BACKGROUND: Studies have shown that chitooligosaccharides have antitumor effect. However, the influence of chitooligosaccharides on cyclin D1, bcl-2 and bcl-xl remains unclear.OBJECTIVE: To observe the JnhJbJtJon effect of chitooligosaccharides on the proliferation of Hela cells, and the influence on cyclin D1, bcl-xl and bcl-2 mRNA expression.METHODS: Hela cells were stimulated by different concentrations of chitooligosaccharides (0.1, 0.5, 1, 2, 5 g/L). The effects of chitooligosaccharides on Hela cells were detected by CCK8 kit. Using real-time PCR methods, the gene expression of cyclin D1, bcl-xl and bcl-2 mRNA were determined.RESULTS AND CONCLUSION: Chitooligosaccharides inhibited the proliferation of Hela cells. With the concentrations of chitoolJgosaccharides increased from 0.1 g/L to 2 g/L, the inhibition effects on the gene expressJon of cyclin D1, bcl-xl and bcl-2 were enhanced, peaked at 2 g/L, and decreased at high concentration (5 g/L). Antitumor activity of chitooligosaccharides may exert through two aspects: it inhibits cyclin D1 mRNA expression to suppress the proliferation of tumor cells; on the other hand, chitooligosaccharides inhibits the expression of bcl-xl and bci-2 to promote the apoptosis of tumor cells. Moreover, the effects of the former are stronger than the latter.

OBJECTIVETo detect potential mutations of COL1A1 and COL1A2 genes with polymerase chain reaction-high-resolution melting analysis(PCR-HRMA) in a proband diagnosed with osteogenesis imperfecta (OI).

METHODSPeripheral blood samples were collected from the proband and members of his family as well as healthy controls. The mutations were detected by PCR-HRMA and confirmed by direct sequencing. Potential effects of the mutations were predicted using softwares including PolyPhen, SIFT and Align GVGD.

RESULTSThe PCR-HRMA has indicated mutations in exon 45 of the COL1A1 gene in the proband as well as his parents, which were presented as the difference in the melting curves between the patients and the control samples. Sequencing analysis confirmed that the proband has carried two heterozygous mutations (c.3235G>A, p.Gly1079Ser and c.3247G>A, p.Ala1083Thr) in exon 45 of the COL1A1 gene. Among them, c.3235G>A was predicted to have impeded alpha helix structure domain, which was inherited from the father who also had OI. c.3247G>A was inherited from mother who had a normal phenotype. All three softwares predicted that the c.3235G>A mutation can interfere with the function of the protein, while the c.3247G>A may have a benign effect by PolyPhen analysis.

CONCLUSIONThe study identified two mutations (c.3235G>A and c.3247G>A) occurred simultaneously in COL1A1 gene in a case. The case is the first reported in human collagen mutation database. As identified,mutation of c.3235G>A may be the major cause of the disease in the proband.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Child ; Child, Preschool ; China ; Collagen Type I ; genetics ; Exons ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Osteogenesis Imperfecta ; genetics ; Pedigree ; Point Mutation

Objective:To carry out genetic analysis on two families with carriers of small terminal translocations using karyotyping analysis and genomic copy number variation sequencing (CNV-seq).Methods:Two couples undergoing prenatal diagnosis at the Tianjin Central Hospital of Obstetrics and Gynecology respectively on April 12, 2020 and December 17, 2021 were selected as the study subjects. With informed consent, amniotic fluid and peripheral blood samples were collected and subjected to conventional karyotyping and CNV-seq analysis for the detection of chromosomal microdeletion/duplications.Results:Both couples had given births to children with chromosomal aberrations previously, and both fetuses were found to have abnormal karyotypes. CNV-seq showed that they had harbored microdeletion/duplications, and their mothers had both carried balanced translocations involving terminal fragments of chromosomes.Conclusion:For fetuses with small chromosomal segmental abnormalities, their parental origin should be traced, and the diagnosis should be confirmed with combined genetic techniques.

OBJECTIVETo explore genetic mutations and clinical features of osteogenesis imperfecta type V.

METHODSClinical record of five patients (including one familial case) with osteogenesis imperfecta type V were retrospectively analyzed. Peripheral blood samples of the patients, one family member, as well as healthy controls were collected. Mutation of IFITM5 gene was identified by PCR amplification and Sanger sequencing.

RESULTSA heterozygous mutation (c.-14C>T) in the 5-UTR of the IFITM5 gene was identified in all of the patients and one mother. The clinical findings included frequent fractures and spine and/or extremities deformities, absence of dentinogenesis imperfecta, absence of hearing impairment, and blue sclera in 1 case. Radiographic findings revealed calcification of the interosseous membrane between the radius-ulna in all cases. Hyperplastic callus formation was found in 3 cases. Four had radial-head dislocation.

CONCLUSIONA single heterozygous mutation c.-14C>T was found in the 5-UTR of the IFITM5 gene in 5 patients with osteogensis imperfecta type V. The patients showed specific radiological features including calcification of interosseous membrane, hyperplastic callus formation, and radial-head dislocation.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Male ; Mutation ; Osteogenesis Imperfecta ; diagnostic imaging ; genetics ; Young Adult

OBJECTIVE: To determine the combined cytotoxic effect and the molecular basis of triptolide and sodium cantharidinate on hepatoma cell line 7721.
 METHODS: After treating the hepatoma cell line 7721 with triptolide(9, 18, or 36 μg/mL) and/or sodium cantharidinate (2, 5, or 10 μg/mL), cell viability assay and apoptosis were examined by MTT and flocytometry, respectively. The protein levels of caspase 3 and nuclear factor κB were analyzed by Western blot.
 RESULTS: Viability of hepatoma cell line 7721 was inhibited by either the therapy of triptolide and/or sodium cantharidinate (P<0.05) in a time- and dose-dependent manner. The combined effects of both drugs were better than those of the single drug (P<0.05). The combined therapy down-regulated the expression of NF-κB p65 (P<0.05) while up-regulated the expression of caspase-3 (P<0.05).
 CONCLUSION Triptolide and sodium cantharidinate exert a synergistic toxic effect on hepatoma cell line 7721, which is related to increasing capase-3 activity and suppression of NF- κB.
Apoptosis ; drug effects ; Cantharidin ; pharmacology ; therapeutic use ; Carcinoma, Hepatocellular ; drug therapy ; Caspase 3 ; drug effects ; Cell Line, Tumor ; Diterpenes ; pharmacology ; therapeutic use ; Down-Regulation ; Drug Therapy, Combination ; Epoxy Compounds ; pharmacology ; therapeutic use ; Humans ; Liver Neoplasms ; drug therapy ; NF-kappa B ; drug effects ; Phenanthrenes ; pharmacology ; therapeutic use ; Transcription Factor RelA