AIM: To study the effect of adrenomedullin (ADM) on L-type calcium currents (I Ca,L) and its signaling transduction process in guinea-pig ventricular myocytes. METHODS: By using whole-cell patch clamp technique, the effect of ADM(1-100 nmol?L -1) on I Ca,Lmyocyte. Furthermore, the possible signaling transduction process of ADM on I Ca,L was investigated by observing the effects of ADM 22-52 (a specific ADM-receptor antagonist,100 nmol?L -1)+ADM(100 nmol?L -1), H-89 (a specific protein kinase A inhibitor, 10 ?mol?L -1) + ADM(100 nmol?L -1), PKC 19-36 (a specific protein kinase C inhibitor, 10 ?mol?L -1) + ADM(100 nmol?L -1) and PMA (a specific protein kinase C activator, 1 ?mol?L -1) on I Ca,L, respectively. RESULTS: ADM at the concentrations of 1-100 nmol?L -1 decreased I Ca,L in a dose-dependent manner (P

Objective To study the biological characteristics of platelet-derived growth factor A and human beta defensin 2 (hPDGF-A/hBD2) gene-modified rat bone marrow mesenchymal stem cells (BMSCs). Methods By using liposome transfection technique, recombinant adenovirus vector expressing hPDGF-A/hBD2 (Adv-hPDGF-A-IRES-hBD2) labeled with GFP was transfected into 293T cells for virus packaging and amplification. BMSCs were isolated, cultured and infected by adenovirus-containing supernatant. The exogenous gene-modified BMSCs were comprehensively studied on their biological features, in terms of morphology, cell growth curve, cell cycle, and adipogenic, osteogenic and myogenic differentiation ability. Results hPDGF-A-IRES-hBD2 gene-modified BMSCs did not show obvious changes in cell viability, proliferation, cell cycle distribution or cell differentiation. Conclusion BMSCs were not only good carriers for exogenous hPDGF-A and hBD2 genes but also seed cells for cell therapy even after hPDGF-A/hBD2 modification.

BACKGROUND:In order to solve the problems of poly-l-lactic acid (PLLA) stents, such as poor support, acidic metabolites, we researched a novel biogradable stent-PLLA/amorphous calcium plosphate (ACP).
OBJECTIVE: To discuss the safety and histocompatibility of the novel biogradable stent-PLLA/ACP stent implanted in the coronary artery in a porcine model.
METHODS:Sixteen novel biogradable stents were randomly implanted into the coronary arteries, left anterior descending branch, left circumflex artery or right coronary artery of sixteen healthy Tibet miniature pigs. The blood routine and blood biochemistry were measured pre-operation and at 1 month after operation. The coronary blood vessels where the stent was implanted were examined by hematoxylin-eosin staining at 1 and 6 months after operation.
RESULTS AND CONCLUSION: Compared with pre-operation, the post-operation indicators of the blood routine and blood biochemistry were of no significant difference. Coronary angiography revealed coronary artery patency and no thrombosis, the vascular stent segments exhibited clear boundaries with the surrounding tissue, with no tissue adhesion, necrosis, and adherence abnormalities. The results of hematoxylin-eosin staining showed that there was no significant difference in vascular injury integral between 1 month after operation and 6 months after operation. However, 6 months after operation, the scores of the inflammation were lower (P < 0.05), and the scores of the endothelialization were increased (P < 0.05). There was no myocardial infarction and inflammatory cellinfiltration around the stent. These results suggest that the novel biodegradable stent has good safety and histocompatibility.

Objective: To apply high-throughput whole genome sequencing (WGS) and short tandem repeat (STR) typing to detect aneuploidies, heteroploidies and copy number variations(CNVs) in spontaneous abortic tissues. Methods: Chorionic villus samples from 145 patients with spontaneous abortion were subjected to detection of aneuploidies, heteroploidies and copy number variations by WGS and STR typing. Results: All testing was successful and the rate of chromosomal abnormalities among the patients was 22.07%. Among these, there were 11 trisomies, 3 monosomies, 2 triploidies, 5 autosomal mosaicisms, 4 sex chromosomal mosaicisms, 7 structural abnormalities (including 1 mosaicism). In 89 cases, there were 130 CNVs of uncertain significance, 47 likely benign CNVs, and 2 loss of one copy of pathogenic AR gene. One sample contained 6 fragment duplications and deletions. Only 24 samples had no abnormal finding. Conclusion The most important reason for spontaneous abortions is embryonic chromosomal abnormality. Combined STR typing and WGS is both comprehensive and fast, and may become a major means for the detection of chorionic villi tissue from spontaneous abortions.

OBJECTIVE: To explore the genetic basis for a child with developmental delay and mental retardation. METHODS: Chromosomal karyotype of the child was analyzed by G-, C- and N-banding techniques. Her genome DNA was analyzed with single nucleotide polymorphisms array (SNP array). The result was validated by fluorescence quantitative polymerase chain reaction (PCR). RESULTS: The karyotype of the child was ascertained as 46,XX,r(22)(p12q13). SNP array has revealed a deletion of approximately 1.4 Mb at 22q13.33 (49 802 963-51 197 766). The deletion has encompassed the SHANK3, a crucial gene for the development of nervous system. Fluorescence quantitative PCR has confirmed the deletion of exons 7, 19 and 22 of the SHANK3 gene. CONCLUSION The phenotype of the patient may be attributed to the microdeletion at 22q13.33. Cytogenetic methods combined with SNP array and fluorescence quantitative PCR can identify aberrant chromosomes and provide accurate information for the clinical diagnosis and genetic counseling.

Objective To systematically evaluate the application effect of the quality control circle(QCC)in the complications after PICC catheterization.Methods The randomized controlled trials(RCTs)on the application effect of QCC after PICC catheter-ization were retrieved from the Cochane library,PubMed,EMBASE,Chinese Biomedical literature Database(CBM),China Academic Journal Full-text Database(CNKI),VIP and Wangfang Database by computer.The literatures were selected according to the inclu-sion and exclusion criteria.The two valuators independently retrieved and screened the literatures,extracted the data,evaluated the methodological quality of included literatures and conducted the cross check.Then the meta analysis was performed by using Rev-Man 5.3 software.Results A total of 7 914 articles were retrieved,finally 14 RCTs were included,involving 2 728 patients.The oc-currence rates of phlebitis,catheter obstruction,unplanned extubation and catheter-related blood flow infection in the intervention group were lower than those in the control group,the difference was statistically significant(OR=0.28,95% CI:0.16-0.48,P<0.01;OR=0.27,95% CI:0.17-0.42,P<0.01,OR=0.27,95% CI:0.18 -0.39,P<0.01;OR=0.25,95% CI:0.13 -0.49,P<0.01).Conclusion Using QCC is conducive to reduce the complication incidence rate after PICC catheterization.

OBJECTIVE: To explore the genetic cause for a child featuring growth and mental retardation. METHODS: Following conventional karyotyping analysis of the trio family, next generation sequencing (NGS) was carried out to explore the origin of the supernumerary marker chromosome. Fluorescence in situ hybridization (FISH) was used to confirm the result. RESULTS: The karyotypes of both parents were normal, while the proband was found to be 47,XX,+mar. NGS showed that the supernumerary marker has originated from chromosome 9p13.1p24.3 with a size of 39.77 Mb. FISH has confirmed the above finding. CONCLUSION The 9p13.1-p24.3 trisomy probably underlies the abnormal phenotypes of the child. Cytogenetic analysis combined with NGS and FISH can provide accurate diagnosis for such disorders.
Child ; Chromosomes, Human, Pair 9 ; genetics ; Cytogenetic Analysis ; High-Throughput Nucleotide Sequencing ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Trisomy

OBJECTIVETo assess the value of next generation sequencing (NGS) for the analysis of spontaneous abortion samples.

METHODSThe NGS analysis was carried out on 85 chorionic villi samples (taken between 42 days to 12 weeks of gestation) for which conventional cell culture has failed or chromosomal karyotyping has yielded normal or uncertain result.

RESULTSAmong 68 samples with a normal karyotype, the NGS analysis has identified 2 copy number variations (CNVs) and 2 chimeras. For 16 cases with failed cell culture, the NGS has identified 4 chromosomal abnormalities including 1 copy number variation and 3 numerical chromosomal aberrations. For 1 remaining case with uncertain karyotyping result, the NGS analysis has verified it as 46,XX,del(4) (p15.1p16.3).seq[GRCh37/hg19] (57 549 - 32 371 364)×1.

CONCLUSIONThe NGS analysis is capable of identifying novel CNVs in samples for which conventional cell culture may fail or karyotyping analysis may yield a normal result.

Abortion, Spontaneous ; genetics ; Adolescent ; Adult ; Cells, Cultured ; DNA Copy Number Variations ; Female ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Karyotyping ; Middle Aged ; Pregnancy ; Young Adult

OBJECTIVETo analyze the genetic cause for a child with growth retardation and mental retardation and discuss the application of array-based comparative genomic hybridization (aCGH) in its molecular genetic diagnosis.

METHODSConventional karyotyping of peripheral blood for the family was carried out. aCGH was performed to further ascertain the size and origin of the additional chromosome fragments.

RESULTSIn the trio family here, the karyotype of the father was normal, the karyotype of the mother was 46,XX, t(6;9)(q26;q21)and the proband child's was 47,XX,+der(9)?t(6;9)(q26;q21). aCGH showed that the extra chromosomal fragments originated from chromosome 9p24.3-q21.13 and the size was 78.26 Mb, and the repeat region included the 9p trisomy's clinical area. At the same time, it was confirmed that 6q26-q27 was trisomic and the fragment that related to development delay was 6.6 Mb. We determined that the proband's karyotype was 47,XX,+der(9)t(6;9)(q26;q21.13)mat finally.

CONCLUSIONThe patient's abnormal chromosome has originated from her mother with balance translocation. The duplications of 9p24.3-q21.13 and 6q26-q27 may lead to growth retardation and mental retardation. Accompanied with the cytogenetic methods, aCGH can accurately identify the origin and size of the abnormal chromosomes, contributing to the genetic analysis.

Child, Preschool ; Chromosome Disorders ; genetics ; Chromosomes, Human, Pair 6 ; genetics ; Chromosomes, Human, Pair 9 ; genetics ; Comparative Genomic Hybridization ; methods ; Female ; Humans ; Trisomy ; genetics

OBJECTIVE: To explore the genetic basis for a patient featuring developmental delay. METHODS: The patient and her parents were subjected to G- and C-banded chromosomal karyotyping analysis. The proband was also analyzed by single nucleotide polymorphism microarray (SNP-array). The result was verified by using fluorescence quantitative PCR (qPCR). RESULTS: The proband's karyotype was ascertained as 46,XX, r(15)(p11.2q26.3)[92]/45,XX,-15[9]/46,XX, dic r(15)(p11.2q26.3;p11.2q26.3)[4]. SNP-array revealed that she has carried a de novo deletion at 15q26.3 (98 957 555-102 429 040) spanning approximately 3.4 Mb, which encompassed the IGF1R gene. qPCR has confirmed haploinsufficiency of exons 3, 10 and 20 of the IGF1R gene. Both of her parents had a normal karyotype. CONCLUSION The abnormal phenotype of the proband may be attributed to the microdeletion at 15q26.3, in particular haploinsuffiency of the IGF1R gene and instability of the ring chromosome. Cytogenetic method combined with SNP-array and qPCR can efficiently delineate chromosomal aberrations and provide accurate information for clinical diagnosis and genetic counseling.
Chromosome Deletion ; Cytogenetic Analysis ; Female ; Genetic Counseling ; Humans ; Karyotyping ; Phenotype ; Ring Chromosomes