Paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples / 浙江大学学报·医学版

OBJECTIVE@#To investigate paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples.@*METHODS@#p16 immunohistochemical staining was performed in 150 cases of chronic cervicitis, 126 cases of low-grade squamous intraepithelial lesions(LSIL), 96 cases of high-grade squamous intraepithelial lesions (HSIL) and 78 cases of cervical cancer from January 2014 to March 2018 in Zhongshan Boai Hospital. The results of p16 protein expression in paraffin sections with thickness of 2, 3, 4, 5 and 6 μm were compared using Logistic regression analysis.@*RESULTS@#With the increase of slice thickness, the staining of p16 protein in nucleus gradually increased. The thickness of cervical slices in chronic cervicitis and cervical cancer samples had no significant effect on the positive rate of p16 protein(=7.817 and 1.332, both >0.05), while the thickness of slices in LSIL and HSIL samples had significant effect on the positive rate of p16 protein (=17.688 and 10.182, <0.05 or <0.01). The stable and reliable results were obtained when the slices were between 3 and 5 μm thick.@*CONCLUSIONS@#Paraffin sections with thickness of 3.0-5.0 μm are recommended for immnohistochemical staining of p16 protein in cervical tissue samples.

Volume of 4% neutral buffered formaldehyde affects the results ofgene detected by fluorescencehybridization in primary invasive breast cancer / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of the volume of 4% neutral phosphate buffered formalin fixative solution on the detection of human epidermal growth factor receptor 2 () gene by fluorescencehybridization (FISH) in primary invasive breast cancer.</p><p><b>METHODS</b>Tissue samples were collected from 109 patients with primary invasive breast cancer admitted in Zhongshan Boai Hospital from June 2014 to October 2016. The ratios of 4% phosphate buffered formalin fixative solution to sample volume samples were 3:1, 6:1, 9:1, 10:1, 15:1, 20:1 or 25:1 (groups A, B, C, D, E, F and G), respectively. Paraffin sections were made after 15 h of fixation. The amplification ofgene was detected by FISH. The gene amplification results ofwere observed and compared in different groups.</p><p><b>RESULTS</b>Fluorescence microscope showed that the tissue contour in groups A, B and C was vague, cell debris appeared, and the probe was positioned poorly; while the tissue contour was clear and complete in groups D, E, F and G and the probe was positioned accurately. The positive rate ofwas gradually increased from group A to D(=8.601,<0.01), and that remained stable at 24.77% in groups D to G. The positive rate of gene amplification in groups D, E, F and G was significantly higher than that in groups A, B and C (all<0.05).</p><p><b>CONCLUSIONS</b>When using FISH to detectgene in samples of primary breast invasive carcinoma, the volume of fixative solution should be at least 10 times of the sample volume to obtain accurate and stable results.</p>

Application of polyhydroxy acrylic acid and Van-clear in the detection of EGFRgene mutations in non small-cell lung cancer by qRT-PCR method / 西安交通大学学报(医学版)

Objective To compare two different methods to detect the differences of gene mutation rate, sensitivity, specificity and coincidence rate of epidermal growth factor receptor (EGFR) in non-small-cell lung cancer (NSCLC) so as to assess the clinical value of qRT-PCR method and its environmental-friendly technologyplatforms.One uses environmental fixative poly hydroxyl acrylic acid and green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ (volume fraction) neutral buffered formalin and the traditional transparent dewaxing liquid xylene in application of quantitative real-time polymerase chain reaction (qRT-PCR).The other uses traditional reagents in direct sequencing.Methods We selected 91 cases of primary NSCLC specimens resected between May 2013 and March 2016 in Zhongshan Bo`ai Hospital and Zhongshan Hospital of Traditional Chinese Medicine.Five samples were taken from the same tumor lesion.We used a random number table to randomly divide these samples into Groups A, B , C, D, and E.Group A received direct sequencing method in detection of EGFR gene mutations.Besides, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group B received qRT-PCR method to detect EGFR gene mutations.Meanwhile, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group C received qRT-PCR method in detection of EGFR gene mutations.At the same time, during the experiment, polyhydroxy acrylic acid was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group D received qRT-PCR method to detect EGFR gene mutations.In the meantime, 4％ neutral buffered formalin was used for fixing, Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.Group E received qRT-PCR method in detection of EGFR gene mutations.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.The mutations of Exons 18, 19, 20, and 21 in EGFR genes were respectively determined in the five groups of NSCLC.Results ① Groups B, C, D, E and A did not significantly differ in the percentage of people with mutations or target site mutation rates of EGFR genes in NSCLC (P> 0.05).② The detection results of EGFR target site mutation in Groups B, C, D, E and A had good sensitivity, strong specificity, and high compliance rate.Conclusion The green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ neutral buffered formalin and the traditional transparent dewaxing liquid xylene in the application of qRT-PCR so as to detect EGFR gene mutations in NSCLC has good consistent results compared with the method that uses traditional reagents in direct sequencing.It has the significance and value in clinical application.

Application of Van-Clear and xylene in the detection of cervical hTERC gene by fluorescence in situ hybridization / 中南大学学报(医学版)

OBJECTIVE@#To observe the effect of Van-Clear on vamplification of human telomerase RNA component (hTERC) gene in cervical tissues by fluorescence in situ hybridization, and to determine the potential for Van-Clear to replace xylene.
@*METHODS@#A total of 278 specimens of cervix uteri were collected from inpatients of Department of Gynaecology in Boai Hospital of Zhongshan from January to February, 2015, with 81 cases of normal specimens, 68 cases of cervical intraepithelial neoplasia (CIN) I, 57cases of CIN2, 42 cases of CIN3 and 30 cases of cervical invasive cancer. Double samples were collected from the same region. Fluorescence in situ hybridization was applied to detect the changes in the amplification of hTERC gene in 2 groups of specimens from the cervical biopsy.
@*RESULTS@#Differences in the positive expression rate of hTERC gene between the 2 groups of cervical lesions at all levels were not statistically significant (P>0.05).
@*CONCLUSION@#There is no significant difference in the positive rate of hTERC gene expression between the slices made by Van-clear and xylene. As an environmental-friend product, Van-Clear possesses certain value in detection of cervical hTERC gene by fluorescence in situ hybridization.

Expression and significance of HSPC238 in cervical intraepithelial neoplasia and cervical cancer / 中国免疫学杂志

Objective: To investigate the expression of HSPC238 in cervical intraepithelial neoplasia and cervical cancer.Methods:We collected 76 cases of cervical cancer,105 cases of CIN and 28 cases of normal cervical epithelial.Then we inves-tigated the expression of HSPC238 by using immunohistochemistry and compared the significant differences between them.Results:There was no significant difference in the expression of HSPC238 between the cervical cancer and normal cervical epithelial ( Z=-0.242,P>0.05).However,there was significant difference between the cervical intraepithelial neoplasia and normal cervical epithelial (χ2=19.159,P<0.01) and the expression of HSPC238 was correlated with the grades of CIN.The expression of HSPC238 decreased when the grade of CIN was increasing.( rs=-0.327,P<0.01 ).Conclusion:The low expression of HSPC238 might be correlated with the development of cervical neoplasia.

Comparison of the role that poly hydroxy acrylic acid plays in the detection ofHER-2 gene in breast cancer by hematoxylin and eosin staining and fluorescencein situ hybridization with that of 4% neutral buffered formaldehyde / 中国癌症杂志

Background and purpose: Adequate tissue ifxation, transparent dewaxing is an important step of hematoxylin eosin (HE) staining and lfuorescence in situ hybridization (FISH) in detection of breast cancerHER-2 gene. The purpose of this study was to make a comparison between poly hydroxyl acrylic acid which is an environmen-tally friendly ifxation liquid and 4% neutral buffered formaldehyde in tissue ifxation for HE staining and FISH to detect theHER-2 gene in the breast cancer tissue sections. The study aimed to evaluate the feasibility of replacing 4% buffered formaldehyde, a traditional ifxation liquid, with the poly hydroxyl acrylic acid, an environmentally friendly ifxation lfuid.Methods:This project was performed on tissue samples collected from 69 cases of breast cancer, 41 cases of breast ifbroadenoma, 40 cases of uterine leiomyoma, 25 cases of cervical tissue, 25 cases of placenta obtained from the outpatient and inpatient departments of Zhongshan Boai Hospital from Mar. 2011 to Jan. 2015, from each of which two samples were drawn and two blocks of each specimen were divided into two groups randomly. Then one group was ifxed with 4% neutral buffered formaldehyde and made into 200 sections by HE while the other group was ifxed with poly hydroxyl acrylic acid and made into another 200 sections. The slice level of the two groups was determined by the staining condition of the sections, and SPSS 19.0 was employed to compare the excellent and good rate of HE staining. Additional 69 sections were produced with two groups of breast invasive ductal carcinoma tissues, and SPSS 19.0 was used to detect the ampliifcation ofHER-2 gene by FISH.Results:First, the number of best-quality slices stained with HE ifxed separately by poly hydroxyl acrylic acid and 4% neutral buffered formaldehyde was 155 and 166, respectively. The number of excellent pieces was 41 and 33, respectively, while the number of mediocre pieces was 3 and 1 with bad pieces being 1 and 0, respectively. The excellent and good rates of HE staining were 98% and 99.5%, respectively. There was no significant difference between the two groups (χ2=1.33,P>0.05).Second, the positive rates of the tis-sue slices by FISH ifxed separately by poly hydroxyl acrylic acid and 4% neutral buffered were 26.09% and 23.19%, respectively. There was no signiifcant difference between the two groups (χ2=0.50,P>0.05).Conclusion:The results obtained with HE staining and FISH using poly hydroxyl acrylic acid as a ifxation liquid are not signiifcantly different from those using 10% neutral buffered formaldehyde. Therefore, poly hydroxyl acrylic acid meets the requirements of environmental protection, and thus has the potential to be promoted and widely used.

Comparison between poly hydroxy acrylic acid and Van-clear replacing the tradi-tional reagents to detect the cervical hTERC genes by adopting FISH technique / 北京大学学报(医学版)

Objective:To observe the difference of the human telomeres RNA component (hTERC) genes’amplification in the cervical tissue by applying the environment-friendly fixative poly hydroxy acrylic acid and the transparent dewaxing solution Van-clear separately or jointly to replace the traditional fixative 4% (volume fraction)neutral buffered formalin and the conventional transparent dewaxing solu-tion xylene in the use of fluorescence in situ hybridization (FISH)for detection.Methods:In the study, 255 cases of cervical tissue specimens submitted by the Department of Gynecology in Zhongshan Boai Ho-sipital were collected from Mar.2013 to Apr.2015.Four samples were taken from the same lesion site. All the cases were divided into 4 groups and named group A,B,C,and D.Group A used 4% neutral buffered formalin fixed and xylene dewaxing to make slices.Group B used poly hydroxy acrylic fixed and xylene dewaxing to make slices.Group C used 4% neutral buffered formalin fixed and Van-clear trans-parent to make slices.Group D used poly hydroxy acrylic fixed and Van-clear transparent dewaxing to make slices.The amplification of hTERC genes in the four groups of cervical specimens was also detected by FISH technique.Results:When the hTERC genes were detected by FISH method under the fluore-scence microscope,it was obvious that the tissue profile and the background of group A,B,C and D were all clear.The probe was fixed in the accurate position so that the bright red or green fluorescence signals were easily found in these four groups.Compared with the positive rate of group A,there was no statistical significance in that of group B,C and D (P>0.05).At the same time,the coincidence rate of the FISH results was high,which showed that the new environment-friendly reagent had no significant difference in the detection of cervical hTERC genes by FISH technique.Conclusion:It is possible for the environment-friendly reagent poly hydroxy acrylic acid and Van-clear to replace 4%neutral buffered for-malin and xylene separately or jointly to detect the cervical hTERC genes by adopting FISH technique.

Clinical analysis of cervical screening in 2329 pregnant women / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the cytopathologic characteristics of cervical diseases in pregnant women and the outcomes of the postpartum women to provide evidence for prevention and treatment of cervical cancer.</p><p><b>METHODS</b>This study was conducted among 2329 pregnant women undergoing routine gestational examinations between September, 2012 and September, 2013. The women with abnormal cytological findings by Thin-prep cytology test (TCT) were followed up and colposcopy and cervical biopsy were performed. The TCT results of these women were compared with those of 32 491 non-pregnant women in Zhongshan Cervical Cancer Mass Screening Program.</p><p><b>RESULTS</b>Of the 2329 pregnant women, a total of 97 patients had abnormal TCT results (4.16%). Cervical biopsy were performed for 14 patients (14.43%), and 8 (57.14%) of them had evidence of cervical intraepithelial neoplasia (CIN) or cancer on biopsy. In the 32491 non-pregnant women in the mass screening program, 1383 (4.26%) women had abnormal TCT results and cervical biopsy were performed for 248 patients (17.93%), among whom 148 (59.68%) had evidence of CIN or cancer on biopsy. The rate of high-grade squamous intraepithelial lesion (HSIL) was significantly higher in non-pregnant women than in pregnant women (P=0.033), but the total rate of cytological abnormalities were comparable between them (P=0.911). The patients with CIN had regular examinations during pregnancy and postpartum follow-up showed no invasive carcinoma.</p><p><b>CONCLUSION</b>Pregnancy is not a risk factor to accelerate the progress of cervical lesions, and most of the cervical lesions are relieved or show no progression in the postpartum women, suggesting the feasibility of follow-up during pregnancy and postpartum reevaluation for patients with CIN in pregnancy.</p>

Application Study of Thinprep Liquid Based Cytology Test in the Detection of Cervical Diseases / 中国医师杂志

Objective To assess the value of thinprep liquid based cytology test (TCT) and Besthesda system (TBS) in diagnosis of cervial lesions. Methods 11000 cases of cervical lesions were detected with TCT and TBS.The samples of atypical squamous cells(ASC) and more severe than ASC lesions were classified into postive cytology,and then cytological results were compared with pathological examination results of cervical biopsies.Results The satisfactory rate of TCT samples was 96.36%, and the postive rate of abnormal epithelial cells diagnosed by TCT was 10.96%(1206/11000). Among 1206 cases, 15 cases(0.14%) of CA, 89 cases (0.81%) of HSIL, 449 cases (4.08%) of LSIL, 73 cases (0.66%) of ASC-H and 580 cases (5.27%) of ASC-US and AGUS. 1128 cases with postive cytologic results received pathological examination. The pathological positive rate of CA, HSIL, LSIL, ASC-H, and ASC-US and AGUS were 100%, 100%, 67.12%, 54.17% and 14.09%, respectively. The coincident rate of two examination methods was 100% for CA, 91.1% for HSIL,57.82% for LSIL and 27.78% for ASC-H. Conclusion TCT and TBS can accurately detect cervical cancer, precancerous lesions and infection of bacteria, virus and other pathogens, and at present is a means of screening cervical cancer.