[Screening for hotspot mutations associated with genetic hearing impairment in pregnant women and subsequent prenatal diagnosis in high risk pregnancies].

Objective: To screen for hotspot gene mutations associated with genetic deafness in Chinese pregnant women, and to perform risk assessment and prenatal diagnosis in high-risk families. Methods: Between November 2012 and October 2017, 26 117 pregnant women were screened by molecular hybridization microarray for 9 hot-spot mutations in 4 hereditary deafness related genes (GJB2 c. 35 del G, c. 176_191 del 16 bp, c. 235 del G, c. 299_300 del AT, GJB3 c. 538 C>T, SLC26A4 c. 2168 A>G, IVS 7-2 A>G, mitochondrial DNA 12S rRNA m. 1494 C>T, m. 1555 A>G). Genotype analysis was carried out in husbands of women carrying mutations, and prenatal diagnosis was carried out in the fetuses with high risk of deafness. Results: Among all women tested, 1 208(4.63%) were carriers of genetic deafness mutations, 7 with hearing impairment were affected by homozygous or compound heterozygous mutations, 51 were mitochondrial gene mutation carriers, 103 were carriers of GJB3 c. 538 C>T heterozygous mutation, 1 026 were carriers of GJB2 or SLC26A4 heterozygous mutations, and 21 carried heterozygous mutations in two genes simultaneously. In 394 families, the husbands accepted gene sequence testing, and 27 in which were determined as carriers of mutations in identical genes as their wives. Among which, 18 families received prenatal diagnosis, and 5 fetuses were diagnosed as hereditary deafness. In 9 families who did not receive prenatal diagnosis, 1 neonate was diagnosed as compound heterozygote after delivery. Conclusion: In order to prevent birth defects with congenital hearing problems, it is effective to provide screening for hotspot mutations in pregnant women and to perform prenatal diagnosis on high risk pregnancies.

[Application of copy number variation analysis based on next-generation sequencing in the genetic analysis of missed abortion chorionic villi].

OBJECTIVE: To investigate the value of copy number variation analysis based on next-generation sequencing (NGS-CNVA) in the genetic analysis of missed abortion chorionic villi. METHODS: From August 2012 to May 2014, chorionic villi from 74 cases of missed abortion at 6-13 gestational weeks in Haidian Maternal and Child Health Hospital were collected and analyzed by karyotype analysis and NGS-CNVA. The results of the two methods were compared. RESULTS: (1) Karyotype analysis was carried out for the villi from the 74 missed abortion patients. Thirty cases were euploid, 26 cases were aneuploid, while 18 cases had structural abnormalities. The resolution of the karyotyping was 320 bands and the average report time was 22 days. (2) All of the 74 samples obtained NGS-CNVA results and the report time was 7-10 days. (3) The NGS-CNVA results of 56 cases were consistent with karyotype. Among them, 28 cases (28/56, 50%) had no copy number variants (CNV), and 19 cases (19/56, 34%)had CNV between 1 Mb and 10 Mb. 9 cases (9/56,16%) had CNV ≥10 Mb found by NGS-CNVA, but not found by karyotyping. (4) According to the results of NGS-CNVA, karyotype were reviewed. The reviewed results found 7 cases with CNV<10 Mb and 3 cases with CNV≥10 Mb in 30 cases which got normal karyotype results at the first analysis. (5) Among the 18 cases of structural abnormalities, 6 cases were Robertsonian translocation. Sequencing technology could confirm the specific area of chromosome deletion/duplication in 8 cases, but could not locate them. CONCLUSIONS: NGS-CNVA has lower failure rate, higher resolution, lower specimen requirement and shorter report time than karyotype analysis when used for the genetic analysis of missed chorionic villi. NGS-CNVA could be a useful genetic analysis method for the missed abortion villi.

[Experimental study on preventive and therapeutic effect of bushen huoxue recipe on autoimmune premature ovarian failure model].

OBJECTIVE: To investigate the mechanisms of Bushen Huoxue Recipe (BSHXR), a Chinese herbal medicinal preparation for tonifying Kidney and invigorating blood circulation, to prevent and treat autoimmune premature ovarian failure (POF) model. METHODS: Female 8-10 weeks old BAL B/c mice were immunized by intracutaneously injecting porcine ovum zona pellucida (ZP), isolated by immuno-chromatography, in multiple points of two hind footpads to establish the POF model and treated with BSHXR started from early stage of immunization (prevented group) or after 3 times of injection (treated group). Changes in vaginal smears, serum estradiol (E2), antibody titer against ZP, response of splenic lymphocyte to ZP stimulation of different concentrations, and numbers of ovarian follicles and corpus luteum were analyzed. RESULTS: Serum E2 level in the prevented and treated mice was all higher than that in the non-treated immunized model mice (the control group), P < 0.01 and P < 0.05 respectively. But the anti-ZP titer lowered significantly after BSHXR administration, as compared with that in the control group. Level of antibodies in the treated group was lower than that of the control, and it was also lower in the prevented group than that in the control. The histo-morphological examination showed that the developing follicles and corpus luteum after BSHXR medication in both prevented and treated group increased significantly as compared with that of the control group (P < 0.05 and P < 0.01 respectively). Splenic lymphocyte in the immunized model showed a higher antigen-specific proliferation reaction than that in non-immunized animal, and the reaction was ameliorated by BSHXR medication. CONCLUSION: BSHXR could recover part of the ovarian function in POF mice mainly through inhibiting specific immune injury to revive the remnant follicles in ovary. The preventive effect of BSHXR was superior to the therapeutic effect of it.

[Cloning and sequencing of a DNA fragment from Plasmodium falciparum FCC1/HN isolate].

A specific DNA fragment isolated from Plasmodium falciparum FCC1/HN isolate has been cloned in Bam H1 site of pUC18 and first partially sequenced by Sanger's method. The results show: G+C percent of DNA sequence is 26.8%, and the cloned DNA has many restriction sites which are conventional for subcloning. The authors suggest that this clone and sequence may be used as a guide for developing a DNA probe or PCR primers.