Objective:To report four male COL4A5 mutation mosaicism patients with X-linked Alport syndrome, and to provide evidence for diagnosis, genetic counseling, and reproduction in the respective families and improve our knowledge of mosaicism in Alport syndrome. Methods:Suspected male mosaic patients for COL4A5 who met the following criteria: clinical diagnosis of Alport syndrome, harbored COL4A5 mutations detected using next generation sequencing or Sanger sequencing, heterozygosity for the mutant and normal COL4A5 alleles in the DNA demonstrated by Sanger sequencing, registered in the on-line registry of hereditary kidney diseases, and admitted to Peking University First Hospital during the period of April 2018 to April 2019 were enrolled. Clinical data and karyotypes were retrospectively analyzed. Genetic DNA isolated from multiple tissues was analyzed for COL4A5 gene mutations by using PCR and Sanger sequencing. Related literatures published in PubMed, CNKI and Wanfang databases were reviewed. Results:Four COL4A5 somatic and germline mosaic male patients with Alport syndrome were included in the study. Patient 1 was characterized by hematuria and proteinuria. His karyotype of peripheral blood was normal. COL4A5 c.3455-1G>A mosaicism was detected in multiple tissues (peripheral blood, saliva and urine). Patient 2 presented with hematuria and microalbuminuria. His karyotype of peripheral blood was normal. COL4A5 c.4994+1G>A mosaicism was detected in multiple tissues (peripheral blood, saliva and skin fibroblasts). Patients 3 showed hematuria without proteinuria. COL4A5 c.3535G>A mosaicism was found in genomic DNA of peripheral blood and hair. Laboratory and physical examinations of patient 4 showed hematuria and normal renal function, without proteinuria, megasoma or small testes. COL4A5 c.3106G>A mosaicism was detected in genomic DNA of skin fibroblasts. Although without karyotype analysis due to unavailable specimens, 47,XXY or 46,XY/47,XXY mosaicism was not considered according to the reproductive history and lack of clinical manifestations of megasoma and small testes in patients 3 and 4. Renal disease in 8 published male cases with mosaicism for COL4A5 was affected by mutant allelic fractions and genotype. Conclusions:Compared with hemizygous males with X-linked Alport syndrome, the renal phenotype of mosaic males was milder, and associated with mutant allelic fractions and mutation type.

Objective:To explore the application value of whole genome and high resolution chromosome microarray analysis (CMA) in genetically etiological diagnosis of infants and young children with congenital heart disease (CHD).Methods:The clinical data of 130 infants and young children with CHD who were hospitalized and received CMA test at the Department of Pediatrics, Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from January 2016 to December 2018 were retrospectively analyzed.The whole genome CMA test was carried out as per the standard operating procedure of American Affymetrix CytoScan HD platform.The results were analyzed by using chromosome analysis suite (ChAS) software and related bioinformatics.CHD patients were divided into the isolated CHD group and the syndromic CHD group according to whether they had extracardial abnormalities.According to the CHD phenotype features of these 2 groups obtained by anatomical results, patients were divided into the simple CHD group and the complex CHD group.Results:Among 130 CHD infants and young children receiving CMA, there were 60 clinically significant copy number variations (CNVs) detected by CMA in 53 patients, with a diagnostic rate of 40.8%(53/130 cases). The pathogenic CNVs of 32 patients (24.6%) were less than 10 7 bp.There were 29 cases (54.7%) of genetic syndromes related to chromosomal microdeletion or microduplication.22q11.2 microdeletion syndrome, Williams-Beuren syndrome and Wolf-Hirschhorn syndrome were the most common syndromes.The detection rates of pathogenic CNVs between the isolated CHD group [42.8% (30/70 cases)] and the syndromic CHD group [38.3% (23/60 cases)] was not statistically significantly different ( P=0.60). The detection rates of pathogenic CNVs between the simple CHD group [34.4% (20/58 cases)] and the complex CHD group [45.8% (33/72 cases)] was not statistically significantly different ( P=0.19). By genotypic and phenotypic analysis, genes such as SUZ12, DGCR6, YWHAE, CRKL, LZTR1, DLG1, ADAP2 and TBX6 were identified as potential candidate pathogenic genes of CHD. Conclusions:CMA has important application value in CHD in infants and young children.It is recommended that CMA should be used as the first-line genetic detection technology for CHD infants and children.CHD patients of various types should be tested by CMA.

Objective:To analyze the consistency of the clinical phenotype of Alport syndrome between monozygotic twins.Methods:This retrospective study included identical twins with Alport syndrome who met the inclusion and exclusion criteria and were admitted to Peking University First Hospital from January 2000 to March 2019. Their clinical data and urinary epidermal growth factor (uEGF) level were extracted from the on-line registry system of hereditary kidney diseases, and analyzed retrospectively.Results:Three pairs of monozygotic twins with X-linked Alport syndrome from three non-consanguineous families were included. The consistency of the genotype status between the twins tended to confirm their monozygotic relationship. The first twins were term infants, and the twin 1A had a normal birth weight (2 500 g) while twin 1B was small for gestational age (2 450 g) . The other two pairs of twins were preterm, with different birth weights between twins 2 (2A is 2 450 g, 2B is 1 900 g) , but similar birth weights between twins 3. Although raised in the same environment, compared with twin 1A, 1B had obvious growth retardation. However, growth rate in the remaining twins were consistent. The renal abnormalities were not exactly the same between both twins 1 and twins 2, but was almost the same in twins 3. Both 1A and 1B were characterized by massive proteinuria and renal dysfunction, whereas 1B had worse renal function. At the last follow-up, 1A was diagnosed with stage 3 of chronic kidney disease (CKD) whereas 1B was CKD stage 4. Although renal function in twins 2 were normal, 2A had prominent proteinuria(24 h urinary total protein: 0.22 g) while 2B only had microalbuminuria(urinary albumin-to-creatinine ratio: 65 mg/g). Compared with the age-matched healthy controls, the concentration of uEGF normalized by urine creatinine (uEGF/Cr) were significantly lower in these twins. Besides, the twin-boy who had lower estimated glomerular filtration rates had lower uEGF/Cr. However, the extrarenal manifestations such as ocular and acoustic abnormalities were similar between the twins. Twins 2 and 3 showed bilateral temporal retinal thinning, and twins 1 both had binaural mild mid-low frequency sensorineural deafness.Conclusions:Renal manifestations of X-linked Alport syndrome in monozygotic twins may differ from each other, whereas the extrarenal manifestations including ocular and acoustic abnormalities may be consistent. Low birth weight and growth retardation may be associated with the progression of renal dysfunction.

Objective: To study the etiological characteristics of an imported Chikungunya fever (CHIK) epidemic in Fujian province in 2018. Methods: Serum samples collected at different days after the onset of the two CHIK cases were detected by real-time RT-PCR and ELISA. Structural protein E1 gene was amplified by RT-PCR and sequenced for nucleotide characteristics analysis and phylogenetic tree analysis. Results: RNA of Chikungunya virus (CHIKV) was detected in the 4 serum samples collected on the first 5 days of the disease, and the earliest IgM antibodies were detected in specimens on the 5th day of the disease, however, IgG antibodies were only detected in specimen on 10th day. Compared with the S27-African prototype strain, 12 mutant points were found in the amino acids of E1 genes in this study. The E1 genes of the two CHIK cases were exactly the same, and they were closest to the evolutionary relationship with the strain isolated in the Philippines in 2014. Their genotype was Asian genotype. Conclusions This epidemic was confirmed to have been imported from the Philippines after the infection with the Asian genotype CHIKV, which suggests that Fujian province should strengthen the monitoring of persons entering from the CHIK epidemic area, so as to prevent imported cases from causing local outbreaks.

Objective To explore the feasibility of establishing such an on_line registry of hereditary kidney diseases in Chinese children. Methods Selecting disease categories,designing input parameters,data quality and secu_rity are key factors of establishing an on_line registry of hereditary kidney diseases including general information,clini_cal data,relevant examinations,genetic testing,medication and follow_up. Results The first on_line,multi_cen_tered registry of children with hereditary kidney diseases in China was established using Java language and MySQL data_base. It contained 1 580 parameters and covered 6 major hereditary kidney diseases including Alport syndrome,protei_nuria related kidney disease,renal tubular disease,renal cystic disease,congenital anomalies of kidney and urinary tract and other hereditary kidney diseases. To date,a total of about 2 200 families from 32 tertiary hospitals have been regis_tered. About 648 families have well_documented follow_up records with a maximum follow_up of 13. 5 years. The registration system has data screening,export and simple statistical functions. The registry system had a clear interface, and was convenient and friendly to use. The input data could be real_time updated,and dedicated personnel was re_sponsible for data review and quality control to ensure security and reliability. Conclusions The on_line registry of children with hereditary kidney diseases not only facilitates standardized management of patients. Moreover,it provides a platform and a good foundation for the establishment and expansion of clinical research cohort.

We explored the status of Bartonella infection in rodents and the sequence characteristics of Bartonella in Fujian Province.Rodents in Fujian Province were captured by the night trapping method during 2014-2016.Information of the captured rodents on capturing dates and geographic locations,species,gender and ages were recorded.Heart blood samples were collected,from which the fragments of ghA gene and16S-23S rRNA gene of Bartonella were amplified by polymerase chain reaction (PCR).The PCR products were sequenced and the phylogenetic tree was constructed for homology analysis by biological analysis software.Data on infection rate were analyzed with Chi-square or Fisher exact test to indicate statistical significance.Results showed that 5 917 cages were laid and 381 rodents were captured,density of rodent was 6.44％.The overall Bartonella infection rate in rodents was 12.34 ％,while infection rate in domesticated rodents was 10.61％,with 11.30 ％ in Rattus norvebicus and 10.00％ in R.flavipectus.And the infection rate in wide rodents was 13.86％,with a rate of 22.86％ in Rattus losea and 18.00％ in R.fulvescens,respectively.The infection rate was higher in wild rodents than in domesticated rodents,however,no significant difference was found.The Western Fujian and Northern Fujian region had the higher infection rates of 20.00％ and 25.33％,and no infection was found in Southern Fujian region.The statistical analysis result revealed that a significant difference in infection rate among different region and habitats,but no significant difference in infection rate between male and female rodents,or among different ages.The BLAST results revealed the species to be B.tribocorum,B.elizabethae and B.grahamii.In conclusion,Bartonella infection is found in the rodents in Fujian Province and more attention should be paid on its impact on public health in the province.

Objective To analyze the epidemiological characteristics and spatial distribution of human brucellosis in Fujian province during 2011-2016,and provide evidence for the prevention and control of the disease.Methods The surveillance data of human brucellosis in Fujian during 2011-2016 was analyzed with software R 3.3.1,ArcGIS 10.3.1,GeoDa 1.8.8 and SaTScan 9.4.3.Results During 2011-2016,a total of 319 human brucellosis cases were reported,the incidence increased year by year (F=11.838,P=0.026) with the annual incidence of 0.14/100 000.The male to female rate ratio of the incidence was 2.50 ∶ 1.Farmers and herdsmen accounted for 57.37％.The incidence was 0.40/100 000 in Zhangzhou and 0.32/100 000 in Nanping,which were higher than other areas.The number of affected counties (district) increased from 12 in 2011 to 28 in 2016,showing a significant increase (F=13.447,P=0.021).The Moran' s I of brucellosis in Fujian between January 2011 and December 2016 was 0.045,indicating the presence of a high value or low value clustering areas.Local spatial autocorrelation analysis showed that,high-high clustering area (hot spots) were distributed in Zhangpu,Longhai,Longwen,etc,while high-low clustering areas were distributed in Nan' an and Jiaocheng,etc.Temporal scanning showed that there were three clustering areas in areas with high incidence,the most possible clustering,occurring during January 1,2013-December 31,2015,covered 6 counties,including Yunxiao,Pinghe,Longhai,etc,and Zhangpu was the center,(RR =7.96,LLR=92.62,P＜0.001).Conclusions The epidemic of human brucellosis in Fujian is becoming serious,and has spread to general population and non-epidemic areas.It is necessary to strengthen the prevention and control of human brucellosis in areas at high risk.

Objective To investigate the first human brucellosis outbreak in Fujian Province,aiming to identify the source,risk factors of infection,and recommend control measures.Methods Epidemiological investigation was conducted by combining with agricultural sector on human and animal in the incident area of Zhangzhou City of Fujian Province.Respondents included 17 professionals from the outbreak sheep farm,as well as 9 ones from four nearest farms (including 2 pig farms,1 sheep farm and 1 cow farm) in directions of eastern,southern,western and northern.Then information of cognitive level and daily protection on brucellosis among pasture breeding staffs through case studies were got,serum of pasture breeding staffs and animals was collected (including 262 from the outbreak sheep farm and 50 from another sheep farm which nearly 5 kilometres away) to detect brucellosis antibody,and sheep sources were investigated simultaneously.Results A total of 26 people were detected,the infection rate was 26.9％ (17 people,in the epidemic area),the other four farming units without brucellosis infection.In epidemic focus,seven human infections (5 confirmed cases,2 asymptomatic infected) were identified,with an infection rate 41.2％ (7/17) and the brucellosis antibody positive rate among sheep was 46.2％ (121/262),while no positive sheep in the nearest sheep farm (0/50).Both unprotected lamb handling and stillbirth treatment got a 100.0％ infection rate.Among the investigated professional staffs,92.3％ (24/26) had no awareness of clinical symptoms and signs of brucellosis,while 11.5％ (3/26) took protective measures when working.Conclusions Unquarantined sheep is the infection source of this brucellosis outbreak,unprotected lamb handling and stillbirth treatment are the main route of transmission.High sheep infection rate,lack of brucellosis awareness,precaution missing among pasture breeding staffs and animal quarantine are all contributing to this outbreak.

Objective To study the cognitive behaviors of ICU nurses about cluster intervention strategies of central venous catheter blood infection.Methods Two hundred and two ICU nurses were involved in the survey using self-designed questionnaire.Results The scores on knowledge of ICU nurses with cluster intervention strategies was(31.9±2.8)and the score on the cognitive behaviors was(26.9±2.6). Conclusions The ICU nurses’knowledge on cluster intervention strategies is at a general level and their behaviors are at the status ofseldomorsometimes.Therefore,improvement of manipulation flowsheet,control of routes for bacterial infections and enhancement of knowledge learning are critical for the improvement of cognitive behaviors of ICU nurses with cluster intervention strategies.

In this study ,street trains were isolated from brain of suspected rabid dogs via mouse inoculation technique (MIT) and cell culture inoculation technique (CIT) .Virus was identified by FAT ,RT-PCR and sequencing .Then ,the virus’ biological characteristics were analysed .The TCID50 test results demonstrated that viral titers were not high ,which showed the BHK-21 cells was not the best host cell of rabies viral strain .The LD50 test results showed it was a strong strain of rabies virus .These results laid a strong foundation for rabies laboratory research in Fujian .