Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.

OBJECTIVE: To carry out prenatal diagnosis for a fetus with increased nuchal translucency (NT) and another fetus with non-invasive prenatal testing (NIPT) suggested reduced sex chromosomes by cytogenetic and molecular techniques. METHODS: Chromosomal karyotyping, single nucleotide polymorphism array (SNP-array) and fluorescence in situ hybridization (FISH) were applied for the diagnoses. Peripheral blood samples were also taken from their parents for chromosomal karyotyping and SNP-array analysis. RESULTS: Both fetuses showed a 46,X,+mar/45,X karyotype. SNP-array has detected a 22.0 Mb duplication at Yp11.31q11.223 and a 3.9 Mb microdeletion at Yq11.223q11.23 in fetus 1, and a 16.9 Mb duplication at Yp11.31q11.221 and a 8.1 Mb deletion at Yq11.222q11.23 in fetus 2. The results were confirmed by FISH. The parents of both fetuses were normal by chromosomal karyotyping and SNP-array. CONCLUSION Combined use of various techniques can enable accurate prenatal diagnosis and genetic counseling.

OBJECTIVE: To carry out prenatal diagnosis for a fetus with absent nasal bone by using cytogenetic and molecular techniques. METHODS: Chromosomal karyotyping, single nucleotide polymorphism array (SNP-array) and fluorescence in situ hybridization (FISH) assays were applied for the diagnoses. Peripheral blood samples were also taken from the parents for chromosomal karyotyping and FISH analysis. RESULTS: The fetus was found to have a 46,XX,add(21)(p11.2) karyotype, and SNP-array has revealed a 11.3 Mb duplication at 21q22.12q22.3 (hg19: 36 762 648-48 093 361), which was confirmed by FISH. Both parents were found to be normal by chromosomal karyotyping and FISH analysis. The fetus was ultimately found to have a karyotype of 46,XX,der(21)t(21;21)(p11.2;q22.1), resulting a de novo partial trisomy of 21q22.1. CONCLUSION Combined use of various techniques has enabled accurate prenatal diagnosis and genetic counseling for the fetus.
Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Nasal Bone ; Pregnancy ; Prenatal Diagnosis ; Trisomy/genetics*

Objective To investigate the roles of ultrasound,laboratory methods,and genetic diagnostic techniques in screening and diagnosing fetuses with an unbalanced recombination of chromosome 18[rec(18)] due to parental pericentric inversion,and the relationship between rec(18) fetal phenotypes and their recombinant chromosomes.Methods We analyzed two pedigrees with pericentric inversion of chromosome 18 (including the fetuses and their parents) which received prenatal diagnosis and genetic counseling on March 2017 and March 2018 respectively at Xiamen Maternity and Children Health Care Hospital through karyotype analysis,chromosome microarray analysis(CMA) and fluorescence in situ hybridization (FISH).Literatures were retrieved from Scientific Citation Index,PubMed,China National Knowledge Infrastructure(CNKI) and Wanfang Data from 1970 to June 2018.The genetic counseling records,ultrasound and laboratory findings,pregnancy outcomes of families with pericentric inversion of chromosome 18 in this study and the included literatures were reported and analyzed.Results Non-invasive prenatal testing (NIPT) of one case indicated high risk of fetal trisomy 18 at 22 weeks of gestation.And the imaging examination indicated that fetus had interventricular septal defect and micrognathia at 24+2 weeks.Prenatal diagnosis confirmed that the fetal karyotype was 46,XY,rec(1 8)dup(18q) inv(18)(p1 1.32q12.1) pat,which was originated from his father whose karyotype was 46,XY,inv(1 8)(p1 1.32q12.1).In the other case,serum screening testing indicated high risk of fetal trisomy 18 at 12+3 weeks.Imaging examination indicated that fetus had thickened nuchal translucency at 13+3 weeks and bilateral choroid plexus cysts at 15+6 weeks.Prenatal diagnosis confirmed that the fetal karyotype was 46,XY,rec(18)dup(18q)inv(18) (p11.32q12.1) mat,which was originated from his mother whose karyotype was 46,XX,inv(18)(p11.32q12.1).Among the nine fetuses,including seven from five pedigrees reported in the literature retrieved and two from the two pedigrees we reported,seven showed abnormal soft markers or structures in ultrasound and three of the seven pedigrees had high risk of fetal trisomy 18.Conclusions Ultrasound screening is highly sensitive in detecting rec(18) fetuses,yet the association between ultrasound features and fetal karyotypes is not clear.The combination of multiple genetic analysis methods,including karyotype analysis,CMA and FISH,may be conducive to clarifying the types and sources of complex derived chromosomes.

Objective To study the expressions of HnRNPA1 in patients with recurrent or metastatic hepatocellular carcinoma (HCC) after liver transplantation.Methods The expressions of HnRNPA1 protein in pericarcinoma and normal liver tissues were detected using Western blot analysis in 16 patients with HCC.Immunohistochemical analysis was done in 141 patients with HCC.All these patients underwent liver transplantation.The relationship between the expressions of HnRNPA1 with recurrent and metastatic HCC were analyzed.Results The positive expression rate of HnRNPA1 protein in the HCC tissues (75.0％,12/16) was significantly higher than that in the pericarcinoma tissues (18.8％,3/16) (P ＜ 0.01).The expression of HnRNPA1 was positively correlated with tumor size,TNM type,vascular invasion and tumor encapsulation (P ＜ 0.01).Tumor recurrence in the HnRNPA1 high expression group was significantly higher than that in the HnRNPA1 low expression group (x2 =15.577,P ＜ 0.01).The survival rate was significantly lower in the high HnRNPA1 expression than that in the low expression group (x2 =6.309,P ＜0.05).Conclusion The expression of HnRNPA1 protein was a marker which predicted HCC recurrence or metastasis in patients after liver transplantation.