OBJECTIVE: To evaluate and compare whole exome sequencing (WES) and targeted panel sequencing in the clinical molecular diagnosis of the Chinese families affected with inherited retinal dystrophies (IRDs). METHODS: The clinical information of 182 probands affected with IRDs was collected, including their family history and the ophthalmic examination results. Blood samples of all probands and their relatives were collected and genomic DNA was extracted by standard protocols. The first 91 cases were subjected to the WES and the other 91 cases were subjected to a specific hereditary eye disease enrichment panel (HEDEP) designed by us. All likely pathogenic and pathogenic variants in the candidate genes were determined by Sanger sequencing and co-segregation analyses were performed in available family members. Copy number variations (CNVs) detected by HEDEP were further validated by multiplex ligation-dependent probe amplification (MLPA). As PRGR ORF15 was difficult to capture by next generation sequencing (NGS), all the samples were subjected to Sanger sequencing for this region. All sequence changes identified by NGS were classified according to the American College of Medical Gene-tics and Genomics and the Association for Molecular Pathology (ACMG/AMP) variant interpretation guidelines. In this study, only variants identified as pathogenic or likely pathogenic were included, while those variants of uncertain significance, likely benign or benign were not included. RESULTS: In 91 cases with WES, pathogenic or likely pathogenic variants were determined in 30 cases, obtaining a detection rate of 33.00% (30/91); While in 91 cases with HEDEP sequencing, pathogenic or likely pathogenic variants were determined in 51 cases, achieving the diagnostic rate of 56.04% (51/91), and totally, the diagnostic rate was 44.51%. HEDEP had better sequencing coverage and read depth than WES, therefore HEDEP had higher detection rate. In addition, HEDEP could detect CNVs. In this study, we detected disease-causing variants in 29 distinct IRD-associated genes, USH2A, ABCA4 and RPGR were the three most common disease-causing genes, and the frequency of these genes in Chinese IRDs population was 11.54% (21/182), 6.59% (12/182) and 3.85% (7/182), respectively. We found 43 novel variants and 6 cases carried variants in RPGR ORF15. CONCLUSION NGS in conjunction with Sanger sequencing offers a reliable and effective approach for the genetic diagnosis of IRDs, and after evaluating the pros and cons of the two sequencing methods, we conclude that HEDEP should be used as a first-tier test for IRDs patients, WES can be used as a supplementary molecular diagnostic method due to its merit of detecting novel IRD-associated genes if HEDEP or other methods could not detect disease-causing va-riants in reported genes. In addition, our results enriched the mutational spectra of IRDs genes, and our methods paves the way of genetic counselling, family planning and up-coming gene-based therapies for these families.
DNA Copy Number Variations ; Humans ; Mutation ; Pedigree ; Retinal Dystrophies/genetics* ; Whole Exome Sequencing

Cytokines ; Hashimoto Disease ; diagnosis ; prevention & control ; Humans

Objective To investigate whether Shenfu(SF) injection has protective effect on circulation function in infants undergoing cardio-pulmonary bypass.Methods Thirty-six patients within 3 years old with congenital heart disease were randomly divided into SF injection group(n=18) and control group(n=18).The SF group were treated with 1 mL/kg SF intravenous injection after inducing of anesthesia,the control group were infused with 1 mL/kg saline intravenously.The changes of hemodynamics during surgery,time needed for sinus rhythm emerging on electrocardiogram(ECG) after arteriae aotra patency dose of vaso-active drugs intra-and postoperatively and recovery time during postoperative period were observed.Results In SF group,mean artery pressure(MAP) was higher and heart rate(HR),central venous pressure(CVP) lower than control group(P

Objective To explore the relationship between predictable ocular motor control and cognitive function in Parkinson's disease(PD). Methods Videonystagmography was used to examine 24 patients with idiopathic PD(PD group) and 24 healthy controls(control group) on predictable ocular motor control.The accuracies of saccade were compared between two groups.The correlation among accuracy for predictive saccade(latency

There has been a series of teaching innovation carried out to improve the clinical teaching of seven-year program students in the department of gynecology and obstetirics of Xinqiao hospital.By using case as a guide and problem as foundation,with multimedia and other advanced teaching plan,we paid more attention to the Chinese-English teaching to improve mixed diathesis of these medical students and find out a better teaching model.

Objective:To develop a liquid chromatography/tandem mass spectrometry employing precolumn derivatization method for determination of peramivir in rat plasma.Methods: The sample preparation consisted of a protein precipitation extraction followed by derivatization with hydrochloric acid (10 mol/L) methanol (10∶90, v/v) and determined by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The positive ion SRM mode was performed and the precursor-to-the-product ion transitions of m/z 343→284 and 299→152 were used to measure the derivative of peramivir and Ro 64-0802 (I.S.). The chromatographic separation was achieved using a Zorbax RX-C8 (2.1 mm × 150 mm × 5 μm) analytical column with an isocratic mobile phase composed of acetonitrile-water-formic acid (30∶70∶0.1, v/v/v, 0.2 ml/min). Results:The method was linear over a concentration range of 10 to 10 000 ng/ml. The LLOQ was 10 ng/ml. The average inter-day/intra-day precision values ranged from 5.0% to 7.1%, respectively, while the accuracy values ranged from 89.9% to 106.1%.Conclusion: In this method, retention time is greatly improved, the matrix effects are decreased by chemical derivatization. This method has been successfully applied to the preclinical and clinical research of peramivir.

Asthma ; physiopathology ; Child ; Female ; Humans ; Male ; Maximal Expiratory Flow-Volume Curves ; Oscillometry ; methods ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; Respiratory Function Tests

Objective:To compare psychosocial factors and immune function between patients with breast cancer and benign tumor before operation.Methods:The patients of breast tumor(56 case)were measured by clinical psychological rating scale(EPQ、HAMD、HAMA、LES、SSRS、DSQ)before operation,and their serum levels of IgA, CD_4~+,CD_(19)~+ and the activity of NK cell were measured.The patients of breast tumor were divided into breast cancer group(36 case)and benign tumor group(22 case)bases on results of pathology.Results:In breast cancer group, there were significantly higher scores of neurotic,total and negative life events,immature defense styles,HAMA and HAMD than those in benign tumor group;while NK cell activity,levels of CD_4~+ and CD_(19)~+ and the score of mature defense,social supports,subjective supports and utility degree of supports were marked lower.There were significantly correlation between immune function(NK cell activity,content of CD_4~+,CD_(19)~+,IgA)and psychosocial factors(social support,P score of EPQ,HAMD and HAMA total score,quantity of negative life events etc). Conclusion:Cancer patients had more server psychosocial problems and lower immune function.The immune function of patient with breast cancer before operation is associated with the various psychosocial factors.

China ; Gastroesophageal Reflux ; therapy ; Humans ; Integrative Medicine ; Medicine, Chinese Traditional ; methods

Adult ; Chromosome Aberrations ; Chromosome Disorders ; epidemiology ; Gold ; Humans ; Lymphocytes ; Male ; Middle Aged ; Mining ; Occupational Exposure ; Young Adult