Objective To study the changes and clinical significance of heart rate turbulence(HRT)in patients with diabetic cardiomyopathy.Methods 24h Holter ECG recordings were examined in 33 patients with diabetic cardiomyopathy(DC)and 30 patients in the control group.Turbulence onset(TO)and turbulence slope (TS)were calculated from Holter recordings.The mean value of TO,TS between the two groups were compared,the relativity among TO,TS,SDNN and LEVDD were analyzed as well.Results TO was significantly higher in the DC group compared with the control group[(-0.38 ± 0.12)％ vs(-1.88 ± 0.13)％,t =7.38,P =0.01)].TS in the DC group was significantly lower than that in the control group[(2.02 ± 0.82)ms/R-R vs (9.72 ± 1.18)ms/R-R,t =18.69,P =0.02)].TO was significantly related to SDNN(r =-0.35,P =0.02)and LEVDD(r =0.68,P =0.01),TS was also significantly related to SDNN(r =0.45,P =0.01)and LEVDD (r =-0.39,P =0.03).Conclusion The manifest of heart rate turbulence is significantly impaired in patients with diabetic cardiomyopathy.

Objective To explore sinus heart rate turbulence (HRT) in patients with different subtypes of essential hypertension (EH), and analyze the relationship between HRT and autonomic nervous system function damage in these patients. Methods The study consisted of 107 patients with EH (EH group) and 46 controls (control group). Based on 24 hours dynamic electrocardiogram, all patients were divided into dipper,non-dipper,and anti-dipper blood pressure group. The indexes about HRT and heart rate variability (HRV) among these groups were calculated and compared,and the relationship between turbulence onset (TO),turbulence slope (TS) and 24 hours mean systolic blood pressure,diastolic blood pressure was analyzed. Results There were significant differences in TO,TS,SDNN between EH group and control group(P ＜ 0.05 ). TO in non-dipper and anti-dipper blood pressure group was significantly higher than that in control group( P ＜ 0.05 ), and TS was lower than that in control group(P ＜ 0.05 ). There was no significant difference in TO,TS between dipper blood pressure group and control group (P ＞ 0.05). TO in non-dipper and anti-dipper blood pressure group was significantly higher than that in dipper blood pressure group, but TS was lower than that in dipper blood pressure group (P ＜0.05). Correlation analysis showed that TO had positive relationship with average heart rate and age (rs = 0.265, P = 0.004;rs = 0.217, P = 0.018 ), but had negative correlation with SDNN and left ventricular ejection fraction (LVEF) (rx = -0.287,P = 0.002;rx =-0. 179, P = 0.049). Whereas, TS had negative correlation with average heart rate and age (r = -0.335, P =0.015 ;r = -0.238,P= 0.009), but had positive relationship with SDNN and LVEF(r = 0.540,P = 0.001 ;r =0.432,P = 0.001 ). Conclusions HRT of EH patients becomes significantly low. It suggests that the autonomic nerve function in EH patients be injured seriously.

Objective To construct a lentiviral vector carrying angiopoietin-1 (Ang-1) and DsRed gene, and to package a virus particles. Methods The Ang-1 lentiviral vector with DsRed (PLV.Ex3d.P/puro-CMV>Ang-1>IRES/DsRed-Express2) was constructed by Gateway technology, and identiifed by PCR and gene sequencing. The lentiviral vector was mixed with helper vector pLV/helper-SL3, pLV/helper-SL4 and pLV/helper-SL5 by Lipofectamine 2000 to prepare DNA-Lipofectamine?2000 complexes. The complexes were then added to transfect 293FT cells and package virus. The virus titers and infection ef-ifciency were determined by lfuorescence expression. Results Ang-1 lentiviral vector PLV.Ex3d.P/puro-CMV>Ang-1>IRES/DsRed-Express2 was constructed successfully as identified by PCR and gene sequencing. Lentivirus with high-efficiency infection was produced by transfection to 293FT cells and the virus titer was 5×108 TU/ml. Conclusions The recombinant len-tiviral vector for Ang-1 was successfully constructed by Gateway technology and the lentivirus with high-efifciency infection packaging can be used for further experiment of Ang-1 gene.

OBJECTIVETo explore the candidate disease causing gene for a case with floppy infant syndrome (FIS).

METHODSSingle nucleotide polymorphism array (SNP array) was used for analyzing the whole genome copy number mutations in the proband. Multiple PCR combined with denaturing high performance liquid chromatography (DHPLC) was employed to verify the suspected mutations in the proband and his family members.

RESULTSA large duplication arr [hg19] Xq13.1: 67 987 646-73 805 828, which spans approximately 5.818182 Mb and encompasses 66 known genes, was identified in the proband. The multiple PCR-DHPLC assay confirmed duplication of HDAC8, PHKA1, TAF1, DLG3, KIF4A, IGBP1, PJA1 and SLC16A2 genes in the proband. His mother and grandmother both had duplication of the above genes in one X chromosome, but his aunt had not.

CONCLUSIONThe large Xq13.1 duplication identified by the SNP array probably underlies the FIS in this family. For its high-throughput, high resolution and capacity of automation, SNP array has provided a first line method for the genetic testing for infants featuring developmental delay with unknown reason, mental retardation, autism, multiple malformation and FIS.

Objective: To investigate the association of iron with stress hormones and insulin resistance in patients with gestational diabetes mellitus (GDM). Methods: Seventy-five pregnant women diagnosed as GDM during 24-28 weeks were collected from January to November 2015 in Yantai Yuhuangding Hospital, and 75 normal pregnant women were used as control group. Blood glucose, insulin, stress hormones and iron metabolism related indexes were detected. Homeostasis model assessment for insulin resistance (HOMA-IR) was used to evaluate insulin resistance, and the correlation of iron metabolism with stress hormones and insulin resistance was analyzed. Results: Compared with control group, norepinephrine (NE) and epinephrine (E) levels were higher in GDM group (both P<0.05), but there was no significant difference in cortisol level between two groups (P>0.05). Serum ferritin (SF), serum iron, and transferrin saturation levels were higher in GDM group than those in control group(all P<0.01). Multivariate logistic analysis showed that cortisol, E, and NE levels were positively correlated with SF level in GDM group (all P<0.05). Cortisol and E levels were positively correlated with transferrin saturation (both P<0.05). SF and transferrin saturation were positively correlated with HOMA-IR in two groups (P<0.05 or P<0.01). Conclusion Iron overload may involve in dysfunction of stress adaptation and insulin resistance in patients with GDM. (Chin J Endocrinol Metab, 2018, 34: 563-566)

OBJECTIVETo develop a simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of coxsackievirus A6 (CV-A6) based on the colour chang of hydroxy naphthol blue (HNB).

METHODSThe method employed a set of six primers that recognized sequences of VP1 gene for amplification of nucleic acid under isothermal conditions at 63 °C for 50 min. The products were detected through visual inspection of color change by the pre-addition of HNB dye. The specificity was validated by detecting a collection of different human enteroviruses. The sensitivity of this assay was evaluated by serial dilutions of RNA molecules from in vitro transcription of CV-A6 VP1 gene, and compared with real-time RT-PCR (rRT-PCR) in parallel. This assay was evaluated with 92 clinical specimens from patients with hand-foot-mouth disease.

RESULTSA positive color (sky blue) was only observed in the preparation of CV-A6, whereas none of the other 23 kinds of human enteroviruses showed a color change. The HNB based RT-LAMP showed a sensitivity of 100 copies/reaction, which was at the same level as that of the rRT-PCR. The result of RT-LAMP in analysis of 92 clinical specimens was consistent with that of the rRT-PCR. The kappa correlation between the two methods was 1 and both of the sensitivity and specificity of the RT-LAMP assay were 100%.

CONCLUSIONThe established RT-LAMP assay had good specificity and sensitivity and thus demonstrated to be a promising screening tool for CV-A6 infection. It also has the potential to be used in resource-limited clinical sites and field study.

Colorimetry ; Coloring Agents ; chemistry ; DNA Primers ; Enterovirus ; isolation & purification ; Hand, Foot and Mouth Disease ; virology ; Humans ; Indicators and Reagents ; chemistry ; Naphthalenesulfonates ; chemistry ; Nucleic Acid Amplification Techniques ; Real-Time Polymerase Chain Reaction ; Reverse Transcription ; Sensitivity and Specificity

Objective To analyze the clinical manifestation and genetic testing in a patient with Adams-Oliver syndrome (AOS) and summarize clinical and genetic characteristics of the dedicator of cytokinesis (DOCK) 6 gene related AOS through reviewing related references.Methods Information of the proband who was hospitalized in Affiliated Children Hospital of Capital Institute of Pediatrics in October 2016 and her family members as well as their DNA samples were collected.The gene sequencing was performed using next generation sequencing technology.Using "Adams-Oliver syndrome"and "DOCK6" as key words,the relevant articles were searched from the Pubmed,China National Knowledge Internet and Wanfang databases and reports of 19 cases were reviewed.Results The proband is an eight months old girl.She presented with severe developmental delay,terminal transverse limb defects and visual loss after birth,and then suffered from tonic seizures and myoclonic seizures at two months old.By physical examination she was found to have esotropia and visual loss.The distal phalanx and nail of the right second-fourth fingers were absent,while the phalangette of the left second-fourth fingers and bilateral distal phalanges of toes were short with small nails attachment.Thyroid function test showed hypothyroidism.The ocular fundus examination showed the residual vitreous artery in the left eye and the retinal pigment degeneration in the right eye.CT scan showed multiple bilateral periventricular calcification and cranial magnetic resonance imaging showed bilateral periventricular lesion.Two heterozygous mutations were identified in DOCK6 gene:one was a known pathogenic mutation (p.L1064Vfs*60),and the other was a novel splice site mutation (c.873+ 1G＞A).By analyzing this case and reported 19 cases,the common performances of DOCK6 gene related AOS included terminal transverse limb defects (20/20),aplasia cutis congenita (18/20),ocular abnormalities (13/20),seizures (12/20),mental retardation (12/20),microcephaly (10/20),cardiovascular malformations (10/20),intrauterine growth retardation (6/20).The mutation of the DOCK6 gene was found to be dominated by frameshift mutation and splice site mutation.Conclusions If either terminal transverse limb defects or aplasia cutis congenita was detected in a patient,AOS should be under consideration.In addition,autosomal recessive inheritance,nervous system and eyes involvement will further indicate DOCK6 gene related AOS.

新冠病毒感染疫情严重威胁着世界各国人民的生命健康。目前，对病毒感染的防治研究主要集中在抑制病毒与分子受体的结合上。AXL作为新发现的严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）受体，在协助病毒感染人体呼吸系统中发挥着重要作用，是未来临床干预的潜在靶点。本研究对已发表的单细胞测序数据进行整理和分析，发现AXL在年轻人肺细胞中的表达水平明显高于老年人。人胚肺二倍体成纤维细胞（2BS）是衰老研究的公认细胞株。本文采用2BS细胞构建复制性细胞衰老模型，发现年轻细胞中AXL的蛋白水平明显高于衰老细胞，据此推测年轻人感染的风险可能更高，需要注意防护。我们发现一种羊栖菜褐藻多糖硫酸酯组分（SFW-3）可显著下调年轻2BS细胞中AXL的表达水平，表明SFW-3具有一定的抗SARS-CoV-2感染的研究价值，同时表明2BS细胞株也可作为潜在的SARS-CoV-2体外感染模型。
Humans ; SARS-CoV-2 ; Sargassum/metabolism* ; Diploidy ; Sulfates/metabolism* ; COVID-19 ; Polysaccharides/pharmacology* ; Lung