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1.
Chinese Journal of Forensic Medicine ; (6): 453-456,461, 2017.
Article in Chinese | WPRIM | ID: wpr-666645

ABSTRACT

Objective Construct a mRNA multiplex amplification system to identify different types of semen stains. Methods First, collect normal, oligozoospermia and azoospermia semen samples to make semen stains. Second, extract total RNA with Qiagen RNeasy Micro Kit. Then use reverse transcript PCR to amplify goal mRNA markers: 2 markers for sperm(PRM1, PRM2), 2 markers for seminal plasma(TGM4, SEMG1) and 2 housekeeping genes(TEF, UCE). Results All semen mRNA markers can be detected in normal semen samples. The RFU of sperm mRNA markers are lower in oligozoospermia semen samples than that in normal controls. No sperm mRNA markers can be detected in the azoospermia semen samples, only seminal plasma specific can be detected. Conclusion The differentiation of normal and azoospermia semen can be achieved by using multiplex mRNA fluorescence amplification system. While normal semen and oligozoospermia semen compared to no statistical difference.

2.
Article in Chinese | WPRIM | ID: wpr-503296

ABSTRACT

The identiifcation of body lfuid and their tissue resource is an important part of forensic medicine research. Conventional methods have imperfections like high false positive rate and may destroy biomaterial, which should be replaced by a new conifrmatory test. Highly-differentiated cells express speciifc mRNA molecules that can be used for body lfuid identiifcation. These mRNA markers can identify body lfuids like peripheral blood, menstrual blood, semen, saliva, vaginal secretion, skin-contact stains. This method have favorable sensitivity, speciifcity and can be detected after years, made it a ideal way to identify current body lfuids in the future. This review focus on the application of mRNA in body lfuid identiifcation and tissue resource.

3.
Article in Chinese | WPRIM | ID: wpr-391921

ABSTRACT

Objective To investigate the significance of genomic amplification of the telomerase RNA component (TERC) gene to serve as a genetic biomarker in the screening of cervicallesions.Methods A total of 715 cases were recruited,with liquid-based cytology diagnosis as normal (n=347),atypical squamous cells of undetermined significance (ASCUS,n=180),atypical squamous cells cannot exclude a high-grade lesion (ASC-H,n=13),low-grade squamous intraepithelial lesions (LSIL,n=115),high-grade squamous intraepithelial lesions(HSIL,n=59)and atypical glandular cells(AGC,n=1).The remaining cervical cells in the cytological preserving fluid were analyzed using a two-color fluorescence in situ hybridization (FISH) probe targeted to chromosome 3q26 containing TERC gene.The TERC gene findings were compared to the cytological and histological detected results,as well as high-risk human papillomavirus (HPV) detected results.Results Genomic amplification of TERC gene was found in 5.8% of normal specimens,22.2% of ASCUS.30.8% of ASC-H,27.8% of LSIL,86.4% of HSIL and 1/1 of AGC.The positive rate was significantly lower in normal,ASCUS,ASC-H and ISIL.compared with HSIL(all P<0.01).Significantly more cells with genomic amplification of TERC gene were found in cervical intraepithelial lesion(CIN) Ⅱ-Ⅲ than CIN Ⅰ (77.8% vs.9.3%),as well as invasive cervical cancer (96.7% vs.9.3%).both P < 0.01.The rate of TERC gene amplification was higher in HPV positive patients (33.5%) than in HPV negative patients(5.2%,P<0.01).The sensitivity of TERC gene amplification was significantly higher than that of cytological screening (81.88% vs.36.96%,P<0.01) in the differentiation of CIN Ⅱ or higher and CIN Ⅰ or lower diseases,its specificity Was hisher than high-risk HPV test (93.32% vs.33.93%,P<0.01) and positive prediction value (81.29%) was similar with cytological method (86.44%,P>0.05);but its negative prediction value (93.56%) was lower than HPV test (97.06%,P<0.05).Conclusions The positive rates of TERC gene amplification increased as cervical diseases worsened.TERC gene amplification is related to HPV infection.The gain of chromosome 3q26 in cytological specimens is an effective molecular genetic biomarker in screening of CIN Ⅱ or higher and invasive cervical cancer.

4.
Article in Chinese | WPRIM | ID: wpr-400364

ABSTRACT

ObjectiveTo study the value of liquid-based cytology and colposcopy in screening for cervical lesion among urban community women of Beijing.MethodsA total of 795 women aged 20~54 years with sexual activity living in Zhanlanlu Community of Beijing were screened for cervical lesion Cervcal specimen was collected for thin-layer,liquid-based cytology test (LCT) from each of the participants in gynecologic examination.Colposcopy and biopsy were performed for the women with positive LCT.ResultsForty-five of 795 (5.7%) women were positive for LCT[≥ASC-US (atypical squamouscell of undetermined significance)],with 33 of ASC-us,eight of low-grade squamous intraepithelial lesion (LSIL),three of high-grade squamous intraepithelial lesion (HSIL),one of atypical glandular cells (AGC).Five of 45 women with positive LCT refused to accept colposcopy.Among 40 women with colposcopy and biopsy,chronic cervicitis was diagnosed in 11(27.5%),cervical condyloma acurninatum in 14(35.0%),cervical intraepithelial neoplasia (CIN) 1 in seven(17.5%),CIN 2 in three (7.5%),CIN 3 in four(10.0%),and early invasive cervical cancer in one(2.5%).In 750 women with negative LCT,cervical condyloma acuminature was diagnosed in two(0.3%),CIN 1 in five(0.7%)and low-grade glandular intraepithelial lesion (LGIL) bin one(0.1%).Sensitivity and specificity of LCT screening for cervical lesion(≥CIN 1)were 71.4%and 94.2%,respectively,with positive and negative predictive values of 37.5%and 99.2%.respectively,and those screening for cervical lesion more than CIN 2 were 100.0%, 96.0%,20.5%and 100.0%,respectively.ConclusionsMore attention should be paid to early screening for cervical lesion in urban community women.LCT combined with colposcopy and biopsy provide very helpful information in screening for early cervical cancer.

5.
Article in Chinese | WPRIM | ID: wpr-397700

ABSTRACT

Objective To investigate the genomic amplification of the human telomerase RNA component (hTERC) gene in cervical cytology and evaluate its role in screening of cervical lesions. Methods A total of 301 cases were recruited, with liquid-based cytology diaghoses as normal (n=203), atypical squamous cells (ASC, n=66), low-grade squamous intraepithelial lesions ( LSIL,n=18), and high-grade squamous intraepithelial lesions ( HSIL, n=14). Following cytological examination, the slides were analyzed using a two-color fluorescence in aitu hybridization ( FISH ) probe targeted to chromosome 3q26 containing hTERC. The hTERC findings were compared to the cytologic and histologie results, as well as high-risk human papilloma viruses (HPV) results. Results Genomie amplification of hTERC was found in 3.0% (6/203)of normal specimens, 21.2% (14/66) of ASC, 44.4% (8/18) of LSIL and 92.9% (13/14) of HSIL, with a significant difference in each pair wise (all P<0.05). Significantly more cells with 3q26 gain were found in cervical intraepithelial lesion (CIN) Ⅱ than in CIN Ⅰ(75.0% vs. 20.0% ), as well as in CIN Ⅲ (86.7% vs. 20.0% ) and squamous cervical cancer (SCC) than in CIN Ⅰ (100.0% vs. 20.0%) ( all P<0.01). The sensitivity of hTERC amplification was significantly higher than cytological screening (82.6% vs. 17.4%, P<0.01), and its specificity was higher than high-risk HPV test (67.8%-73.5% vs. 25.6%-27.7%, P<0.01) in the diagnosis of HSIL (CIN Ⅱ - Ⅲ). The abnormal hTERC signal type mostly was 2:3 in CIN Ⅰ (84.9% ) ; whereas in CIN Ⅱ-Ⅲ, 2: 3, 2:4 and 4:4 accounted for 44.6%, 24.8% and 17.8%, respectively. Conclusion Testing the gain of chromosome 3q26 in cytological specimens using specific probe for hTERC is powerful in screening of HSIL, and the amplification patterns of 2:4 and 4:4 may serve as potential prognosis markers.

6.
Chinese Journal of Epidemiology ; (12): 947-950, 2007.
Article in Chinese | WPRIM | ID: wpr-322866

ABSTRACT

<p><b>OBJECTIVE</b>To observe the correlation between human papillomavirus (HPV) infection and cervical lesion among women living in community of Beijing.</p><p><b>METHODS</b>A total of 795 women at age 20-54, living in Zhanlanlu District of Beijing were screened for cervical lesion. Samples of cervical cytology (LCT) and HPV test (hc2) were collected. Colposcopy and biopsy were conducted in women with positive LCT.</p><p><b>RESULTS</b>In those 795 women, the infection rate of HPV was 14.1% (112/795). In 40 women who were LCT positive 1 early invasive cervical cancer, 4 cervical intra-epithelial neoplasia (CIN3), 3 CIN2 and 7 CIN1 were noticed. In 750 women with negative LCT, 5 CIN1 and 1 low-grade CGIN were diagnosed. In those women who were Cyto(+) and HPV(+), 15 cases (55.6%, 15/27) were diagnosed with > or = CIN1 (including 7 CIN1, 3 CIN2, 4 CIN3 and 1 early invasive cancer).</p><p><b>CONCLUSION</b>The risk of cervical lesion significantly increased in women showing positive in cytology and HPV test.</p>


Subject(s)
Adult , Biopsy , Cervical Intraepithelial Neoplasia , Diagnosis , Epidemiology , Virology , Cervix Uteri , Virology , China , Epidemiology , Colposcopy , Female , Humans , Middle Aged , Papillomavirus Infections , Epidemiology , Risk Factors , Uterine Cervical Diseases , Epidemiology , Uterine Cervical Neoplasms , Diagnosis , Epidemiology , Virology , Vaginal Smears , Young Adult
7.
Article in Chinese | WPRIM | ID: wpr-570348

ABSTRACT

Objective We study the regulation of human progestrone receptor isoforms A and B in uterine endometrial carcinoma cell line by different concentration of human insulin like growth factor Ⅰ (IGF Ⅰ) for different time,to investigate the roles of IGF Ⅰ and progestrone receptor isoforms in uterine endometrial carcimoma Methods The uterine endometrial adenocarcinoma cell line HEC IB was cultured in vitro and the breast cancer cell line MCF 7 was used as control Western blot was applied to examine the changes of the two isoforms by different concerntration IGF Ⅰ for different time Results (1) In HEC IB cell line, 10 ng/ml IGF Ⅰ made hPRB up regulated in the first 24 h But according to lager concerntration and longer time, human progesterone receptor (hPR) B became down regulated, which were significant at 20 ng/ml IGF Ⅰ for 72 h and 40 ng/ml IGF Ⅰ for 48-72 h The change of hPRA was like hPRB (2) In MCF 7 cell line, 10 ng/ml and 40 ng/ml IGF Ⅰ made hPRA and hPRB significantly up regulated in 24, 48, 72 h Twenty ng/ml IGF Ⅰ made hPRB up regulated also in the first 24 h But in 48 h and 72 h, down regulation of hPRB was detected Twenty ng/ml IGF Ⅰ made hPRA down regulated in 24, 48, 72 h Conclusions (1) The regulation of IGF Ⅰ to hPR isoforms has cell type specific and dose dependenct and time dependenct (2) In HEC IB cell line, 10 ng/ml IGF Ⅰ made hPRB significantly up regulated in 24 h But following exposure to IGF Ⅰ at larger concentration and longer time, hPRB became down regulated The change of hPRA is like hPRB

8.
Article in Chinese | WPRIM | ID: wpr-569675

ABSTRACT

Objective To investigate the role of two isoforms of human progesterone receptor A (hPR-A) and B(hPR-B) in the development of uterine leiomyoma, their distribution and expression in leiomyoma were detected. Methods The tissues of leiomyoma and normal myometrium from 30 uteri, which were excised for leiomyoma, were used for the localization and quantification of protein of the two isoforms. Immunohistochemistry and Western blot were applied respectively. Results Both hPR-A and hPR-B were nuclear receptors. Concentrations of hPR-(A+B) and hPR-A in leiomyoma were higher than those in normal myometrium (295 796? 90 856,256 275?98 560; P =0 042, P =0 000 563). Both isoforms were presented in leiomyoma and normal myometrium, with a consistent dominance of hPR-B over hPR-A. More expression of hPR-A was found during secretive phase than proliferative phase, not only in leiomyoma but also in normal myometrium ( P =0 037, P =0 024). Conclusions The development of uterine leiomyoma seems to be related with the progesterone receptor isoforms, especially hPR-A.

9.
Microbiology ; (12)1992.
Article in Chinese | WPRIM | ID: wpr-685745

ABSTRACT

Obtaining soil bacterial DNA of good quality is a key step in soil bacterial ecology study.A quick, efficient,sensitive and stably method of DNA extraction from soil were established by combining strongpoints of two kits(Soilmaster kit and DNA IQ~(TM)kit).In addition,the 16S rDNA gene and T-RFLP(Terminal restriction fragment length polymorphism)were used in the analysis of soil bacterial community diversity and the result show that T-RFLP is a powerful tool for bacterial community study.

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