1.Analysis of karyotype and copy number variation in 386 fetuses with increased nuchal translucency
Yan TANG ; Shoulian LU ; Shengnan SONG ; Jue WANG ; Mingzhu MIAO
Chinese Journal of Clinical Laboratory Science 2025;43(2):98-101
Objective To investigate the karyotypes and the correlation of chromosomal abnormalities in the fetuses with increased nu-chal translucency(NT),so as to provide a basis for prenatal genetic counseling.Methods The clinical data of 386 singleton pregnant women with NT≥2.5mm who underwent invasive prenatal diagnosis at the First Affiliated Hospital of Nanjing Medical University from January 2018 to April 2022 were retrospectively analyzed.The fetuses were grouped according to NT thickness(2.5-3.4,3.5-3.9,4.0-4.9,5.0-5.9,and ≥6.0 mm),fetal ultrasound abnormalities(isolated increased NT,non-isolated increased NT),and maternal age(advanced age ≥35 years,non-advanced age<35 years).The chi-square test was used to compare the differences of the incidence of fetal chromosomal abnormalities among various groups.Results Among the 386 fetuses with increased NT,chromosomal abnormalities were detected in 87 cases with an overall detection rate of 22.5%(87/386),including chromosomal numerical abnormalities accounted for 82.8%(72/87)and copy number variations(CNVs)accounted for 17.2%(15/87).The detection rates of chromosomal abnormal-ities and numerical abnormalities increased with NT thickness(P<0.05),while no statistically significant difference of CNV abnormali-ty rates was found(P=0.41).The detection rates of chromosomal abnormalities(36.5%)and CNV abnormalities(14.1%)in the non-isolated increased NT group were significantly higher than those in the isolated increased NT group(18.6%and 1.0%,respective-ly,both P<0.05).The detection rates of chromosomal abnormalities(34.7%)and numerical abnormalities(31.6%)in the fetuses of advanced maternal age mothers with increased NT were significantly higher than those in the non-advanced age group(18.4%and 14.2%,respectively,both P<0.05).However,the difference of CNV abnormality rates between the two groups was not statistically sig-nificant(P=0.62).Conclusion The detection rate of fetal chromosomal abnormalities elevated with increased NT thickness.Ad-vanced maternal age and the presence of other ultrasound abnormalities were the high-risk factors for fetal chromosomal abnormalities.The risks of CNV abnormalities may not be significantly correlated with NT thickness or maternal age but associated with the presence of other ultrasound abnormalities.
2.Analysis of karyotype and copy number variation in 386 fetuses with increased nuchal translucency
Yan TANG ; Shoulian LU ; Shengnan SONG ; Jue WANG ; Mingzhu MIAO
Chinese Journal of Clinical Laboratory Science 2025;43(2):98-101
Objective To investigate the karyotypes and the correlation of chromosomal abnormalities in the fetuses with increased nu-chal translucency(NT),so as to provide a basis for prenatal genetic counseling.Methods The clinical data of 386 singleton pregnant women with NT≥2.5mm who underwent invasive prenatal diagnosis at the First Affiliated Hospital of Nanjing Medical University from January 2018 to April 2022 were retrospectively analyzed.The fetuses were grouped according to NT thickness(2.5-3.4,3.5-3.9,4.0-4.9,5.0-5.9,and ≥6.0 mm),fetal ultrasound abnormalities(isolated increased NT,non-isolated increased NT),and maternal age(advanced age ≥35 years,non-advanced age<35 years).The chi-square test was used to compare the differences of the incidence of fetal chromosomal abnormalities among various groups.Results Among the 386 fetuses with increased NT,chromosomal abnormalities were detected in 87 cases with an overall detection rate of 22.5%(87/386),including chromosomal numerical abnormalities accounted for 82.8%(72/87)and copy number variations(CNVs)accounted for 17.2%(15/87).The detection rates of chromosomal abnormal-ities and numerical abnormalities increased with NT thickness(P<0.05),while no statistically significant difference of CNV abnormali-ty rates was found(P=0.41).The detection rates of chromosomal abnormalities(36.5%)and CNV abnormalities(14.1%)in the non-isolated increased NT group were significantly higher than those in the isolated increased NT group(18.6%and 1.0%,respective-ly,both P<0.05).The detection rates of chromosomal abnormalities(34.7%)and numerical abnormalities(31.6%)in the fetuses of advanced maternal age mothers with increased NT were significantly higher than those in the non-advanced age group(18.4%and 14.2%,respectively,both P<0.05).However,the difference of CNV abnormality rates between the two groups was not statistically sig-nificant(P=0.62).Conclusion The detection rate of fetal chromosomal abnormalities elevated with increased NT thickness.Ad-vanced maternal age and the presence of other ultrasound abnormalities were the high-risk factors for fetal chromosomal abnormalities.The risks of CNV abnormalities may not be significantly correlated with NT thickness or maternal age but associated with the presence of other ultrasound abnormalities.
3.Influence of CD133+expression on patients' survival and resistance of CD133+cells to anti-tumor reagents in gastric cancer
Dehu CHEN ; Ruiqi LU ; Xiaochun NI ; Jugang WU ; Shoulian WANG ; Bojian JIANG ; Jiwei YU
Asian Pacific Journal of Tropical Biomedicine 2015;(12):996-1004
Objective: To investigate the influence of CD133+expression on patients' survival and resistance of CD133+cells to anti-tumor agents in gastric cancer (GC).
Methods: Influence of CD133 expression on prognosis was analyzed employing sam-ples from patients with GC. GC cell lines were utilized to separate CD133+and CD133?subpopulations by immunomagnetic separation and to analyze the biological features of two subpopulations in vitro and in vivo, especially in resistant to anti-tumor reagents and its apoptotic mechanism.
Results: The lower CD133+group showed a significantly better survival compared with the higher CD133+group. The highest content of CD133+subpopulations for KATO-III cells had stronger proliferative ability than CD133?subpopulations. A single CD133+cell was capable of generating new cell colony and the tumorigenicity rate in nude mice was 100% for CD133+ clonal spheres or for CD133+ cells, but 0% for CD133? cells. Furthermore, the higher expression levels of Oct-4, Sox-2, Musashi-1 and ABCG2 in CD133+ clonal spheres were identified compared with CD133+ cells or CD133? cells. Under the treatment of anti-tumor reagents, CD133+ cells had lower suppression rates compared with CD133? cells while lower level of Bcl-2 and higher level of Bax were found in CD133+cells compared with CD133?cells.
Conclusions: The patients with lower CD133+expression had a better survival. Enriched CD133+ cells in clonal sphere shared the ability to be self-renewable, proliferative, tumorigenic and resistant to anti-tumor agents as probably regulated by Bcl-2 and Bax.
4.Progress of stem cell surface marker CD133 in gastric cancer
Shoulian WANG ; Ruiqi LU ; Bojian JIANG
International Journal of Surgery 2012;39(1):56-59
The morbidity and mortality of gastric cancer stand in the first place among all kinds of malignant tumours in our country,the early diagnosis rate is still very low and the prognosis is unsatisfactory also.The expression of CD133 probably as a tumour stem cell marker is significantly higher in gastric cancer primary lesion than that in normal tissue and its higher expression corelates with the larger size of tumor,lymph node metastasis occurences and poorer prognosis.Meanwhile,the positive expression of CD133 may have some relationship with the angiogenesis,severer infiltration depth,worse differentiation degree and later TNM stage of gastric cancer.This paper aims at further recongnizing the specific expression of CD133 in gastric cancer primary lesion,both the relationship with initiating and clinical pathological features of gastric cancer,so as to bring a new research direction for early diagnosis and targeted therapy of stomach cancer.
5.Interference of RNAi to CD133 gene and the comparison of the interferential effects in KATO-Ⅲ cells of human gastric cancer
Shoulian WANG ; Jiwei YU ; Ruiqi LU ; Cheng CAI ; Jugang WU ; Xiaochun NI ; Bojian JIANG
International Journal of Surgery 2012;39(11):755-759,封4
Objective To compare the inhibition effects of three synthesized fragments used in small interfering RNA(siRNA) against CD133 gene in KATO-Ⅲ gastric cancer cells,and to study effects of suppressed CD133 on the proliferating ability of intervened cells.Methods Three fragments of siRNA were designed and synthesized targeted at the mRNA of CD133.Cell fluorescence counting under confocal laser scanning microscope was used to determine the transfection efficiency after transfection with the CD133FITC-siRNA.The knock-down effect of the CD133 gene was detected by RT-PCR and Western blotting.CCK-8 (cell counting kit-8 assay) was performed to measure the variation of the cell proliferative viability after the above-mentioned treatment.Results The transfection efficiency of siRNA was (85 ± 8) % in KATO Ⅲ Gastric cacer cell.All these three fragments of CD133 siRNA effectively inhibited the expression of CD133 gene,the inhibition rate being (11 ± 2) %,(19 ± 2) %,(24 ± 3) %respectively.Compared with the control group,the cell proliferation viability was restrained (42 ± 4)% in CD133siRNA-3 group (P <0.05).Conclusions CD133siRNAs were successfully transfected into KATO Ⅲ Gastric cacer cells and repressed the expression of CD133.Meanwhile,the CD133siRNA fragment 3 was screened from three CD133 siRNA,which has the best inhibition effect.The results provide preliminary evidence for the intereference of CD133+ gastric cancer cells subsequently.
6.Transforming growth factor-β1 generates epithelial-to-mesenchymal transition and promote CD44 expression in SGC7901 cells
Cheng CAI ; Jiwei YU ; Jugang WU ; Ruiqi LU ; Xiaochun NI ; Shoulian WANG ; Bojian JIANG
International Journal of Surgery 2012;39(11):-
Objective To investigate the biological effect of epithelial-to-mesenchymal transition (EMT) under the treatment of transforming growth factor (TGF)-β1 on the human gastric cancer cell line SGC7901 in vitro,and to observe whether the TGF-β1 can generate the tumor initiating cells ability in SGC7901 or not.Methods SGC7901 cells were cultured with TGF-β1.The morphological change was observed.The effect on proliferation of SGC7901 cells was detected by CCK-8.The invasion assay was used to investigate the motility and the invasion ability of SGC7901 cells.Immuofluorescence was used to detect the expression of E-cadherin and N-cadherin.The mRNA and protein's expression levels of EMT-related factors and CD44 were analyzed by RT-PCR and Western blotting respectively.Results TGF-β1 induced morphological alterations from epithelial to mesenchymal cells.The proliferation of SGC7901 cells was inhibited,and the ability of motility and invasion of SGC7901 cells were greatly enhanced after being treated with TGF-β1.RT-PCR and Western blotting showed that the expression of Snail (P < 0.05),N-cadherin (P < 0.05) and CD44 (P < 0.05) were significantly increased while the expression of E-cadherin was decreased (P < 0.05).Conclusions TGF-β1 can generate the EMT.The CD44 expression was up-regulated.TGF-β1 can inhibit the proliferation and promote the motility and invasion ability of SGC7901 cells.
7.Transforming growth factor-β1 induces epithelial-to-mesenchymal transition and promotes abtaining of stemness characteristics in gastric cancer
Cheng CAI ; Jiwei YU ; Jugang WU ; Ruiqi LU ; Xiaochun NI ; Shoulian WANG ; Bojian JIANG
International Journal of Surgery 2012;(12):824-829,封3
Objective To investigate if TGF-β1 induces epithelial-mesenchymal transition (EMT) and promotes the obtaining of stemness characteristics in gastric cancer cell lines.Methods After KATO-Ⅲ cells were cultured with or without 5 ng/mL TGF-β1,the morphological change was observed and compared under phase-contrast microscopy.At the same time,the effect of TGF-β1 on the proliferation of KATO-Ⅲ cells was detected by CCK-8.On the other hand,the mRNA and protein' s expressions of EMT-related factors,ESC markers and TICs markers were analyzed by RT-PCR and Western blotting methods too.Results TGF-β1 induced morphological alterations from epithelial to mesenchymal cells.The proliferation of KATO-Ⅲ cells was inhibited after treated with TGF-β1 (P < 0.05).After treated with TGF-β1,the relative mRNA expression levels of Snail (0.5219 ±0.0147) and N-cadherin(0.6640 ±0.0124) were higher than that in control group(0.2049 ±0.0214,P =0.004,0.2722 ± 0.0098,P =0.001),the relative protein expression levels of Snail (0.4769 ± 0.0234) and N-cadherin (0.5014 ± 0.0216) were higher than that in control group (0.2534 ± 0.0345,P =0.02,0.2026 ± 0.0268,P =0.009),while the relative E-cadherin mRNA and protein levels in TGF-β1 treated group (0.4701 ± 0.0215,0.1349 ± 0.0258) were lower than that in control group (0.6792 ± 0.0157,P =0.01 ; 0.6055 ± 0.0227,P =0.004),while the relative mRNA expressions of ESC markers such as Sox2,OCT4,Nanog in TGF-β1 treated group (0.594 ± 0.039、0.438 ± 0.033、0.489 ± 0.037) were higher than that in control group (0.143 ± 0.013,P =0.001,0.156 ± 0.025,P =0.001,0.325 ± 0.046,P =0.03),the relative mRNA expression levels of CD44 (0.437 ±0.037) and CD133(0.543 ±0.028) were higher than that in control group (0.247 ±0.024,P =0.000,0.139 ± 0.016,P =0.000),the relative protein expression levels of CD44 (0.429 ± 0.034) and CD133 (0.316 ±0.027) in TGF-β1 treated group were higher than that in control group (0.152 ± 0.014,P =0.000,0.110 ±0.010,P =0.000),cloning sphere-forming capacity was greatly enhanced after treated with TGF-β1 (P < 0.01).Conclusion TGF-β1 can induce EMT in KATO-Ⅲ cells and promote the obtaining of stemness characteristics in gastric cancer cell lines.

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