Significance and case analysis of FMR1 mutation screening during early and middle pregnancy / 中华医学遗传学杂志

OBJECTIVE@#To screen for mutations of fragile X mental retardation 1 (FMR1) gene during early and middle pregnancy and provide prenatal diagnosis for those carrying high-risk CGG trinucleotide expansions.@*METHODS@#Peripheral blood samples of 2316 pregnant women at 12 to 21(+6) gestational weeks were collected for the extraction of genomic DNA. CGG repeats of the FMR1 gene were detected by fluorescence PCR and capillary electrophoresis. Genetic counseling and prenatal diagnosis were provided for 3 women carrying the premutations.@*RESULTS@#The carrier rate of CGG repeats of the FMR1 gene was 1 in 178 for the intermediate type and 1 in 772 for the premutation types. The highest frequency allele of CGG was 29 repeats, which accounted for 49.29%, followed by 30 repeats (28.56%) and 36 repeats (8.83%). In case 1, the fetus had a karyotype of 45,X, in addition with premutation type of CGG expansion of the FMR1 gene. Following genetic counseling, the couple chose to terminate the pregnancy through induced labor. The numbers of CGG repeats were respectively 70/- and 29/30 for the husband and wife. In case 2, amniocentesis was performed at 20 weeks of gestation. The number of CGG repeats of the FMR1 gene was 29/-. No abnormality was found in the fetal karyotype and chromosomal copy number variations. The couple chose to continue with the pregnancy. Case 3 refused prenatal diagnosis after genetic counseling and gave birth to a girl at full term, who had a birth weight of 2440 g and no obvious abnormality found during follow-up.@*CONCLUSION@#Pregnant women should be screened for FMR1 gene mutations during early and middle pregnancy, and those with high-risk CGG expansions should undergo prenatal diagnosis, genetic counseling and family study.

Primary cilia mediate Klf2-dependant Notch activation in regenerating heart

Unlike adult mammalian heart, zebrafish heart has a remarkable capacity to regenerate after injury. Previous study has shown Notch signaling activation in the endocardium is essential for regeneration of the myocardium and this activation is mediated by hemodynamic alteration after injury, however, the molecular mechanism has not been fully explored. In this study we demonstrated that blood flow change could be perceived and transmitted in a primary cilia dependent manner to control the hemodynamic responsive klf2 gene expression and subsequent activation of Notch signaling in the endocardium. First we showed that both homologues of human gene KLF2 in zebrafish, klf2a and klf2b, could respond to hemodynamic alteration and both were required for Notch signaling activation and heart regeneration. Further experiments indicated that the upregulation of klf2 gene expression was mediated by endocardial primary cilia. Overall, our findings reveal a novel aspect of mechanical shear stress signal in activating Notch pathway and regulating cardiac regeneration.

Primary cilia mediate Klf2-dependant Notch activation in regenerating heart

Unlike adult mammalian heart, zebrafish heart has a remarkable capacity to regenerate after injury. Previous study has shown Notch signaling activation in the endocardium is essential for regeneration of the myocardium and this activation is mediated by hemodynamic alteration after injury, however, the molecular mechanism has not been fully explored. In this study we demonstrated that blood flow change could be perceived and transmitted in a primary cilia dependent manner to control the hemodynamic responsive klf2 gene expression and subsequent activation of Notch signaling in the endocardium. First we showed that both homologues of human gene KLF2 in zebrafish, klf2a and klf2b, could respond to hemodynamic alteration and both were required for Notch signaling activation and heart regeneration. Further experiments indicated that the upregulation of klf2 gene expression was mediated by endocardial primary cilia. Overall, our findings reveal a novel aspect of mechanical shear stress signal in activating Notch pathway and regulating cardiac regeneration.

Identification of a novel CRYGC mutation in a pedigree affected with congenital cataracts / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a Chinese pedigree affected with congenital cataracts.@*METHODS@#Clinical data and peripheral blood samples were collected for the pedigree. Following extraction of genomic DNA, whole exome sequencing was carried out to detect genetic variants. Candidate variants were verified by familial co-segregation analysis and Sanger sequencing. Bioinformatics analysis was carried out to predict the function of mutant genes.@*RESULTS@#By comparing variants identified among affected and unaffected individuals, a heterozygous variant, c.110 G>C (p.R37P), was identified in exon 2 of the CRYGC gene among all patients, which also matched the criteria for potential disease-causing mutations. The result was confirmed by Sanger sequencing.@*CONCLUSION@#The c.110G>C variant of the CRYGC gene probably underlay the congenital cataracts in this pedigree.

Screening for differentially expressed proteins in patients with liver cirrhosis complicated by portal vein thrombosis / 临床肝胆病杂志

Objective To investigate the differentially expressed serum proteins in patients with liver cirrhosis complicated by portal vein thrombosis (PVT) . Methods Serum samples were collected from 45 patients with liver cirrhosis who were hospitalized in Infectious Disease Center, The First Affiliated Hospital of Xinjiang Medical University, from November 2015 to November 2016, and among these patients, 22 had PVT and 23 had no PVT. Isobaric tags for relative and absolute quantitation (i TRAQ) combined with chromatography and mass spectrometry were used to screen out the differentially expressed serum proteins, and a bioinformatics analysis was performed for the differentially expressed proteins. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups, and the chi-square test was used for comparison of categorical data between groups. The Fisher's exact test was used to compare the distribution of GO terms or KEGG pathways in the target protein set or total protein set, in order to evaluate the significance level of protein enrichment of a GO term or KEGG pathway. Results A total of 800 proteins were screened out, among which 86 were differentially expressed, including 32 upregulated proteins (ratio ≥1. 2, P ＜ 0. 05) and 54 downregulated proteins (ratio ≤0. 83, P ＜ 0. 05) . Among these proteins, 14 were associated with cellular component, 22 were involved in biochemical processes, and 10 were associated with molecular function. The KEGG analysis showed that there were significant differences in 18 proteins in 5 metabolic and signaling pathways between the liver cirrhosis-PVT group and liver cirrhosis group. These 5 metabolic and signaling pathways were associated with fat digestion and absorption, platelet activation, metabolism of glyoxylic acid and dicarboxylic acid, osteoclast differentiation, and axon guidance. Conclusion i TRAQ combined with chromatography and mass spectrometry can effectively screen out the differentially expressed serum proteins, among which GP5, FGA, and FGG may be potential biological markers for PVT in patients with liver cirrhosis and are worthy of further research.

Effects of electroacupuncture at "Weizhong" (BL 40) on regeneration and morphology in rats with bupivacaine-induced multifidus muscle injury / 中国针灸

<p><b>OBJECTIVE</b>To observe the intervention effect of electroacupuncture (EA) at "Weizhong" (BL 40) on rats with bupivacaine-induced multifidus muscle injury, so as to explore the action mechanism.</p><p><b>METHODS</b>A total of 72 rats were randomly divided into a control group, a model group, a Weizhong group and a Shenshu group, 18 rats in each group. Each group was again randomly divided into a 4-day subgroup, a 7-day subgroup and a 14-day subgroup, 6 rats in each subgroup. Rats in the model group, Weizhong group and Shenshu group were treated with intramuscular injection of 0.5% bupivacaine (BPVC) to establish the model of multifidus muscle injury. Rats in the Weizhong group and Shenshu group were treated with EA at "Weizhong" (BL 40) and "Shenshu" (BL 23), 20 min per treatment, once a day. Each subgroup was treated for 4 days, 7 days and 14 days respectively. Rats in the control group and model group were treated with immobilization. The morphology and cross sectional area (CSA) changes of multifidus with HE and Masson staining at different time points were observed; the expression of insulin like growth factor 1 (IGF-1) and myogenic differentiation antigen (MyoD) was measured by immunohistochemical method.</p><p><b>RESULTS</b>After the modeling, there were significant morphology changes of multifidus at different time points, which was not fully recovered after 14 days. The morphological observation in the Weizhong group and Shenshu group was superior to that in the model group. At 7th day, the CSA in the Weizhong group was higher than that in the model group (P < 0.05); at 14th day, the CSA in the Weizhong group and Shenshu group was higher than that in the model group (P < 0.01, P < 0.05). At 4th day and 7th day, the expression of IGF-1 in the model group was higher than that in the control group (both P < 0.01); at 4th day, that in the Weizhong group was higher than that in the model group (P < 0.01), and that in the Weizhong group was higher than that in the Shenshu group (P < 0.05), and that in the Shenshu group was as higher than that in the model group (P < 0.05); at 14th day, that in the Shenshu group was higher than that in the model and Weizhong group (P < 0.01). At 4th day, the expression of MyoD in the Weizhong group and Shenshu group was higher than that in the model group (P < 0.01), which was more significant in the Weizhong group (P < 0.01).</p><p><b>CONCLUSION</b>Electroacupuncture at "Weizhong" (BL 40) and "Shenshu" (BL 23) can both promote the regeneration of multifidus muscle injury. EA at "Weizhong" (BL 40) has a better effect at early phase, which may be related to the up-regulation of IGF-1 and MyoD and the completion of the proliferation of myoblast in advance.</p>

Screening of mutations of deafness-related genes in women of child-bearing age from Shijiazhuang area / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To screen for mutations of deafness-related genes among ethic Chinese women of child-bearing age.</p><p><b>METHODS</b>In 324 women, 9 mutational sites in 4 deafness-related genes (SLC26A4, GJB3, GJB2 and mtDNA 12s rRNA) were screened using a gene chip.</p><p><b>RESULTS</b>Twenty women (6.17%) have carried mutations. These included 11 (3.40%) carrying a GJB2 gene mutation, 7 (2.16%) carrying a SLC26A4 gene mutation, 1 (0.31%) simultaneously carrying GJB3 and GJB2 gene mutations, and 1 (0.31%) carrying a mtDNA 12s rRNA gene mutation.</p><p><b>CONCLUSION</b>Women of child-bearing age have a high rate for carrying mutations of common deafness-related genes, among which 235delC in GJB2 was most common. Prenatal screening of couples with normal hearing is an effective way to prevent birth of affected children.</p>

Effects of Electroacupuncture on Morphology Repairment and Basic Fibroblast Growth Factor/Extracellular Signal-regulated Kinase Signal Pathway of Rabbits with Acute Lumbar Muscle Contusion / 中国康复理论与实践

@#Objective To observe the effect of electroacupuncture on basic fibroblast growth factor (bFGF), Desmin and extracellular signal-regulated kinase (ERK1/2) expression of rabbits with acute lumbar muscle contusion, and discuss the possible mechanism of muscle tissue regeneration and repair. Methods 40 male New Zealand rabbits were randomly divided into blank group (BG, n=10), model group (MG, n=10), Weizhong (BL40) electroacupuncture group (WG, n=10) and local electroacupuncture group (LG, n=10). Lumbar muscle injury was established with blunt trauma, and the rabbits were assessed with Appearance Score. The WG and LG accepted electroacupuncture for 2 weeks after modeling. The lumbar muscle of each animal was stained with HE to observe pathological changes, and immunohistochemical staining was used to observe bFGF expression, and Western blotting was used to detect the expression of Desmin and ERK1/2 protein.Results Appearance Score was more in the MG, WG and LG than in the BG (P<0.01). Histological score ranked from more to less as MG,LG, WG and BG (P<0.05). The expression of bFGF ranked as LG, WG, MG and BG (P<0.01), Desmin ranked as WG=LG and BG=MG (P<0.05), ERK1/2 as LG=WG, MG and BG (P<0.05). Conclusion Electroacupuncture could promote the regeneration and repair of tissue after acute lumbar muscle contusion, which may be related to the up-regulation of bFGF/ERK signaling pathway.

New multiplex-PCR assay for detection of deletions of DMD gene in Chinese DMD patients / 中华检验医学杂志

Objective To establish a new multiplex-PCR assay to improve the detection rate of mutations in the DMD gene in Chinese patients. Methods A retrospective review of DMD deletion spectrum of 355 DMD patients with deletions all over the gene was performed. All deletions were confirmed by " one-step approach" diagnostic procedure and MLPA analysis. The exons with high frequency of mutations were identified to constitute the amplification system and the PCR conditions were optimized. Results Two new multiplex-PCR assays were established. Assay one was used to detect 10 exons including exon 5, 8, 17, 44, 45, 47, 49, 50, 51 and 52 of DMD gene, in two PCR sets. The theoretical detection rate would be 92% (326/355). Assay two was used to detect 5 exons including exon 12, 19, 35, 43 and 54, which could be used to screen additional 5% (17/355) deletion cases. The method was validated in other 22 DMD patients. Multiplex-PCR results were completely identical to the MLPA results in all 22 DMD patients. Conclusions The two multiplex-PCR assays were established based on the analysis of 355 Chinese DMD patients with gene deletions. It is believed that the new approach would be more applicable for deletion detection on the Chinese DMD patients since the DMD cases involved were from the whole country.

A Simplified Approach for Detecting Homologous Deletion of SMN1 Genes in Spinal Muacular Atrophy / 医学研究杂志

Objective To develop a rapid,reliable and convenient approach for diagnosing the homozygous deletion of SMN1 gene.Methods SMN1 gene was amplified specifically with double allele-specific PCR(AS-PCR).Meanwhile,one inrelevant gene was amplified as internal control by PAGE and agarose gel electrophoresis analysis to determine whether the sick children were with homozygous deletion of SMN1 genes.Results The homozygous deletion of exon7 in SMN1 gene was identified by agarose gel electrophoresis or PAGE accurately.Conclusion Compared to PCR-RFLP and DHPLC used in the past,this approach can diagnose homozygous deletion of SMA much more accurate,easier and more convenient without completed following analyses.

A Novel Mutation of ADAR Gene Identified in a Chinese Pedigree with Dyschromatosis Symmetrical Hereditaria / 医学研究杂志

Objective To discover the mutation of ADAR gene in a pedigree with dyschromatosis symmetrical hereditaria(DSH). Methods We investigated this family and collected blood samples of the individuals in this family. Mutation screening was carried out by PCR and direct sequencing. The allele specific primer was designed for the mutation point, and allele-specific PCR was carried out on the patients, normal family members and 40 normal individuals. Results A single nucleotide deletion (c.1642 delC) was identified in exon3 of ADAR gene in the patients of this family. This mutation was not detected in the normal family members and in any of the control individuals. Conclusion This single nucleotide deletion was responsible for the disease in the family.

Construction of HNP-3 Mature Peptide Bait Plasmid of Yeast Two-hybrid System and Detecting Its Self-activating and Toxic Effect / 航天医学与医学工程

Objective To construct the bait plasmid of HNP-3 mature peptide in yeast two-hybrid system and examine whether the recombinant bait plasmid has self-activating and toxicity effect.Methods Using RT-PCR technique,the cDNA fragments of HNP-3 mature peptide gene were amplified from the extracted RNA in cultured HL-60 cells.The fragment was firstly cloned into pBluescript-SK-II vector,confirmed by sequencing,then sub-cloned into the bait plasmid pGBKT7 and identified with PCR and sequence analysis techniques.The recombinant plasmid was introduced into the yeast cell AH109,and its self-activating and toxicity effect was tested by auxotrophic selective culture.Results DNA sequencing indicated that the inserted fragment in pBluescript-SK-II vector was HNP-3 mature peptide gene sequence,and the sub-cloned recombinant pGBKT7-HNP-3 was no mismatch.The recombinant bait plasmid didn't have self-activating effect and did not show toxicity to yeast AH109 cell.Conclusion The bait plasmid of HNP-3 mature peptide was constructed successfully.This was helpful for investigating the proteins interacting with HNP-3 mature peptide by yeast two-hybrid technique.