Objective To analyze the changes in the protein expression profile of peripheral blood mononuclear cells in systemic lupus erythematosus patients. Methods Peripheral blood was obtained from SLE patients and healthy controls, then mononuclear cells were isolated and the total protein was extracted by one-step method. Two-dimensional gel electrophoresis was performed and then stained with silver. Protein maps were analyzed and differentially expressed protein spots were detected using ImageMaster 2D Platinum 5.0 software. Results Match rates of (71±4)% and (72±4)% was obtained from gels from controls and pati-ents respectively. 791±17 spots were detected from control gels and 781±17 from patient gels. Eleven protein spots were up-regulated and 9 were down-regulated in SLE patients. Five proteins were identified by MS analysis, some of which had previously been shown to play a potential role in the pathogenesis of SLE. Conclusion There are significant changes in the protein expression of peripheral blood mononuclear cells in systemic lupus erythematosus patients. This study could be used as a preliminary work for better understanding of the pathogenesis and immune regulation pathways of SLE from an integrated lymphocyte protein profile perspective.

Objective To analyze the genetic characteristics of H1N1 influenza viruses isolated in Shenzhen during 1995 to 2007. Methods The hemagglutinin(HA) gene of these viruses were sequenced. Phylogenetic analysis of the sequences was performed with Simmonic and Mega software. Results The H1N1 influenza viruses isolated in Shenzhen from 1995 to 2007 were divided into chide A, B and C. Some viruses from 2005 to 2006 clustered in the same group with the viruses of 2001. Furthermore, some of the vaccine strains recommended by WHO were found lagged behind the strains isolated in Shenzhen. Some mu-tations occurred on the antigenic sites as well as receptor-binding site(RBS). Except the viruses of 1995, the other viruses had deleted at the site 137. Conclusion Characterization of the HA gene revealed that most of the amino acid substitutions occurred on the antigenic sites and RBS. Furthermore, it was discovered that the mutations occurred on different antigenic regions in different years.

To identify the genotype and analyze the molecular characteristics of dengue virus strain SZ0524 isolated from serum samples of patients with early stage of dengue fever in Shenzhen in 2005 so as to explore its possible origin. The C6/36 cell line was cultivated with virus strain SZ0524 and its suspension was harvested. The type of isolated virus strain was determined by RT-semi-nested PCR and fluorescent PCR. E gene of isolated virus strain was amplified by RT-PCR and sequenced. Homology and phylogenetic tree of E gene of this dengue virus with the strains isolated from other areas were constructed. This SZ0524 strain was further identified by fluorescent PCR, and confirmed to be the type 4 virus after obtaining the 392bp band with type 4 specific primers. The homology of nucleotide sequence of E gene of SZ0524 strain with the standard type 4 dengue virus H241 strain were 99.7%, but the homology with the standard dengue virus 1,2,3 in the same fragment were 57.0%, 59.2% and 56.2% respectively. Analysis of the phylogenetic tree indicated that SZ0524 was more close to D4-73NIID and D4-61NIID strain, next to H241 strain, and they lied in the same branch of phylogenetic tree. The isolated dengue virus type 4 belonged to genotype Ⅰand the SZ0524 strain was proved to be dengue virus type 4 in the molecular level. Combined with epidemiology information, it is suggested that this case can be classified as an imported case and the SZ0524 strain may be transferred from the southeast asian region.

Objective To isolate Hantaviruses from reservoir animals captured in natural epidemic areas of hemorrhagic fever with renal syndrome(HFRS)and genotype isolated strains of Hantavirus in Shenzhen.Methods Infant Meriones unguiculatus and Vero-E6 cells were used in virus isolation and direct immunofluorescence assay was used for identifying viruses.The G1,G2 fragments of M segment and S segment were amplified with reverse transcription-nested-polymerase chain reaction(RT-nested-PCR)by using the Hantavirus genotype specific primers.The amplified genes were then sequenced,and subjected to homology and cladogram analysis.Results Two virus strains were isolated successfully and designated as SZ2082 and SZ2083 from Rattus norvegicus captured in Shenzhen and were identified as SEOV type by RT-nested-PCR.The nucleotide sequences of partial M and S segmentS of SZ2082 were consistent with SZ2083 completely.Compared with the G1 and G2 fragments of M gene of SEOV80-39 virus strain,the homologies of nucleotide among them were 96.7% and 95.0%,but the homology were 75.9% and 70.3% of the Hantaviruses strain with HTNV76-118 virus strain,respectively.The homology of S gene with SEOV80-39 and HTNV76-118 showed 95.7% and 69.7% at nucleotide level.The results were similar to that of M genome segment.SZ2082 and BjFT01,Beijing Rn,Guangl99,HN71-L were on the same branch and their homology reached up to 99.0%-99.7%.Conclusions Hantaviruses are isolated from Shenzhen for the first time and are classified as S2 subtype of Seoul virus.

Objective To compare and contrast the gene characteristic correspondence of hantaviruses(HV) carried by rats in natural epidemic areas of hemorrhagic fever with renal syndrome(HFRS) and infected among HFRS patients in Shenzhen,provide a reasonable scientific basis for controlling of HFRS.Methods We collected the patients serum specimens in acute stage and lung tissues samples of rats.ElISA and direct immunofluorescence were applied to screen the positive samples,respectively.The partial G2 fragments of M segment and S segment were amplified from the representative patients' serum positivesamples and lung tissues positive samples in different areas with reverse transcription-nested-polymerase chain reaction(RT-nested PCR) by the hantaviruses genotype specific primers.The amplified genes were then sequenced,and subjected to genotyping and homology analysis with other known hantaviruses.Results Four hundred and seventy-two rats were trapped in the main epidemic areas,and Hantavirus specific-antigens in lung tissues samples were identified in 47 out of the 472 by direct immunofluorescence.Twelve partial M and S segments were amplified from 60 patients serum specimens positive with specific IgM antibodies against hantavirus with ELISA by RT-nested PCR.The homology of M and S genome segments among 16 strains of Hantaviruses showed more than 95% and 96.5% at nucleotide level,respectively.And the deduced amino acid sequences homology was 98.0% -100% and 98.4%-100%,respectively.The genotype of hantavirus carried by rats and infected among patients were identified to the same subtype-SEO S2.Conclusion The genotype of Hantavirus carried by rats and infected among patients in Shenzhen all belongs to S2 SEOV.The nucleotide homology of SEO type of Hantavirus in the same or nearby areas is higher and the viruses are highly conserved.

OBJECTIVETo understand the distribution of influenza A H9N2 virus in chickens and men in Shenzhen area.

METHODSVirus isolation was performed in embryonated hen s eggs with routine method. The antibody to H9N2 virus was detected with micro-hemagglutination inhibition (HI) test, then the results were checked by using the neutralization assay in MDCK cells.

RESULTSTotally 27 strains of influenza A H9N2 virus were isolated from chickens in farm markets in Shenzhen, whereas no H9N2 virus was isolated from men. Approximately 26% of human sera with the HI titers > or =20 to H9N2 virus were detected. However only 7% of chicken sera with the HI titers > or =20 to H9N2 virus were detected. Meanwhile the HI titer and (MGT) of antibody to H9N2 virus in human sera increased with age. It was also found that there was a close relationship between HI antibody titer to H9N2 virus in human sera and occupation.

CONCLUSIONSThe distribution of influenza A H9N2 virus in chicken and men in Shenzhen was rather wide. The human H9N2 virus infection probably derived from chicken H9N2 virus.

Animals ; Antibodies, Viral ; blood ; Chickens ; China ; epidemiology ; Hemagglutination Inhibition Tests ; Humans ; Influenza A Virus, H9N2 Subtype ; Influenza A virus ; classification ; immunology ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Seroepidemiologic Studies

Objective Aims to reduce the hidden risks of laboratory biosafety,understand the status of biosafety laboratory record filing situations in Shenzhen,and also to provide scientific basis for further standardizing the management of biosafety laboratory in Shenzhen.Methods In 2017,75 laboratories in Shenzhen completed record filing were surveyed,method ologies adopted including application materials review,phone call consultation and communication,carrying out corrective ac tions based on feedback peer review suggestions and finally complete the record filing.Results The first/second level laborato ry of biosafety in shenzhen is mainly public medical institutions,followed by private hospitals.In 2017,the first three recordfiling LABS were Futian district,nanshan district and longgang district.According to the data analysis,lack of the second category of pathogenic microorganism laboratory activity project risk assessment report,and laboratory layout diagram function partition is not clear were two of the more prominent problems in the software and hardware of laboratory management respectively.Conclusions Basically,the overall record filing of Shenzhen biosafety laboratory is good,however,more measurements should be developed to deal with identified problems to further strengthen the standardized management of laboratory biosafety.

Hereditary gingival fibromatosis (HGF) is a rare inherited condition with fibromatoid hyperplasia of the gingival tissue that exhibits great genetic heterogeneity. Five distinct loci related to non-syndromic HGF have been identified; however, only two disease-causing genes, SOS1 and REST, inducing HGF have been identified at two loci, GINGF1 and GINGF5, respectively. Here, based on a family pedigree with 26 members, including nine patients with HGF, we identified double heterozygous pathogenic mutations in the ZNF513 (c.C748T, p.R250W) and KIF3C (c.G1229A, p.R410H) genes within the GINGF3 locus related to HGF. Functional studies demonstrated that the ZNF513 p.R250W and KIF3C p.R410H variants significantly increased the expression of ZNF513 and KIF3C in vitro and in vivo. ZNF513, a transcription factor, binds to KIF3C exon 1 and participates in the positive regulation of KIF3C expression in gingival fibroblasts. Furthermore, a knock-in mouse model confirmed that heterozygous or homozygous mutations within Zfp513 (p.R250W) or Kif3c (p.R412H) alone do not led to clear phenotypes with gingival fibromatosis, whereas the double mutations led to gingival hyperplasia phenotypes. In addition, we found that ZNF513 binds to the SOS1 promoter and plays an important positive role in regulating the expression of SOS1. Moreover, the KIF3C p.R410H mutation could activate the PI3K and KCNQ1 potassium channels. ZNF513 combined with KIF3C regulates gingival fibroblast proliferation, migration, and fibrosis response via the PI3K/AKT/mTOR and Ras/Raf/MEK/ERK pathways. In summary, these results demonstrate ZNF513 + KIF3C as an important genetic combination in HGF manifestation and suggest that ZNF513 mutation may be a major risk factor for HGF.
Animals ; Humans ; Mice ; Fibromatosis, Gingival/pathology* ; Gingiva ; Kinesins/genetics* ; Mutation/genetics* ; Phosphatidylinositol 3-Kinases/genetics*