L. by flow cytometry

Analyzing the ploidy levels of plants is important for identifying species, selecting parental lines, identifying the relationships between species, and determining evolutionary patterns. The genus Chrysanthemum is widely distributed throughout the world and exhibits different ploidy levels. We used flow cytometry to analyze the ploidy levels of nine species of Chrysanthemum L. collected from different regions and geographical locations in China. Three diploids from Henan and Wuhan provinces corresponded to Chrysanthe- mum lavandulifolium and two species of C. nankingense, while three tetraploids from various regions corresponded to C. indicum and two species of C. chanetii. Two hexaploids corresponding to C. vestitum were collected at Funiu moun- tain （Henan province）, and C. zawadskii was collected at Huangshan mountain （Anhui province）. We found that OTTO extraction buffer was suitable for extracting nuclei from most species, apart from C. zawadskii. Flow cytometry proved to bea simple, rapid, and highly accurate method for identifying ploidy levels in Chrysanthemum species.

Study on the Association Between Helicobacter Pylori Infection and the Pathogenesis of Gastric Cancer by Using Molecuar Biological Techniques

Recent epidemiological observations have indicated that the role of Helicobacter pylori (HP) infection in the pathogenesis of gastric cancer is an important but unresolved issue. We used the polymerase chain reaction, a sensitive and specific assay, to detect the HP infection in gastric biopsy specimens and examined the corrclations between HP infection and point mutation at 12 codon of H-ras oncogeneHras oncogene is higher in the group with HP infection than in those without HP infection. Furthermore, there is strong evidence that HP infection is associated with the increased expression of ras p21 protein , which suggested that HP infection increased the risk of ras oncogene activation. It is also noted that a significant relationship between infection with HP and the increase of DNA content and s% phase cells, indicated that rapid turnover of cells resulting from infection injury increases the risk of DNA damage.

Improved Detection of Cervical Cancer and High Grade Neoplastic Lesions by a Combination of Conventional Cytology and DNA Automated Image Cytometer

OBJECTIVE: To reduce false-negative rates of population based cervical screening programs employing conventional cytology in combination with automated DNA image cytometer. METHODS: Involved cervical samples from a total of 3603 women were taken by a cervix brush and then placed into a fixative solution. The cells were separated from mucus by mechanical and chemical treatment after which they were deposited onto microscope slides by a cytospin. Two slides were prepared from each case;one slide was stained by Papanicolaou stain for conventional cytology examination, while the other slide was stained by a DNA specific and stoichiometric stain. The latter slide was used to determine the relative amount of DNA in the cell nuclei in order to assess the ploidy status of the epithelial cells. Enrolled in the study, 157 women were followed by colposcopy examination where punch biopsies were taken from the visible lesions or from suspicious areas. The results of the conventional cytology were then compared to the DNA image cytometer for all samples. RESULTS: Histopathology diagnosed 51 lesions from the 132 biopsied cases as CIN2 or higher, including 27 CIN2, 16 CIN3 and 8 invasive cancers. Conventional cytology correctly identified 29 of the 51 high grade CIN and in-vasive cancer, while DNA image cytometer correctly identified 38 high grade CIN and invasive cancer using the crite-rion that at least three cells were found on the slide that contained DNA amount in excess of 5c. 42 out of 51 high grade CIN and invasive cancer were found by conventional cytology in combination with DNA image cytometer. Sensitivities were 56.8%, 74.5% and 82.4%, while specificities were 86.2%, 81.5% and 81.5% in conventional cytology, DNA image cytometer and combination both cytology and DNA image cytometer respectively. CONCLUSION: The study demon-strated that screening for high grade neoplastic lesions and cervical cancer by DNA image cytometer or combination of conventional cytology and DNA image cytometer is more sensitive than conventional screening approach.

THE CLINICAL PATHOLOGY AND DNA PLOIDY OF GASTRICMUCOSA ASSOCIATED LYMPHOID TISSUE (MALT)LYMPHOMA INFILTRATING THE LEIOMYOMASOF THE UTERUS

Objective: To investigate the clinicopathology and DNA ploidy of gastric mucosa associated lymphoid tissue (MALT) lymphoma infiltraving the leiomyomas of the uterus of a patient. Methods: The routine paraffin slides were cut, stained with HE, immunochemically by ABC methodusing the and stained by Feulgen method. Then the DNA ploidy of tumor cells was measured with an image cytometer. Results: In the mucosa, submucosa and the smooth muscle layer of the stomach and in the leiomyomas of the uterus there was diffusive and dense infiltration of centrocyte-like cells. The DNA measurement results were that the distribution of DNA mass of lymphoma cells in stomach and in lymph nodes had a single main aneuploidy peak each, and the distribution of DNA mass of lymphoma cells in leiomyomas of uterus had two peaks; one of them was the diploid, the other aneuploid. Conclusion: The MALT lymphoma cell invasion in uterus must be differentiated with a primary lymphoma in the uterus, the chronic lymphocyte leukemia in uterus and an endometrial stromal sarcoma. The present prognosis of the patient under discussion was poor. The follow-up results indicated the DNA index seemed to be important for predicting the malignancy degree and prognosis.

PROGNOSTIC SIGNIFICANCE OF DNA PLOIDY IN PATIENTS WITH STAGE II COLORECTAL CANCER

Objective: To evaluate the relationship between flow cytometric DNA ploidy, biological features and prognosis in patients with Stage II colorectal cancer. Methods: Nuclear DNA content, proliferation index and S-phase fraction were measured in a prospective series of 45 patients with curatively resected Stage II colorectal adenocarcinomas by means of flow cytometry using frozen tumor samples. Results: Of the 45 samples examined, 17 tumors (38%) were diploid and 28 (62%) aneuploid. The diploid tumors were significantly more common in the proximal colon than in the distal colon (67% vs. 23%; P<0.01). There was no correlation between DNA ploidy and the other clinicopathological variables (P>0.05). The proliferation index and S-phase fraction in the distal tumors were higher than those in the proximal tumors, but the difference was not significant (P>0.05). When the 5-year survival rate of patients with Stage II colorectal cancer was compared by the log rank test, a significant relationship between DNA ploidy status and disease free survival was observed in the group of all patients. Patients with DNA diploid tumors had a better disease free survival than those with DNA aneuploid tumors (P<0.05). Conclusion: These findings support that DNA ploidy status, proliferation index and S-phase fraction may differ in the proximal and distal colorectal cancer. Flow cytometric DNA ploidy status might be a useful prognostic factor in patients with Stage II colorectal cancer.