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 identify a pathogenic strain JM-1 isolated from the pus of a patient stabbed by a sea shrimp and to analyze its antibiotic susceptibility and virulence genes, aiming to provide reference for screening clinically related infections caused by Cysteiniphilum litorale as a rare pathogen and improving prognosis. Methods:Biochemical phenotype identification, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), 16S rRNA gene sequencing, analysis of average nucleotide identity (ANI) and average amino acid identity (AAI) based on the whole genome and phylogenetic analysis of the 16S rRNA gene and the whole genome were performed to accurately determine the taxonomic status of the strain JM-1. E-test was used to detect antibiotic susceptibility, and the results were interpreted according to the interpretation standards of Francisella tularensis in CLSI M45-A3. The virulence factor database (VFDB) was used for genome-wide annotation and analysis of virulence genes. Results:After culturing the strain JM-1 on the Columbia blood plate for 3 d, some grey-white, medium-sized, smooth, round and convex hemolytic colonies were observed. Gram staining result showed lightly colored Gram-negative Coccobacillus. API NH identification results suggested that the isolate JM-1 was Moraxella catarrhalis (biochemical code: 3010), while there was no identification result in Vitek2 system NH card (biochemical code: 0211002121). The EXS3000 mass spectrometer self-built database identified the isolate JM-1 as Cysteiniphilum litorale. The phylogenetic analysis based on the 16S rRNA gene and the whole genome showed that the isolate JM-1 and Cysteiniphilum litorale DSM 101832 T clustered into the same branch, and the ANI and AAI values between the two strains were 95.07% and 95.65%, respectively. The biochemical phenotype identification indicated the isolate JM-1 producing β-lactamase and penicillinase. Antibiotic susceptibility test results showed the strain was resistant to penicillin and sensitive to gentamicin, streptomycin, doxycycline, tetracycline, ciprofloxacin, levofloxacin, and chloramphenicol. Genome annotation suggested the virulence genes of the isolate JM-1 were similar to those of Francisella, including Francisella pathogenicity island (FPI), type Ⅳ fimbriae, capsule and lipopolysaccharide. Conclusions:Cysteiniphilum litorale was a rare pathogen with virulence genes similar to those of Francisella, and its antibiotic susceptibility was also similar to that of Francisella. This study confirmed a case of clinical infection caused by Cysteiniphilum litorale. The self-built MALDI-TOF MS system could be used for its rapid identification.

BACKGROUND:Nanocomposite hydrogel has great research prospects and application potential in the treatment of osteoarthritis. OBJECTIVE:To review the research progress of nanocomposite hydrogel in osteoarthritis and cartilage repair. METHODS:Databases such as CNKI and PubMed were searched.The English key words were"nanocomposite hydrogel,nanogel,osteoarthritis,cartage,physical encapsulation,electrostatic interaction,covalent crosslinking",and the Chinese key words were"nanocomposite hydrogel,nanogel,osteoarthritis,cartage,physical encapsulation,physical encapsulation,electrostatic effect,covalent cross-linking".After an initial screening of all articles based on inclusion and exclusion criteria,71 articles with high correlation were retained for review. RESULTS AND CONCLUSION:In cell or animal experiments,nanocomposite hydrogel has the effect of improving osteoarthritis.Nanocomposite hydrogel can promote cartilage repair,improve the internal environment of osteoarthritis,and achieve the therapeutic purpose of osteoarthritis by improving the mechanical environment between joints,carrying targeted drugs,and promoting the chondrogenesis of seed cells.At present,the research of nanocomposite hydrogel in osteoarthritis disease still has a huge space to play.It is expected to open up a new way for the clinical treatment of osteoarthritis by continuing to deepen the research of material preparation and actively carrying out cell and animal experiments.

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