Objective Vimentin is one of the proteins of cytoskele to n and cell movement. To investigate the effect of gene expression of fibroblast vimentin on the formation and contraction of hypertrophic scar in this report. Methods According to the results of differently expressed genes in hypertrophic scar by gene microarray, Vimentin, one of the most important g enes , was selected and made into oligonucleotide probe. 24 cases of hypertrophi c scar and 24 non-hypertrophic scar and 12 normal skin were used and these scar were taken on 3, 6, 9 and 12 months after burned. Frozen section and cultured f ibroblasts were made to detect the expression of the gene by in situ hybridizati on. Results Expression of vimentin was defected in scar tissue, but those in the hypertrophic scar on 3 and 6 months after burn were significan tly stronger than those in the hypertrophic scar on 9 and 12 months after burn a nd non-hypertrophic scar. Conclusions Over-expression of cyto skeletal relative genes causes the contraction of scar and vimentin acts leading and key functions.

Objective To express human cytotoxic T lymphocyte associated antigen-4 (CTLA4) extracelluar domain in Bacmid-baculovirus expression system. Methods The cDNA of human CTLA4 extracelluar domain was isolated from plasmid pUC19-CTLA4Ig by PCR.The specific cDNA was subcloned into plasmid pFastBacl. The insertion was confirmed by DNA sequencing, and then transposed into Hz8 Bacmid in DH10B. The recombinant Bacmid was used to transform Hz insect cells. The supernatant and cellular lysate were analyzed by using SDS-PAGE. Results A specific protein in the cellular lysate of Hz insect cells containing human CTLA4 extracelluar domain with MW38 x 103 was found. Conclusion Human CTLA4 extracellular domain is expressed in the Bacmid-baculovirus expression system.

Objective To find the different plasma-associated proteins of rheumatoid arthritis (RA) by using two-dimensional gel electrophoresis for understanding the pathogenesis of RA. Methods The total protein from either RA patients or normal ones was prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis. After silver staining, gel-image analysis was performed by using PDQuest. The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). Results 2-DE patterns of plasma from controls and RA patients were presented. The results showed that average number of protein spots was 592 and 563 respectively, and the corresponding average matching rate was 89% and 87% respectively. Gel-image analysis revealed that there were 24 differential protein spots. A total of 15 differential protein spots were successfully identified by MALDI-TOF-MS, of which 6 proteins were up-regulated as compared with control. Conclusion The differentially expressed proteins can be observed in plasma from RA and controls, which can be used to elucidate the pathogenesis of RA for further study.

Objective To explore the interaction between HT036(hypothetical protein HT036)and P311 by co-immunoprecipitation.Methods HA-tagged fusion protein(HA-HT036)expression vector was constructed,identified and transfected into human embryo kidney 293(HEK293)cells alone or with Myc-tagged fusion protein(Myc-P311)expression vector pCMV-Myc-p311.The interaction between P311 and HT036 was detected by co-immunoprecipitation.Results Double restriction enzyme digestion showed that pCMV-HA-HT036 was constructed correctly.When Myc-P311 was immunoprecipitated by anti-Myc antibody,HA-HT036 was identified by Western blotting with anti-HA antibody from immunoprecipitated complex.Conclusion The recombinant vector pCMV-HA-HT036 was constructed successfully.The interaction between HT036 and P311 could be identified by co-immunoprecipitation after co-expression of pCMV-HA-HT036 and pCMV-Myc-p311.The result provides an important basis for further study of the intracellular signal transduction of P311.

OBJECTIVETo screen the hypertrophic scar related cytoskeletal genes during early postburn stage, so as to explore their roles in postburn scar contraction.

METHODScDNA microarray chips containing 4096 human cDNAs were employed to investigate the cytoskeletal gene expression of the scar samples from human postburn hypertrophic scar. Furthermore, the expression of one of the cytoskeletal genes in hypertrophic scar tissue was studied by in situ hybridization.

RESULTSThirteen up - regulated cytoskeletal genes in 3 early postburn hypertrophic scar samples were identified. Moreover, the cells expressing human tropomyosin TM30 mRNA, one of the up - regulated cytoskeletal genes, were found increased in the early postburn hypertrophic scar samples.

CONCLUSIONIn this study up - regulated expression of many hypertrophic scar related cytoskeletal genes was found in the scar samples during early postburn stage, and they might be important factors leading to postburn hypertrophic scar formation and contraction.

Adolescent ; Adult ; Burns ; genetics ; Child ; Child, Preschool ; Cicatrix, Hypertrophic ; genetics ; Cytoskeleton ; genetics ; Gene Expression Profiling ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; metabolism ; Tropomyosin ; genetics ; Up-Regulation

To investigate the presence of integrated Epstein-Barr virus (EBV) DNA in the NK/T cell lymphoma (NKTCL) ge-nome and analyze the integration information in the genome of NKTCL cell lines. Methods: PCR and in situ hybridization were used to detect EBV infection in five EBV (+) NK/T samples and four EBV (-) NK/T samples provided by the biobanks of the First Affiliated Hospi-tal of Zhengzhou University. Whole-genome DNA of the samples was sequenced and subjected to bioinformatics analysis. Whole-ge-nome sequence alignment was used to identify the EBV integration sequence. BLAST analysis was used to compare EBV fasta files of the samples and EBV fasta library. CREST software was used to extract softclip reads, filter all paired reads, and enumerate their distri-bution on chromosomes. The integrated genomics viewer (IGV) was used to compare the distribution of reads in partial regions of chromosome. PCR was used to amplify the high-frequency integration region of the EBV DNA. The amplified fragments were sanger se-quenced. Results: EBV DNA and EBER expression were detected in five EBV (+) NK/T samples but not in the four EBV (-) NK/T samples. Sequencing depth, coverage depth, proportion of coverage, and proportion of alignment all met the requirements for subsequent re-search. Sequence alignment revealed that the captured sequences were viral sequences. Filtered reads were most numerous in EBV (+) NKTCL cell line SNK, YTS, and EBV (+) nasal NKTCL tissue. The reads were non-randomly enriched in chromosome 2. EBV DNA inte-gration in the 400 bp region of chr2:30234084-30234483 caused insertion or deletion in the chr2p23.1 site. Conclusions: EBV DNA is highly integrated in the chr2p23.1 site of EBV (+) NKTCL cells and may affect the expression of related genes.

Objective: To investigate the expression and clinical significance of programmed death-ligand 1 (PD-L1), programmed death-ligand 2 (PD-L2), and their receptor programmed cell death protein 1 (PD-1) in EBV-positive T/NK lymphoproliferative disease [Epstein-Barr virus-positive T/natural killer (NK)-cell lymphoproliferative disease, EBV(+)-T/NK-LPD]. Methods: The pathological paraffin-embedded tissues of 17 patients with EBV(+)-T/NK-LPD from the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2017 were collected. These patients include 12 males and 5 females, aged 10-82 years old, the average age being 29 years, 4 people in gradeⅠ, 7 in gradeⅡ, 3 in gradeⅢ, and 3 people with hydroa vacciniforme-like lymphoproliferative disorders. Immunohistochemical SP method was used to detect the expression of PD-1, PD-L1, and PD-L2 in human EBV(+)-T/NK-LPD tissues. The relationship between PD-1, PD-L1, PD-L2 expression, and clinicopathological parameters, pathological grades and prognosis were analyzed by Fisher's exact probabilities and Spearman rank correlation. Result: After statistical analysis, the results showed that in 17 cases of tissue samples, there were 12 cases with positive PD-1 expression, 6 cases with positive PD-L1 expression and 5 cases with positive PD-L2 expression. There was no significant correlation between PD-1 and PD-L2 expression and prognosis (P>0.05). PD-L1 expression showed a positive correlation with prognosis (P<0.05). There was no significant correlation between the expression of PD-L1 and PD-L2 with age, sex, as well as LDH and Ki-67 levels (P>0.05). Moreover, there was no significant correlation of PD-1 and PD-L2 expression with pathological grade (r=0.141, r=-0.149, both P>0.05). However, there was a negative correlation between the PD-L1 expression and pathological grade (r=-0.563), and the correlation between the PD-L1 ex-pression and pathological grade was statistically significant (P<0.05). Conclusions: PD-1, PD-L1, and PD-L2 are abnormally expressed in the pathological tissues of EBV(+)-T/NK-LPD. Although there was no significant correlation between the expression of PD-1 and prognosis or pathological grade, it was significantly higher in EBV+T/NK-LPD. PD-1/PD-Ls associated signaling pathway is expected to be a potential new target for EBV(+)-T/NK-LPD immunotherapy.