Unexpected detection of Duchenne muscular dystrophy gene mutation by array comparative genomic hybridization in prenatal diagnosis: a retrospective analysis / 中华围产医学杂志

Objective:To explore the accuracy of array comparative genomic hybridization(aCGH) in the unexpected detection of Duchenne muscular dystrophy ( DMD) gene duplication/deletion in prenatal diagnosis. Methods:A retrospective analysis was performed on 31 cases with DMD gene duplication/deletion detected by aCGH among 5 025 prenatal diagnosis samples without family history of DMD in Henan Provincial People's Hospital from July 2018 to December 2019. The multiplex ligation-dependent probe amplification (MLPA) method was used to verify the above results. The American College of Medical Genetics and Genomics (ACMG) guideline was referred for pathogenicity analysis of the detected duplicates/deletions. Descriptive analysis was adopted in analysis. Results:The total unexpected DMD gene duplication/deletion rate was 0.62% (31/5 025), among which 25 cases were with microduplication/microdeletion ≤ 200 kb and six were >200 kb; there were 24 cases of deletion, seven cases of duplication; exon or intron duplication/deletion were accounted for 19 and 12 cases, respectively. According to the five classification standards of ACMG guideline, there were 17 cases with pathogenic variants and 14 cases with uncertain pathogenicity/likely benign variants. Of the 19 with exon mutations, 17 cases were DMD intragenic variants, and two cases involved variants in and outside DMD gene, which were verified by MLPA whose results were all positive. Conclusions:The duplication/deletion of exon region of DMD gene detected by aCGH technique is accurate and reliable, which plays an important role in the diagnosis of DMD. For these cases involved both internal and external regions of DMD gene, aCGH can identify the upstream and downstream breaking points of DMD gene, thus providing the basis for ACMG grading.

Clinical study of Xuebijing Injection in treatment of capillary leak syndrome / 国际检验医学杂志

Objective To study the clinical application effect of Xuebijing Injection in the treatment of capillary leak syndrome.Methods One hundred patients with capillary leak syndrome in our hospital from January 2013 to April 2016 were selected and randomly divided into the control group and observation group according to the ID number of admission.The two groups were given the Western medicine routine therapy of capillary leak syndrome.The observation group was simultaneously given Xuebijing Injection.The APACHE II score,Mashall score and hemodynamics were compared between the two groups.The disease condition change was dynamically observed.Results The APACHE II score and Mashall score before and after treatment in the observation group were significantly suprior to those in the control group(P<0.05);the peripheral blood circulating endothelial cells (CEC) number at admission in the two groups were higher than the normal level,the CEC content after taking the treatment was gradually decreased (P<0.05),but the CEC content decrease in the observation group was less than that of the control group (P<0.01);were observed before and after the treatment group APACHE II and Mashall score was significantly better than the control group (P<0.05);but the CEC content in the observation group was significnatly superior to the control group(P<0.01);the expressions of vascular endothelial growth factor (VEGF),TNF alpha and interleukin-6 (IL-6) in the observation group were better than those in the control group (P<0.05).Conclusion Using Xuebijing Injection in treating capillary leak syndrome can effectively protect the vascular endothelial cells,improves the living quality of patients and can be popularized and applied in clinical treatment.

Cloning of keratinocyte growth factor 2 gene (KGF2) and its transformation to Brassica napus L / 生物工程学报

Recently, more research about the plant bioreactor expressing genes encoding human proteins was reported. In the present study, the cDNA of the human gene keratinocyte growth factor 2 (KGF2) was replaced with plant preferred codons by PCR, and the modified full-length cDNA was cloned into the plant expression vector pCAMBIA-YO containing the oil-body promoter. The fusion construct pCAMBIA-YO-KGF2 was transformed into Brassica napus by Agrobacterium tumefacien-mediated cotyledon transformation method. The transgenic seedlings were identified by PCR, Southern and western blot analysis all showed that KGF2 gene was successfully expressed in in transgenic Brassica napus.

Development of genetic transformation system and transformation of active fibroblast growth factor (aFGF) in Carthamus tinctorius / 中国中药杂志

<p><b>OBJECTIVE</b>To establish genetic transformation system of active fibroblast growth factor (aFGF) in Carthamus tinctorius.</p><p><b>METHOD</b>The culture condition was optimized by orthogonal experiment design with cotyledon of C. tinctorius as the explant. The aFGF was transferred into safflower through Agrobacterium-mediated transformation and screened under different concentrations of antibiotics, and then PCR was identified.</p><p><b>RESULT</b>It confirmed the optimal differentiation medium: MS + BA 1.0 mg x L(-1) + NAA 0.2 mg x L(-1), the optimal root medium: 1/4 MS + NAA 2.0 mg x L(-1) + IAA0.5 mg x L(-1). The bacteriostatic effect of the three antibiotics showed slight difference. From them Tim was selected with the concentration of 400 mg x L(-1). It showed the bacteriostatic effect and promoted also differentiation. The selective concentration of hyg was confirmed to be 6 mg x L(-1). The eight transformed plants were identified, the positive rate was 25%.</p><p><b>CONCLUSION</b>It was determined the best hormones and the ratios for the differentiation and rooting of the safflower by organogenesis. It was identified the optimal concentration of inhibitory antibiotics and selection antibiotics. The aFGF gene was cloned in a part of plant by PCR analysis. It is shown that the aFGF gene has been integrated into safflower genome.</p>

Application of the self excision Cre/lox system in plants / 生物工程学报

Marker-free plants have been public concern. Co-transformation and site-specific recombination system are more important methods in self-gene excision. We reviewed the Cre/lox site-specific system and its applications in plants, also, we discussed perspectives of the system in according with our experience.