Objective:To study the effects of Hedgehog signal transduction pathway on the cell proliferation, apoptosis and connexin 32 (Cx32)and connexin 43 (Cx43)membranous distribution of breast cancer cells,and to explore its mechanism in the cell proliferation and metastasis of breast cancer.Methods:The breast cancer MCF-7 cells at logarithmic growth period were divided into cyclopamine groups and blank control groups. The MCF-7 in cyclopamine groups were treated with 5,10,20,30 and 40μmol·L-1 for 24,48 and 72 h;MTT assay was applied to detect the inhibitory rate of proliferation of MCF-7 cells. After the MCF-7 cells were treated with 0 (negative control group)and 25μmol·L-1 cyclopamine for 48 h,flow cytometry was employed to determine the apoptotic rate and to analyze membranous distribution of Cx32 and Cx43 in the MCF-7 cells.Results:Compared with blank control group,the inhibitory rates of proliferation in cyclopamine groups were increased (P<0.05), and the inhibitory effect of proliferation was increased with the increasing of cyclopamine doses and prolongation of treatment time.After treated with 25μmol·L-1 cyclopamine,the apoptotic rate of MCF-7 cells was higher than that in blank control group (P<0.05).The positive expression rates of Cx32 and Cx43 48 h after treatment were higher than those in negative control group (P<0.05).Conclusion:Hedgehog signal transduction pathway can inhibit the apoptosis and mediate membranous distribution of Cx32 and Cx43 in breast cancer cells.

Objective To investigate the gene transduction efficiency of lentiviral vector in leukemia cells to provide key basis for leukemia gene therapy. Methods A third-generation self-inactivating(SIN) lentiviral vector system based on human immunodeficiency virus type 1(HIV-1) was used to improve transduction efficiency.The transduction efficiency of the HIV-1-based vector was compared directly with the moloney murine leukemia virus(MLV) SIN vector in human leukemia cell line K562.The expression of green fluorescent protein(GFP) in cells was observed by fluorescence microscopy and flow cytometry(FCM) to detect the percentage of gene trasduction cells.Results The GFP expression in K562 cells was observed qualitatively by fluorescence microscopy.At the same gene transduction conditions,the GFP marker gene expression intensity and GFP positive cells in leukemia cells transduced with HIV vectors were significantly higher than those transduced with MLV vectors.Initial transduction efficiencies were almost 100% for the HIV and less than 40% for the MLV vectors. The transduction efficiency had significant difference between HIV vector group and MLV group(P

Objective To investigate chromosome abnormality rate and related factors of spontaneous abortion in early pregnancy. Methods A total of 831 tissue samples of spontaneous abortion in early pregnancy were collected from June 2015 to August 2018 in the First Affiliated Hospital of Nanjing Medical University. Chromosomal copy number was analyzed by next generation sequencing (NGS). The relationships between chromosome abnormality and maternal age, in vitro fertilization?embryo transfer (IVF?ET) pregnancy, number of previous spontaneous abortions, history of live birth were analyzed by statistical methods. Results Among 831 tissue samples of spontaneous abortion in early pregnancy, 461 (55.5%, 461/831) were found to have chromosome abnormalities. Maternal age (OR=1.107, 95%CI: 1.070-1.145) and history of live birth ( OR=1.909, 95%CI : 1.182-3.083) were the positive correlative factors of chromosome abnormality. Times of previous spontaneous abortion (OR=0.807, 95%CI: 0.702-0.928) and IVF?ET pregnancy ( OR=0.554, 95%CI : 0.404-0.760) were the negative correlative factors of chromosome abnormality. Conclusions Chromosome abnormality is an important cause of spontaneous abortion in early pregnancy. The rate of chromosome abnormality increases with the increase of maternal age and the history of live birth, and decreases with the increase of number of previous spontaneous abortion and IVF?ET pregnancy.

Objective: To investigate chromosome abnormality rate and related factors of spontaneous abortion in early pregnancy. Methods: A total of 831 tissue samples of spontaneous abortion in early pregnancy were collected from June 2015 to August 2018 in the First Affiliated Hospital of Nanjing Medical University. Chromosomal copy number was analyzed by next generation sequencing (NGS). The relationships between chromosome abnormality and maternal age, in vitro fertilization-embryo transfer (IVF-ET) pregnancy, number of previous spontaneous abortions, history of live birth were analyzed by statistical methods. Results: Among 831 tissue samples of spontaneous abortion in early pregnancy, 461 (55.5%, 461/831) were found to have chromosome abnormalities. Maternal age (OR=1.107, 95%CI: 1.070- 1.145) and history of live birth (OR=1.909, 95%CI: 1.182-3.083) were the positive correlative factors of chromosome abnormality. Times of previous spontaneous abortion (OR=0.807, 95%CI: 0.702-0.928) and IVF-ET pregnancy (OR=0.554, 95%CI: 0.404-0.760) were the negative correlative factors of chromosome abnormality. Conclusions Chromosome abnormality is an important cause of spontaneous abortion in early pregnancy. The rate of chromosome abnormality increases with the increase of maternal age and the history of live birth, and decreases with the increase of number of previous spontaneous abortion and IVF-ET pregnancy.

OBJECTIVE: To explore the genetic etiology of recurrent hydatidiform mole (RHM) and provide accurate guidance for reproduction. METHODS: Peripheral venous blood samples of the probands with RHM and members from 5 unrelated pedigrees were collected. Genomic DNA was extracted by using routine method, and whole exome sequencing was carried out to detect variants of RHM-associated genes including NLRP7 and KHDC3L. Sanger sequencing and real-time quantitative PCR (RT-qPCR) were used to validate the candidate variants and delineate their parental origin. RESULTS: Homozygous or compound heterozygous variants of the NLRP7 gene were identified in four patients from three pedigrees, which included a homozygous deletion of exon 1 to 4 of NLRP7 in patient P1 and her elder sister, compound heterozygous variants of NLRP7 c.939delG (p.Q314Sfs*6) pat and c.1533delG (p.N512Tfs*4) mat in patient P2, and compound heterozygous variants of NLRP7 c.2389_2390delTC (p.A798Qfs*6) pat and c.2165A>G (p.D722G) mat in patient P4. All variants were interpreted as pathogenic or likely pathogenic according to the American College of Medical and Genomics (ACMG) guidelines. Among these, NLRP7 exons 1 to 4 deletion, c.939delG (p.Q314Sfs*6), c.1533delG (p.N512Tfs*4) and c.2389_2390delTC (p.A798Qfs*6) were unreported previously. CONCLUSION Variants of the NLRP7 gene probably underlay autosomal recessive RHM in the three pedigrees, and definitive molecular diagnosis is beneficial for accurate genetic counseling. Above finding has also enriched the spectrum of the NLRP7 variants underlying RHM.
Adaptor Proteins, Signal Transducing/genetics* ; Aged ; China ; Female ; Homozygote ; Humans ; Hydatidiform Mole/pathology* ; Mutation ; Pedigree ; Pregnancy ; Sequence Deletion