[Analysis of a Chinese pedigree affected with Hereditary Fâ « deficiency due to compound heterozygous variants of F12 gene].

OBJECTIVE: To explore the laboratory phenotype and molecular pathogenesis in a Chinese pedigree affected with Hereditary coagulation factor Ⅻ (FⅫ) deficiency. METHODS: A male proband admitted to Ningbo No.2 Hospital on July 17, 2021 due to chronic gastritis and members of his pedigree (7 individuals from three generations) were selected as the study subjects. Prothrombin time (PT), activated partial thromboplastin time (APTT), FⅧ activity (FⅧ: C), FⅨ activity (FⅨ: C), FⅪ activity (FⅪ: C), FⅫ activity (FⅫ: C), and FⅫ antigen (FⅫ: Ag) were determined. All of the exons, exon-intronic boundaries, as well as the 5'- and 3'-untranslated regions of the F12 gene were subjected to Sanger sequencing. Candidate variants were verified by cloning sequencing. The effect of candidate variants on the protein function was analyzed by bioinformatics software. RESULTS: The proband, a 47-year-old male, had significantly prolonged APTT (180.0 s) and decreased FⅫ:C and FⅫ:Ag levels (< 1%). His father, mother, brother and two sons also showed certain degrees of reduction. Genetic testing revealed that the proband has harbored compound heterozygous variants of the F12 gene, namely c.1092_1093insC (p.Lys365Glnfs*69) in exon 10 and c.1792_1796delGTCTA (p.Val579Hisfs*32) in exon 14. His mother and elder son were heterozygous for the c.1092_1093ins variant, whilst his father, brother, and younger son were heterozygous for the c.1792_1796delGTCTA variant. Analysis of the promoter region of exon 1 also showed that the proband and both sons had harbored a 46T/T polymorphism, whilst other family members were 46C/T. Bioinformatic analysis suggested that the p.Val579 is a highly conserved site. Protein model analysis showed that, with the p.Val579Hisfs*32 variant, a benzene ring was added and the hydrogen bond of surrounding amino acids was changed. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1792_1796delGTCTA was classified as a pathogenic variant (PVS1+PM2_Supporting+PM4). CONCLUSION: The c.1092_1093insC (p.Lys365Glnfs*69) and c.1792_1796delGTCTA (p.Val579Hisfs*32) compound heterozygous variants of the F12 gene probably underlay the decreased FXII levels in this pedigree. Above finding has also enriched the mutational spectrum for FⅫ deficiency.

Roles of DANCR/microRNA-518a-3p/MDMA ceRNA network in the growth and malignant behaviors of colon cancer cells.

BACKGROUND: The competing endogenous RNA (ceRNA) networks of long non-coding RNAs (lncRNAs) and microRNAs (miRs) have aroused wide concerns. The study aims to investigate the roles of lncRNA DANCR-associated ceRNA network in the growth and behaviors of colon cancer (CC) cells. METHODS: Differentially expressed lncRNAs between CC and paracancerous tissues were analyzed using microarrays and RT-qPCR. Follow-up studies were conducted to evaluate the correlation between DANCR expression and prognosis of CC patients. Loss-of-functions of DANCR were performed to identify its role in the malignant behaviors of CC cells. Sub-cellular localization of DANCR and the potential targets of DANCR were predicted and validated. Cells with inhibited DANCR were implanted into nude mice to evaluate the tumor formation and metastasis in vivo. RESULTS: DANCR was highly-expressed in CC tissues and cell lines, and higher levels of DANCR were linked with worse prognosis and less survival time of CC patients. Silencing of DANCR inhibited proliferation, viability, metastasis and resistance to death of CC cells. DANCR was found to be sub-localized in cytoplasmic matrix and to mediate murine double minute 2 (MDM2) expression through sponging miR-518a-3p in CC cells, during which the Smad2/3 signaling was activated. Likewise, silencing of DANCR in CC cells inhibited tumor formation and metastasis in vivo. CONCLUSION: This study provided evidence that silencing of DANCR might inhibit the growth and metastasis of CC cells through the DANCR/miR-518a-3p/MDM2 ceRNA network and the defect of Smad2/3 while activation of the p53 signaling pathways. This study may offer novel insights in CC treatment.

Long Non-Coding RNA HCG11 Aggravates Osteosarcoma Carcinogenesis via Regulating the microRNA-579/MMP13 Axis.

BACKGROUND: Previous studies have suggested that long non-coding RNAs (lncRNAs) were involved in tumorigenesis of various human carcinomas, including osteosarcoma (OS). However, the expression and specific role of lncRNA HLA complex group 11 (HCG11) in OS remain unknown. The current study aimed at revealing the role of lncRNA HCG11 and its related mechanism in OS. METHODS: lncRNA HCG11 expression was verified with RT-qPCR followed by sub-localization determination. LncRNA-microRNA (miRNA) and miRNA-mRNA interactions were predicted by online bioinformatics websites. Validation was performed using dual-luciferase reporter gene assays, and gain- and loss-of-function experiments. The effects of lncRNA HCG11, miR-579 and matrix metalloproteinase 13 (MMP13) on the proliferation, migration and invasion, epithelial-mesenchymal transition (EMT) of OS cells were detected using cell counting kit-8 (CCK-8), Transwell assays and Western blot analysis. RESULTS: LncRNA HCG11 overexpression was observed in OS tissues and cell lines. Downregulation of lncRNA HCG11/MMP13 or overexpression of miR-579 blocked the progression of OS cells. LncRNA HCG11, which is located in the cytoplasm, promoted MMP13 expression through sponging miR-579. CONCLUSION: LncRNA HCG11 might be beneficial for OS aggravation via sponging miR-579 and facilitating MMP13 expression, which represents a candidate biomarker and target for OS therapy.

MicroRNA­466 inhibits cell proliferation and invasion in osteosarcoma by directly targeting insulin receptor substrate 1.

Accumulating evidence has demonstrated that microRNAs (miRNAs) are frequently dysregulated in osteosarcoma (OS), and the aberrant expression of miRNAs is associated with OS initiation and progression. Previous studies demonstrated that miRNA­466 (miR­466) is dysregulated, and serves important roles in various types of human cancer. However, the role of miR­466 in the formation and progression of OS remains unclear. In the present study, the expression level of miR­466 was identified to be markedly downregulated in OS tissues and cell lines. Additionally, miR­466 overexpression inhibited the proliferative and invasive abilities of OS cells. In the present study, bioinformatics analyses and luciferase assays were employed to show that miR­466 was able to directly target the 3'­untranslated region of insulin receptor substrate 1 (IRS1) gene, negatively regulating the mRNA and the protein expression levels of IRS1 in OS cells. Furthermore, IRS1 was upregulated in OS tissues, and the increased expression level of IRS1 exhibited an inverse correlation with the expression level of miR­466. Furthermore, IRS1 overexpression was able to partially reverse the suppressive effects of miR­466 overexpression in OS cells. To the best of the authors' knowledge, the present study is the first to suggest that miR­466 is downregulated in OS and inhibits the progression of OS by directly targeting IRS1. The present results suggested that miR­466 may represent a novel potential therapeutic target for the treatment of patients with OS.

[Effect and risk analysis of misoprostol in stimulating cervical maturity for post-term pregnancy].

OBJECTIVE: To evaluate the effect and risk of misoprostol for stimulating cervical maturity in women with post-term pregnancy negative for insulin-like growth factor binding protein-1 (IGFBP-1) in cervical secretion with modified Bishop score less than 3. METHODS: Seventy-one women with post-term pregnancy randomized into misoprostol group (n=37) and control group (n=34) received misoprostol placement at the posterior vaginal fornix and routine intravenous oxytocin infusion, respectively, to stimulate cervical maturity. Failure to respond to the treatment within the initial 24 h necessitated a repeated administration for no more than 3 times in all. Modified Bishop score was recorded and fetal heart monitored once every 24 h, and IGFBP-1 in the cervical secretion was detected at 24 and 48 h after drug administration. RESULTS: The misoprostol group showed better effect of cervical maturity stimulation than the control group (P<0.001), and the positivity rates of IGFBP-1 24 and 48 h after drug administration were significantly higher than that of the control group (P<0.01 and 0.001). The number of cases with indication for cesarean section was significant higher in the control group (P<0.001). There were no significant differences in postpartum hemorrhage, excessive uterine contraction, incidence of fecal contamination of the amniotic fluid or Apgar score of the newborn between the two groups (P>0.05). CONCLUSIONS: Misoprostol is safe and effective for stimulating cervical maturity in women with post-term pregnancy who have modified Bishop score lower than 3 and are negative for IGPBF-1 in cervical secretion. Oxytocin is not advised for use in such gravida for stimulating cervical maturity. IGFBP-1 in cervical secretion may serve as an important index for evaluating the cervical maturity.

[Characteristics of uterine contraction and stages of labor under continuous epidural block anesthesia].

OBJECTIVE: To observe the characteristics of uterine contraction and stages of labor during delivery under continuous epidural block anesthesia. METHODS: Totaling 213 parturients in spontaneous labor under epidural block anesthesia with dilated cervical orifice of 3 cm were monitored for the contraction cycle, duration, intensity and curve types of uterine contraction, and recordings were made for 30 min before and 30, 60 and 120 min after the anesthesia took effect, respectively. The duration of the active phase in the first, second and third stages of labor was compared between 421 cases with anesthesia and 237 without anesthesia. RESULTS: Significant difference was noted in the objective indexes of uterine contraction recorded after anesthesia had taken effect (P<0.05) in comparison with those before anesthesia, suggesting significantly attenuated uterine contraction after anesthesia, whereas these indexes underwent no significant further variation as compared between different time points after anesthesia (P>0.05). The average active phase in the first stage was significantly shorter in anesthesia group than that in the control group (P<0.05), but the average duration of the second and third stages of labor differed little between the two groups with appropriate use of oxytocin under strict monitoring (P>0.05). The rates of obstetric forceps utilization and use of oxytocin were higher in anesthesia group than in the control group (P<0.05). CONCLUSION: Epidural block anesthesia produces certain influences on uterine contraction and stages of labor during delivery, for which appropriate treatment measures may prove beneficial.