Overlapping Cervical Cell Image Segmentation Based on Bottleneck Detection and Watershed Algorithm / 中国医疗器械杂志

This study proposes an image segmentation method based on bottleneck detection and watershed algorithm to solve the problem of overlapping cervical cell image. First, we use polygon approximation to get all feature points on the cell contour and then use bottleneck detection and ellipse fitting to obtain the correct split point pairs. Therefore, the approximate range of the overlapping region was determined. The watershed algorithm was used to obtain the internal boundary information for the gradient image of the region. Finally, the segmentation results of the overlapped cells were obtained by superimposing with the outer contour. The experimental results show that this algorithm can segment the contour of a single cell from the overlapping cervical cell images with good accuracy and integrity. The segmentation result is close to that of doctors' manual marking, and the segmentation result is better than other existing algorithms.

Effect of HPV detection and cervical cell basis detection on early diagnosis of cervical precancerous lesions / 中国基层医药

Objective@#To analyze the effect of human papillomavirus (HPV) detection and cervical cell basal fluid on the early diagnosis of cervical precancerous lesions.@*Methods@#From January 2016 to January 2017, 223 patients in the Maternal and Child Health Care Hospital of Yongkang who received gynecological cervical cancer precancerous lesions screening were selected.All patients into the outpatient screening categories were given cervical liquid-based cytology (TCT) examination and HPV detection, 61 patients with test results of double positive received colposcopy cervical biopsy histopathological examination, the final diagnosis of 49 patients with epithelial cell changes.The accuracy and sensitivity of different detection methods were compared.@*Results@#The detection rates of high risk HPV infection in cervicitis, CINⅠ, CINⅡ-CINⅢ and cervical cancer patients were 16.22%, 39.29%, 71.43% and 100.00% in 223 cases of this study.The total positive rate of TCT was 28.57%.The total positive rate was 44.44% in HPV group and 75.51% in combination test.The positive rate of combined detection was higher than TCT (χ2=21.63, P=0.000) and HPV (χ2=25.51, P=0.000), the difference was statistically significant.For ASC patients, the sensitivity of TCT, HPV and combined detection was 57.14%, 64.29% and 85.71%, respectively.The accuracy of TCT, HPV and combined detection was 64.29%, 71.43% and 92.86%, respectively.The sensitivity of combined detection was similar to TCT (χ2=2.13, P=0.140) and HPV (χ2=1.25, P=0.260). The accuracy of combined detection was similar to TCT (χ2=2.33, P=0.130) and HPV (χ2=1.39, P=0.240). For LISIL patients, the sensitivity of TCT, HPV and combined detection was 56.25%, 62.50% and 87.50%, respectively, the accuracy was 68.75%, 75.00% and 93.75%, respectively.The sensitivity of the combined test group was higher than that of TCT (χ2=3.86, P=0.049) and HPV (χ2=4.57, P=0.033), and the accuracy of combined test was higher than that of TCT (χ2=3.902, P=0.048) and that of HPV (χ2=4.13, P=0.034). For HISIL typing patients, the sensitivity of TCT, HPV and combined detection was 57.89%, 63.16% and 89.43%, respectively, and the accuracy was 73.68%, 78.95% and 94.74%, respectively.The sensitivity of the combined test group was higher than that of TCT (χ2=4.89, P=0.027) and that of HPV (χ2=3.99, P=0.047), and the accuracy of combined test was higher than that of TCT (χ2=3.99, P=0.048) and that of HPV (χ2=5.34, P=0.027).@*Conclusion@#High-risk HPV detection combined with TCT in the screen of cervical cancer precancerous lesions is simple and efficient, taking into account the accuracy and sensitivity, it can reduce the rate of missed diagnosis, has a positive effect on prevention and control of cervical cancer.

Effects of miR-1246 on proliferation,invasion and migration of cervical squamous cell carcinoma cell line SiHa and its target gene / 西安交通大学学报(医学版)

ABSTRACT:Objective To explore the effects of miRNA-1246 (miR-1246)on cell proliferation,invasion and migration in human cervical squamous cell carcinoma (CSCC)cell line SiHa.Methods SiHa cells were assigned into 3 groups:miR-1246 analog group,miR-1246 antagonist group and control group.Transfection efficiency was determined.The MTT assay,transwell assay and wound healing assay were performed respectively to evaluate the proliferation,invasion and migration abilities of SiHa cells.Western blot was carried out to detect the expression of thrombospondin-2 (THBS2)before and after transfection.A THBS2 3’-UTR-containing dual luciferase plasmid was synthesized and co-transfected with miR-1246 into SiHa cells to observe the luciferase enzyme activity.Results MTT assay,transwell assay and wound healing assay revealed that the abilities of proliferation,migration and invasion were significantly enhanced (P<0.01)in SiHa cells transfected with miR-1246 analog,but suppressed in SiHa cells transfected with miR-1246 antagonist.Western blot showed that SiHa cells transfected with miR-1246 analog had significantly decreased THBS2 expression (gray value = 6 .2 8 ± 1 0 .2 2 , P=0 .0 1 3 ) while those transfected with miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90±19.81, P=0.037).After co-transfected with miR-1246 and THBS2 3’-UTR-containing plasmid,SiHa cells exhibited a decreased level of luciferase enzyme expression.Conclusion miR-1246 promoted the proliferation,invasion and migration of CSCC SiHa cell, and it might promote CSCC tumorigenesis and progression by suppressing the expression of its target gene THBS2 .

Genotypes analysis of HPV infection in cervical cell samples among women in Xxuzhou region / 国际检验医学杂志

Objective To investigate the distribution situation of human papillomavirus (HPV) genotypes profile in cervical cells among women in Xuzhou area and its clinical significance .Methods 23 kinds of HPV DNA were extracted in cervical cell samples from 8 010 women in Xiuzhou area .The gene‐chips technique of PCR combined with reverse dot blot was adopted to detect the HPV genotypes .Results Among 8010 cervical cell samples ,there were 1 852 HPV infected cases ,the total HPV infection rate was 23 .12% ,the HPV infection rates of single type accounted for 17 .17% and its predominant types were 16 type (4 .35% ) ,followed by 58 type (2 .12% ) and 52 type (1 .82% ) ,The detection rate of multiple HPV infection was 5 .96% ,in which the predominant types were HPV16+58(4 .40% ) ,16+52(2 .94% ) ,11+16(2 .52% ) .Conclusion The single HPV infection of HPV16 ,58 ,52 and the multiple HPV infection of HPV16+58 ,16+52 ,11+16 are the main genotypes of cervical cells among women in Xuzhou area , this gene chip technique is suitable for the cervical cell sample ,its once detection can detect 23 kinds of HPV genotypes with high specificity and high sensitivity ,which has an important significance for the molecular epidemiologic survey study of HPV genotypes distribution among women in our country .

The anticancer effect and mechanism of photodynamic therapy using a new photosensitizer and diode laser on cervical cancer cell / 대한산부인과학회지

OBJECTIVE: During the last several years, such as Photofrin(R), Photogem(R), HpD, and 5-aminolevulenic acid (ALA) has been used as photosensitizers for a wide range of malignant tumors as well as non-malignant disease. A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a) was derived from Spirulina platensis. The aim of this study was to evaluate the anticancer effect and mechanism of photodynamic therapy in vitro using 9-HpbD-a and 670 nm diode laser on a HT-3 cervical cancer cell line. METHODS: We studied the cytotoxic effect of 9-HpbD-a and 670 nm diode laser in HT-3 cervical cancer cell line. The cultured HT-3 cells were treated with serial concentrations of 9-HpbD-a followed by various irradiation time (0, 5, 15, 30 min) and by various interval times (0, 3, 6, 9, 12 24 hours) until laser irradiation, then 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) assay was applied to measure the relative inhibitory effect of PDT. RESULTS: Optimal condition for laser irradiation time was 15 minutes and the cytotoxic effect according to the incubation time after 9-HpbD-a treatment was increased until 6 hours. Under confocal microscopy, to observe intracellular localization of 9-HpbD-a, cells dual-stained with 9-HpbD-a and panel of organelle-specific fluorescence probes (MitoTracker, LysoTracker, ER-Tracker), endoplasmic reticulum (ER) as a major accumulation organelle site in HT-3 cervical cell line for this new photosensitizer. CONCLUSION: The anticancer effect of PDT using 9-HpbD-a is effective in HT-3 cervical cancer cell line. This agent therefore represents a promising photosensitizing prodrug for the treatment of cervical cancer with PDT in clinical setting for the future.

In vivo study of photodynamic therapy using a new photosensitizer, 9-hydroxypheophorbide-a and 670 nm diode laser on HT-3 cervical cancer cell / 대한산부인과학회지

OBJECTIVE: During the last several years, such as Photofrin(R), Photogem(R), HpD, and 5-aminolevulenic acid (ALA) has been used as photosensitizers for a wide range of malignant tumors as well as non-malignant disease. A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a) was derived from Spirulina platensis. The aim of this study was to evaluate the anticancer effect and mechanism of photodynamic therapy in vivo using 9-HpbD-a and 670 nm diode laser on a HT-3 cervical cancer cell line. METHODS: PDT effects were observed in xenograft nude mouse model. HT-3 cell line was xenografted into the subcutaneous space of the mouse back. When the tumors grew up to 400~500 mm3, the animals were randomly separated into 4 groups: Group I (n=5), control group; group II (n=10), 9-HpbD-a alone; group III (n=10), laser irradiation alone; and group IV (n=20), PDT in nude mouse group. RESULTS: PDT treated group (9-HpbD-a+laser irradiation) showed complete response (15/20, 75%), recurrence (3/20, 15%) or no response (2/20, 10%). PDT group showed significant remission rate, comparing to laser irradiation alone group, 9-HpbD-a alone group and untreated control group (P<0.05). When PDT using 9-HpbD-a, tumor size was decrease and microscopically finding, antitumor effect was showed inflammation and necrosis of tumor cell. Overexpression of p53 and Ki-67 gene had been detected at the HT-3 cervical cancer cell and 10 days after PDT. And no experssion in normal tissue and tumor after PDT. So, immunohistochemical study of p53 and Ki-67 gene correlated with PDT for anticancer effect. CONCLUSION: These results suggest that 9-HpbD-a is one of the promising photosensitizers for the future and would be needed to use 9-HpbD-a as a photosensitizer in clinical setting. We would further suggest that photodynamic therapy is a more effective method than the conventional treatment for women who want to maintain their anatomical and functional uterine cervix.