Alzheimer's disease(AD)is a multifactorial degenerative disorder of the central nervous system that mainly manifests as cognitive dysfunction and loss of speech.In recent years,the zebrafish has attracted extensive attention because of its high homology with humans in terms of brain structure and function,nerve conduction,and pathogenic genes of AD.This article reviews the advantages of the zebrafish as an animal model of AD,covering topics in the pathogenesis of AD and the evaluation and screening of drugs for treatment of AD.The overall goal is to provide new insights into the pathogenesis of AD and development of novel drugs.

Objective:Through the investigation of the pathogenicity of COL4A4 heterozygous splicing mutations and the genotype-phenotype correlation in autosomal dominant Alport syndrome (ADAS), to better understand the impact of COL4A4 heterozygous splicing mutations on ADAS. Methods:The study was a case series analysis. Patients from 5 ADAS families with COL4A4 heterozygous splicing mutations detected by whole exome sequencing were recruited by three hospitals. In vivo transcriptional analysis and/or in vitro minigene splicing assay were conducted to determine the splicing patterns and assess the pathogenicity of COL4A4 heterozygous splicing mutations. Results:In the five ADAS pedigrees carrying COL4A4 heterozygous splicing mutations, four novel ADAS splicing patterns were described. In pedigree 1-4, most patients presented with continuous hematuria or/and microalbuminuria. Otherwise,the proband in pedigree 4 presented with macroalbuminuria and the proband in pedigree 1 had progressed to chronic kidney disease stage 2 at the age of 70 years old. In pedigree 5, all patients developed end-stage renal disease between 28 and 41 years old. c.735+3A>G detected in pedigree 1 and pedigree 2 and c.694-1G>C detected in pedigree 3 both led to exon 12 skipping in COL4A4, resulting in 42 nucleotides in-frame deletion (c.694_735del). c.2056+3A>G detected in pedigree 4 led to COL4A4 exon 26 skipping, which caused in-frame deletion of 69 nucleotides (c.1988_2056del). c.2716+5G>T detected in pedigree 5 led to a 360 nucleotides large in-frame deletion, including 100 bp sequence at the 3'end of exon 29,the whole sequence of exon 30 and 89 bp sequence at the 5'end of exon 31 (c.2446_2805del). Conclusions:Renal prognosis differs significantly for patients with small in-frame deletions versus large in-frame deletion splicing abnormalities. Determination of the pathogenicity and the splicing patterns of COL4A4 heterozygous splicing mutations using in vivo and in vitro transcriptional analysis may provide renal prognostic information.

Aromatase deficiency (AD) is a rare autosomal recessive genetic disease caused by loss-of-function mutations in aromatase gene (CYP19A1), leading to congenital estrogen deficiency syndrome. Both mothers of AD patients during pregnancy and female AD fetus show virilization, while male patients are usually diagnosed in adulthood due to continued height increase and metabolic abnormalities. In 2019, a patient with AD was admitted in the Second Xiangya Hospital. The patient was a 37-year-old adult male who continued to grow linearly after adulthood. His estradiol was below the measurable line, the follicle-stimulating hormone (FSH) increased, bone age delayed, epiphysis unfused, and the bone mass reduced. CYP19A1 gene detection showed that c.1093C>T, p.R365W was homozygous mutation. This disease is rare in clinic. Clinicians need to raise awareness of the disease for early diagnosis and treatment to improve the long-term prognosis of patients.
46, XX Disorders of Sex Development/genetics* ; Adult ; Aromatase/metabolism* ; Female ; Gynecomastia/genetics* ; Humans ; Infertility, Male ; Male ; Metabolism, Inborn Errors ; Mutation ; Pregnancy

The aim of the study was to develop a simple, rapid and accurate LC-MS/MS method for the determination of digoxin.Digoxin-d3 was taken as the internal standard (IS), and sample preparation was achieved by liquid-liquid extraction.Chromatographic separation was performed on a Kinetex C18 column (2.1 mm × 50 mm, 2.6 μm; Phenomenex) using an isocratic elution with merely 2 min for each sample.The mobile phase consisted of water and acetonitrile solutions, both containing 1 mmol/L ammonium acetate and 1 mmol/L formic acid (55∶45).The detection was conducted on a TripleQuadTM 4500MD mass spectrometer coupled with electrospray ionization interface under positive-ion multiple reaction monitoring mode.The transitions were m/z 798.5 → 651.3 and m/z 801.6 → 654.4 for digoxin and digoxin-d3, respectively.Results showed that the method was linear over the range of 0.100-20.0 ng/mL.The selectivity, accuracy and precision, recovery and stability of the method were all within the acceptable limits with no matrix effect.This method was successfully applied to a girl treated with digoxin with substantial improvement of therapeutic effect and elimination of toxic reaction, so it can provide valuable fuidance and reference for individualized medication in clinical practice.

Objective To explore the clinical efficacy and safety of recombinant human brain natriuretic peptide (rhBNP) in the treatment of acute heart failure(AHF) by observing the changes of hemodynamic parameters,cardiac function and inflammatory factors before and after treatment in the patients with AHF.Methods A total of 96 patients with AHF in our hospital from June 2012 to December 2015 were enrolled in this study and divided into the control group and observation group(n-48).The two groups were given the routine anti-heart failure treatment combined with sodium nitroprusside or rhBNP by intravenous dripping for 24 h.Fifteen cases were selected from each group for monitoring the hemodynamic change.The clinical effect was observed.The changes of heart rare,blood pressure,urine volume,cardiac function,plasma of NT-proBNP,IL-6 and hs-CRP before and after medication were observed.The occurrence of adverse reactions was also observed.Results The total effective rate of the observation group was significantly higher than that in the control group (P＜0.05).The PAP and PCWP at various time points after treatment in the observation group were significantly lower than those before treatment (P＜0.05).PAP and PCWP at various time points had statistical difference between two groups (P＜0.05).The levels of heart rate,systolic blood pressure,NT-proBNP,IL-6 and hsCRP levels after medication were significantly decreased in both groups,and the urine volume and LVEF were significantly increased (P＜0.05),and the difference between the two groups was statistically significant(P＜0.05).There was no statistically significant difference in the occurrence of adverse reactions between the two groups (P＞0.05).Conclusion The short-term efficacy of rhBNP in treating AHF is better than sodium nitroprusside,which can improve hemodynamics and cardiac function,reduces the level of inflammatory factors.

Objective To apply color Doppler flow imaging (CDFI) and transcranial Doppler (TCD) to the follow-up observation of the changes of cervical vessel,intracranial hemodynamics and cerebrovascular reserve capacity (CVR) of the patients after carotid artery stent implantation.Methods Totally 96 patients with carotid artery stent implantation underwent CDFI and TCD examinations,and the changes of hemodynamics were compared before and 1 month,6 months,1 a and 2 a after implantation.Results The values of peak systolic velocity (PSV) and resistance index (RI) at the areas of carotid stenosis were lower significantly than those before implantation,while the values of PSV,pulsatility index (PI) and CVR of the middle cerebral artery were obviously higher than those before implantation (P＜0.05).There were no significant differences between the cervical and intracranial hemodynamics indexes 1 month,6 months,1 a and 2 a after treatment.Two-year follow-up found 4 cases of restenoses after implantation,and the rate for restenosis was 4.2％.Conclusion Carotid artery stent implantation improves significantly cervical and intracranial blood supply as well as CVR of the carotid stenosis patient,and CDFI combined with TCD can be used for the accurate evaluation of the efficacy and postoperative follow-up of carotid artery stent implantation.

Objective To understand the prevalence of birth defects, related diseases and mental status of women during pregnancy in Shaanxi province and to analyse the major risk factors on birth defects and congenital heart disease. Possible association between maternal diseases or mental status and the risk of birth defects, was also explored. Methods A cross-sectional design was used in this study and stratified multistage random sampling method was used. The whole survey was from Jury 2013 to November 2013. Logistic regression method was used to analyze the association between maternal diseases, mental status during pregnancy and birth defects. Results The overall prevalence of birth defects was 195.04 per 10000 in Shaanxi. Among the 29121 mothers participating in this study, 51.1% developed illness and 6.8%"changed their mental status during pregnancy. After adjusting all the confounding factors, results showed that, histories of cold", fever, and intrahepatic cholestasis were (OR=1.33, 95%CI:1.10-1.61, OR=1.54, 95%CI:1.09-2.16, and OR=32.77, 95%CI:4.08-263.04) respectively, during pregnancy that related to birth defects. Self-reported unstable mental status (OR=1.60, 95%CI: 1.19-2.15) and family friction (OR=2.07, 95%CI: 1.12-3.79) were both related to the birth rates. Histories of cold and fever (OR=1.59, 95%CI:1.28-1.98;OR=1.43, 95%CI:1.48-4.00), during early pregnancy, unstable mental status during mid-pregnant period (OR=1.52, 95%CI:1.05-2.19), unstable mental status during late-pregnant period (OR=1.63, 95%CI:1.05-2.19) and family friction during late-pregnant period (OR=2.89, 95%CI:1.16-7.20) were found to be related to birth defects. Compared with those without history of cold, those with the history of cold during first (OR=1.24, 95%CI: 1.02-1.52) and second stages (OR=2.06, 95%CI: 1.30-3.26) of pregnancy were more likely to bear fetus with birth defects. Compared with those without these histories, those with histories of fever (OR=1.49, 95%CI:1.04-2.13), emotional problem (OR=1.71, 95%CI:1.19-2.45) and related diseases (OR=2.67, 95%CI: 1.32-5.39) during the first period of pregnancy were more likely to bear fetus with birth defects. Conclusion The incidence of birth defects in Shaanxi was high. Histories of cold, fever, unstable mental status and family friction during pregnancy, seemed to have increased the risks of bearing child with birth defects.

OBJECTIVE: To establish the cell model using human leukemia cell line HL-60 for exposure of coke oven emissions( COE) in vitro and to explore the mechanism of COE-induced acute toxicity in HL-60 cells. METHODS: HL-60 cells were collected in their logarithmic growth phase and cultured in medium that had final concentrations of COE in 2. 5,5. 0,10. 0 and 20. 0 mg / L for 24 hours. Cell survival rate was examined by CCK-8 assay. The cytotoxicity was evaluated using lactate dehydrogenase release assay. Reactive oxygen species( ROS) production was determined by the 2',7'-dichlorofluorescein diacetate and nitroblue tetrazolium method. The activation of nuclear factor-κB( NF-κB) pathway was evaluated by western blot. RESULTS: With the increasing exposure concentrations of COE,the cytotoxicity of HL-60 cells increased( P < 0. 01),the cell survival rate decreased( P < 0. 01),intracellular ROS decreased( P < 0. 01),whereas extracellular ROS increased( P < 0. 01). These changes had a dose-effect relationship. The levels of phospho-nuclear factor-kappa B p65 and phospho-inhibitor of kappa Bα were higher in all the COE-treated cells compared with untreated cells( P < 0. 05),with no dose-effect relationship. CONCLUSION: COE could cause acute toxicity in HL-60 cells in a doseeffect relationship. The mechanism may be related to the COE-induced in-balanced ROS release and removal,leading to the activation of NF-κB pathway. HL-60 cells can be used as a common cell line for COE hematotoxicity analysis.

Anemia, Sideroblastic ; diagnosis ; genetics ; Asian Continental Ancestry Group ; Genetic Diseases, X-Linked ; diagnosis ; genetics ; Humans ; Pedigree

Objective To compare the effect of different positive end-expiratory pressures (PEEPs) on the efficacy of volume therapy guided by global end-diastolic volume index (GEDVI) and central venous pressure (CVP) in patients with septic shock.Methods Twenty-five patients with septic shock complicated with respiratory failure,of both sexes,aged 18-64 yr,were enrolled in the study.Their APACHE [[scores were 13-31.The patients were endotracheally intubated and underwent volume-controlled ventilation,PEEP was 5-15 cmH2O,and pulse oxygen saturation was maintained ＞ 90 ％.The patients were divided into low PEEP (5-10 cmH2 O) group and high PEEP (11-15 cmH2 O) group depending on the different PEEP levels.6 ％ hydroxyethyl starch (200/0.5)6 ml/kg was infused over 30 min for volume therapy.Right internal jugular vein or subclavian vein was cannulated for CVP monitoring,and GEDVI was continuously monitored by pulse indicator continuous output monitoring (PiCCO) technology.CVP and GEDVI were recorded before and after volume therapy and the changing rate was calculated.Results Compared with CVP and GEDVI before volume therapy,CVP and GEDVI were significantly increased after volume therapy in low PEEP group (P ＜ 0.05),and GEDVI was increased after volume therapy (P ＜ 0.05) and no significant change was found in CVP after volume therapy in high PEEP group (P ＞ 0.05).Compared with low PEEP group,the changing rate of CVP was significantly decreased (P ＜ 0.05),and no significant change was found in the changing rate of GEDVI in high PEEP group (P ＞ 0.05).Conclusion High PEEP can decrease the efficacy of volume therapy guided by CVP,while exerts no effect on the efficacy of volume therapy guided by GEDVI in patients with septic shock.