ABSTRACT
Objective To establish the blood Septin9 methylation detection system for early screening of colorectal cancer based on fluorescence PCR technology .Methods The PCR primer of Septin9 was designed by searching the CpG island site of Septin9 methylation in the NCBI database .The high methylation site of Septin9 gene promoter region was confirmed by PCR amplification and sequencing after extracting DNA from colorectal cancer and para-carcinoma tissues .The fluorescence PCR and TaqMan probe detection technique was designed by aiming at high methylation site for constructing the plasma sample methylation detection sys-tem .Then its accuracy ,specificity ,repeatability and minimum detectable amount were performed the assess-ment and analysis .The SETP9 methylation detection was performed in plasma samples from 57 cases of color-ectal cancer and 30 healthy persons .Results The high methylation site of Septin9 gene in tissue samples was confirmed by sequencing .This site served as the target for designing fluorescence PCR detection system .After assessment ,the accuracy ,specificity and repeatability of this detection system were 100％ ,the lowest detection amount reached 0 .1 ng/μL .Among the plasma samples in 57 cases of colorectal cancer ,the positive rate of Septin9 methylation site detection was 71 .92％ (41/57) and the positive rate of in the patients with pathologi-cal stage Ⅰ and Ⅱ of colorectal cancer reached 64 .2％ .But Septin9 gene had no methylation in plasma of healthy population .Conclusion The plasma fluorescence PCR detection system with Septin9 gene methylation as the target has the characteristics of high sensitivity ,high specificity and high accuracy ,which is the reliable detection technique for the early screening of colorectal cancer and has good clinical application prospect .
ABSTRACT
Objective To investigate the significance of the combined detection of several serum tumor markers with high-risk human papillomavirus (HR-HPV) in the diagnosis of cervical cancer.Methods Chemiluminescent microparticle immunoassay(CMIA) was employed to measure the levels of serum CA125,CA19-9,SCCA and CEA in peripheral blood samples collected from 249 patients with cervical cancer,30 patients with cervix intraepithelial neoplasia(CIN),and 60 healthy controls.The levels of serum CA72-4,HE4 were measured by electrochemiluminescence immunoassay(ECLI).Flow-through hybridization and gene chip technology (HybriMax) was employed to measure the HPV genotypes in all the 339 cases of women.Results Serum CA72-4 level in the cervical cancer group [3.13 (2.25,7.63) ng/mL] was significantly higher than that in the C IN group [2.37 (1.98,6.25) ng/mL] and the control group [2.69 (2.35,3.35) ng/mL] (P =0.028;P =0.003).Serum CEA level in the cervical cancer group 3.08 (2.28,4.75) ng/mL was significantly higher than that in the CIN group [1.45 (1.00,1.83) ng/mL] and the control group [2.13 (1.45,2.67) ng/mL] (P =0.000;P =0.000).Serum CA 125 level in the cervical cancer group [24.4(20.30,44.15) U/mL] was significantly higher than that in the CIN group[12.85(7.90,16.23)U/mL] and the control group[12.33 (11.26,17.11) U/mL] (P =0.000;P =0.000).Serum CA19-9 level in the cervical cancer group[27.05(18.48,38.01) U/mL] was significantly higher than that in the CIN group[13.01 (8.79,17.12) U/mL] and the control group[12.83 (10.89,17.93) U/mL] (P =0.000;P =0.000).Serum SCCA level in the cervical cancer group [1.7 (0.90,4.75) ng/mL] was significantly higher than that in CIN group [0.8 (0.60,1.10) ng/mL] and the control group [0.6 (0.50,0.70) ng/mL] (P =0.000;P =0.000).However,there was no significant difference of HE-4 among three groups (P ＞ 0.05).The HR-HPV infection rates in the cervical cancer (92.4％) and CIN patients (90.0％) were significantly higher than that in controls (15.0％)(x2 =46.875,P =0.000;x2 =165.178,P =0.000).In the cervical cancer patients,the levels of serum tumor marker was associated with Federation International of Gynecology and Obstetrics (FIGO) stage (P ＜ 0.05).ROC curves showed HR-HPV was the most sensitive in the diagnosis of cervical cancer.The sensitivity of the diagnosis of cervical cancer by serum tumor markers combined with HR-HPV(97.99％) was significantly higher than that of single marker,CA724 25.30％,CEA 20.89％,CA125 27.71％,CA199 24.90％,SCCA 52.61％,HPV 92.37％ (x2 =278.237,P =0.000;x2 =307.036,P =0.000;x2 =263.348,P =0.000;x2 =280.769,P =0.000;x2 =137.864,P =0.000;x2 =8.580,P =0.003).Conclusion Serum tumor markers combined with HR-HPV check can improve the detection rate of cervical cancer and has important clinical value.
ABSTRACT
Objective To investigate the changes of CD4+ CD25+ CD127low regulatory T cells (Treg) in peripheral blood samples collected from the patients with cervical cancer and its clinical signifi -cance , and to evaluate the correlations between Treg cells and the infection of high-risk human papillomavir-us ( HR-HPV) .Methods Flow cytometry analysis was performed to measure the percentages of CD4+ CD25+ CD127low Treg cells among CD4+T cells in peripheral blood samples collected from 249 patients with cervical cancer , 30 patients with cervix intraepithelial neoplasia ( CIN) and 60 healthy subjects .The corre-lations between the levels of Treg cells and the clinicopathological features of cervical cancer were analyzed . Flow-through hybridization and gene chip technology ( HybriMax) were used to analyze the genotypes of HPV strains isolated from the 339 subjects.The correlations between Treg cells and HR-HPV infection were ana-lyzed.Results Compared with the healthy subjects , the patients with cervical cancer or CIN showed higher percentages of CD4+ CD25+ CD127low Treg cells and rates of HR-HPV infection (P<0.01).The percentages of CD4+ CD25+ CD127low Treg cells in patients with cervical cancer were significantly correlated with the sta-ges of cervical cancer , which was staged by the Federation International of Gynecology and Obstetrics (FIGO) staging system,and the degrees of differentiation (P<0.05).The percentages of CD4+ CD25+ CD127low Treg cells in the peripheral blood of patients with positive HR-HPV were significantly higher than those in patients without HR-HPV infection (P<0.01).Conclusion The CD4+ CD25+ CD127low Treg cells in peripheral blood might play an important role in the oncogenesis and development of cervical carcinoma , thus it could be used as a potential marker for the evaluation of disease progression .Moreover , the CD4+ CD25+ CD127low Treg cells were closely related to the HR-HPV infection.
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Objective To investigate the changes of CD 4+CD25+CD127 low regulatory T ( Treg ) cells in peripheral blood samples from patients with lung cancer and their correlation with clinicopathologic features of lung cancer .Methods Flow cytometry was used to measure the percentages of CD 4, CD8, nat-ural killer ( NK) and Treg cells in peripheral blood samples collected from 160 patients with lung cancer and 60 healthy subjects .The correlations between the levels of Treg cells and clinicopathologic features of lung cancer were analyzed .The percentages of Treg cells in 60 patients with lung cancer before and after surgery were compared .Results The percentages of CD 4+and NK cells and the ratios of CD 4+/CD8+cells in pa-tients with lung cancer were significantly lower than those in healthy control , while the percentages of Treg cells in patients with lung cancer were decreased as compared with those in healthy subjects .The percenta-ges of Treg cells in patients with advanced cancer were significantly higher than those in patients at early stage (P<0.05), which dropped significantly after surgery (P<0.01).Conclusion The results of this study indicated that Treg cells in patients with lung cancer might inhibit antitumor immune responses and correlate with the progression of cancer .It would be worthwhile to check Treg cells in patients with lung cancer for monitoring the prognosis and treatment effects .
ABSTRACT
Objective To explore a quick and feasible method of screening malaria parasite by using a Sysmex XE-2100 hematology analyzer though alarm information on high eosinophil count and atypical eosinophil distributions in the WBC scattergram. Methods Sysmex XE-2100 hematology analyzer was used for complete blood cell analysis. Microscopic review was used when high eosinophil count and atypicaleosinophil distributions in the WBC scattergram were found. If the review showed normal eosinophil cells, wecontinued to focus on red cell for searching malaria parasite. Results Among 1 501 specimens showing higheosinophil counts and atypical eosinophil distributions, nine cases with normal eosinophil cells were indisaccordance with the hematology analyzer, six of them showed high eosinophil count in the Sysmex XE-2100 hematology analyzer, whose distribution was located close to neutrophil clusters in scattergram. The otherthree had an abnormal WBC scattergram. There was no gap between eosinophil clusters and neutrophilclusters, which brought no classified results. But all the nine specimens had been found the trophozoite,schizont and merozoite in blood smears. Conclusions There were great possibility of the existence of themalaria parasite in specimens when hematology analysis showed high eosinophil count and atypical eosinophildistributions in the WBC scattergram in a Sysmex XE-2100 hematology analyzer, although these alarm wasnot comfirmed by microscopic review. This method provides not only a simple and reliable way for quickscreening malaria parasite but also has a great value in preventing the undetected ratio on malaria parasite.