AIM To investigate whether tubeimosides induce apoptosis in CNE-2Z. METHODS Growth inhibition by tubeimosides was measured using MTT assay. The effect of tubeimosides on apoptosis induction of CNE-2Z cell line was studied by the fluorescent microscopy, electronic microscopy, DNA agarose gel electrophoresis, flow cytometry analysis. Western blotting was performed for detecting the apoptosis-related gene. RESULTS The growth of CNE-2Z cells was inhibited obviously by tubeimosides. CNE-2Z cells showed typical apoptotic features observed by fluorescent microscopy, electronic microscopy. DNA fragmentation into multiples of low molecular weight DNA(180～200 bp) was detected by agarose gel electrophoresis of DNA; Sub-G 1 peak was found by flow cytometry analysis, and the apoptosis index was 58 0%. The expression of bcl-2, inhibitor of apoptosis, decreased apparently; while that of bax, inducer of apoptosis, increased notably at 1, 3, 5 h after the addition of tubeimosides. The cleavage of caspase-3 was noticed at 1, 3, 5 h after the treatment of tubeimosides. CONCLUSION Tubeimosides could induce the apoptosis of CNE-2Z cells. The induction of apoptotic by tubeimosides was closely associated with bcl-2, bax and caspase-3.

Objective To investigate the value of prenatal diagnosis in identifying the etiology and predicting the prognosis of fetal pleural effusion (FPE).Methods Forty-two cases of FPE were recruited in this study from January 2012 to September 2016.Ultrasound scan and genetic tests were performed on all fetuses.Seven fetuses with severe FPE were given pleurocentesis.Pregnancy outcomes of all the fetuses were followed up.Results FPE was commonly accompanied with other abnormalities,such as ascites,hydrops,hydramnion,hygroma colli,abnormal posturing,joint contractures,arrhythmia and micromandible.Chromosomal abnormality was detected in 11 fetuses (26.2％),of which ten were further confirmed by karyotype analysis,including six with 45,X,three trisomy 21 and one trisomy 18,and one was detected with a 9.83 Mb uniparental disomy (UPD) located at 12q24.21q24.31 by gene chip.One fetus was diagnosed with--SEA/--SEA thalassemia.All of the 12 families decided to terminate the pregnancies after genetic counseling.Among the other 30 fetuses,seven with severe FPE and normal karyotype underwent pleurocentesis.Five of the seven cases were with favorable outcomes,one with progressive hydrops was aborted and one neonate with severe hydrops died after birth.Spontaneous regression of FPE with good outcome was found in two cases.Parents of the other 21 fetuses chose to terminate the pregnancies.Conclusions Prenatal diagnosis is important to identify the etiology and predict the outcome of FPE.Chromosomal abnormality is a relatively common cause of FPE,and 45,X and trisomy 21 are the most common abnormalities.Intrauterine intervention is beneficial for FPE without chromosomal or other definite genetic abnormalities.Genetic test may be of great value for pregnant counseling.

Non-immune factors are the most common cause of hydrops fetalis, which may manifest as abnormal accumulation of fluid in the body cavities. Hydrops fetalis is a complex condition with a high mortality rate. The prognosis is depending on the underlying disease and is particularly poor in those with chromosomal aberrations or monogenic disorders. In this paper, advance in the research on non-immune hydrops fetalis is reviewed.

Objective: To carry out genetic testing for a family with two pregnancies affected with hydrops fetalis and dilated cardiomyopathy (DCM) of the fetus. Methods: DNA was extracted from fetal tissue as well as peripheral blood samples from the couple. Single nucleotide polymorphism array (SNP array) and next-generation sequencing (NGS) were carried out to screen potential mutation. Suspected mutation was validated with PCR and Sanger sequencing. Results: The manifestation of fetal echocardiography was consistent with DCM. No obvious abnormality was found by SNP array analysis. A hemizygous c. 481G>A (p.G161R) mutation of the TAZ gene was detected in the male fetus by NGS and confirmed by Sanger sequencing. The mutation was inherited from his mother. Conclusion Barth syndrome due to the c. 481G>A mutation of the TAZ gene probably underlies the recurrent hydrops fetalis and fetal DCM in this family.

OBJECTIVE: To explore the genetic etiology for a pedigree affected with progressive familial intrahepatic cholestasis (PFIC). METHODS: Target sequence capture and next generation sequencing (NGS) were applied for the proband. PCR and Sanger sequencing were used to verify the suspected mutation in his sister with similar symptoms and his parents. RESULTS: The proband and his sister manifested after birth with symptoms including jaundice, pruritus and developmental retardation. NGS has identified compound heterozygous mutations of ABCB11 gene, which encodes bile salt export pump protein (BSEP), namely c.2494C>T (p.Arg832Cys) and c.3223C>T (p.Gln1075*), in the proband, which were inherited from his father and mother respectively. His sister carried the same compound mutations. CONCLUSION Based on the phenotype and genetic testing, the patients were diagnosed as PFIC2 caused by mutation of the ABCB11 gene. The c.3223C>T is a novel nonsense mutation which may cause premature termination of translation. Above results have enriched the spectrum of ABCB11 mutations and provided new evidence for the molecular basis of PFIC, which also facilitated genetic counseling for this pedigree.
ATP Binding Cassette Transporter, Subfamily B, Member 11 ; genetics ; ATP-Binding Cassette Transporters ; Cholestasis, Intrahepatic ; genetics ; Female ; Genetic Testing ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

OBJECTIVE: To explore the clinical features and genetic diagnosis of two cases with rare diseases and X chromosome abnormalities. METHODS: Multiple ligation-dependent probe amplification (MLPA) and karyotype analysis were carried out on an 8-year-old girl who was diagnosed with Duchenne muscular dystrophy. Karyotype analysis and PCR assay for SRY and AZF genes were carried out for a-2-month-old male infant with short penis. RESULTS: The girl, who featured short stature and cubitus valgus, was diagnosed as Turner syndrome with a karyotype of 46,X,i(Xq). The male infant was detected with a karyotype of 45,X, with presence of SRY gene but absence of AZF gene. CONCLUSION Both cases may be associated with abnormalities of X chromosome. Genetic testing can facilitate early diagnosis and clinical intervention for such patients.
Chromosomes, Human, X ; Humans ; Infant ; Karyotyping ; Male ; Muscular Dystrophy, Duchenne ; genetics ; Rare Diseases ; Turner Syndrome ; genetics

OBJECTIVE: To carry out genetic testing for a family with two pregnancies affected with hydrops fetalis and dilated cardiomyopathy (DCM) of the fetus. METHODS: DNA was extracted from fetal tissue as well as peripheral blood samples from the couple. Single nucleotide polymorphism array (SNP array) and next-generation sequencing (NGS) were carried out to screen potential mutation. Suspected mutation was validated with PCR and Sanger sequencing. RESULTS: The manifestation of fetal echocardiography was consistent with DCM. No obvious abnormality was found by SNP array analysis. A hemizygous c.481G>A (p.G161R) mutation of the TAZ gene was detected in the male fetus by NGS and confirmed by Sanger sequencing. The mutation was inherited from his mother. CONCLUSION Barth syndrome due to the c.481G>A mutation of the TAZ gene probably underlies the recurrent hydrops fetalis and fetal DCM in this family.
Barth Syndrome ; genetics ; Cardiomyopathy, Dilated ; genetics ; Echocardiography ; Female ; Genetic Testing ; High-Throughput Nucleotide Sequencing ; Humans ; Hydrops Fetalis ; genetics ; Male ; Mutation ; Pregnancy ; Transcription Factors ; genetics

Objective To evaluate 3D printing abdominal aortic aneurysm model in analysis of clinical teaching effect for standardized resident doctors in vascular surgery department. Methods 48 resi-dents in vascular surgery department in our hospital from December 2016 to September 2017 were seleeected and randomly divided into control group and the experimental group. The traditional vascular surgical anatomy atlas, ultrasound, CT abdominal blood vessel 3D reconstruction, digital subtraction, video and so on were used by 24 residents in the control group, while in the experimental group, on the basis of the traditional teaching, abdominal aortic aneurysm model of 3D printing, true aortic coated stent delivery system were increased. After the teaching, the theory of evaluation (abdominal vascular anatomical features, morphological characteristics and classification of AAA, measurement of various parameters and key points of operation in the EVAR) and satisfaction questionnaire were adopted to evaluate the effect of two kinds of teaching methods. SPSS 19.0 was used to conduct t test on two groups of physician evaluation data. Results The results of theoretical assessment showed that there was no significant difference between the control group and the experimental group in the abdominal vascular anatomical features, the morphological features and the classification of the experimental group (P>0.05) in the examination of the common AAA cases and the complicated AAA cases. However, the experimental group was higher than the control group in the mea-surement of the parameters of EVAR, and the score of the operation points and the total score, and the difference was statistically significant (t=2.283, t=2.263, P<0.05). The results of the questionnaire showed that the students' satisfaction scores on the teaching satisfaction of the normal and complex AAA cases were better than those of the control group, and the difference was statistically significant (P<0.05). Conclusion The 3D printing model can increase the understanding and mastery of the anatomy and treat-ment of abdominal aortic aneurysm and improve its learning enthusiasm for vascular surgery. We should make full use of the advantages of 3D printing technology on the basis of retaining the advantages of tradi-tional teaching methods and means, and further enhance the teaching effect.

Objective: The phenotype and genetics of three patients with autosomal recessive polycystic kidney disease (ARPKD) at childhood, teenage and advanced age were analyzed. Methods: Next generation sequencing (NGS) was applied to all the probands. PCR and Sanger sequencing were used to verify the suspicious gene variants screened by NGS in the probands and their family members, and one of the family got prenatal diagnosis. Results: Through NGS, PCR and Sanger sequencing, the 5-yr proband in pedigree 1 was shown to carry compound heterozygous variants of c. 5935G>A(p.G1979R) and c. 5428G>T(p.E1810X) of PKHD1, originated from his parents; In pedigree 2, the 17-ys proband was detected with c. 5512T>C(p.Y1838H) and c. 5935G>A(p.G1979R) variants of PKHD1 orginated from her parents, and her mother also got prenatal diagnosis during the second trimester; In pedigree 3, the 70-ys female proband was found with variants c. 11314C>T (p.R3772X) and c. 3860T>G (p.V1287G) of PKHD1. Conclusion The three pedigrees were diagnosed as ARPKD caused by PKHD1 variants. Five types of variants were detected, c. 5935G>A and c. 11314C>T were the known pathogenic variants, while c. 5512T>C, c. 5428G>T and c. 3860T>G were not reported previously. Considering the complexity of the genetics and phenotypes of the cystic renal diseases, genetic diagnosis is crucial to give accurate etiological diagnosis, which may benefit the clinic management.