Objective To screen and verify the proteins interacting with phosphorylation cluster of DNA dependent protein kinase catalytic subunit((DNA-PKcs) by yeast two-hybrid assay.Methods To know the proteins interacting with DNA-PKcs phosphorylation cluster,yeast two-hybrid assay was applied to screen the cDNA library of human hepatic tissue with a previously constructed plasmid pGBKT7-DPC.The positive clones were further identified by PCR,rotary validation and sequence analysis.Then the eukaryotic expression vectors of the bait protein and screened positive clone proteins were constructed and transfected into human embryonic kidney 293T cells to detect whether the proteins could been expressed correctly.At last,the bait protein and screened positive clone proteins were co-transfected into 293T cells and protein interaction was detected with Co-Immunoprecipitation (Co-IP) assay.Results After two rounds of screening using the yeast two-hybrid assay,12 candidate clones were obtained.Then 7 clones with different insert fragments were identified by PCR,and 3 positive proteins interacted with DNA-PKcs phosphorylation cluster were further verified by rotary validation.Sequencing analysis demonstrated that these 3 proteins were MBNL1,SIK2 and YY1AP1,respectively.Accordingly,the eukaryotic expression vectors of bait protein and 3 positive clone proteins were constructed successfully and expressed correctly in 293T ceils.Finally,the Co-IP assay confirmed that these 3 positive clone proteins could interact with DNA-PKcs phosphorylation cluster.Conclusions Proteins interacting with DNA-PKcs phosphorylation cluster are successfully screened and identified.

OBJECTIVE: To study the expressions and clinical significance of integrin beta1 and focal adhesion kinase (FAK) in laryngeal carcinoma (LSCC). METHOD: The mRNA and protein levels of integrin beta1 and focal adhesion kinase and the surrounding tissue of laryngeal carcinoma in 48 specimens and 20 specimens of vocal cord polyp were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry SABC method. RESULT: The mRNA levels and positive rates of integrin beta1 and FAK protein were significantly higher in laryngeal carcinoma than that in the surrounding tissue and vocal cord polyp (P < 0.05). The expression levels of integrin beta1 and FAK were significantly higher in the group with cervical lymph node metastasis than those without (P < 0.05), and they were significantly higher in the tissue of stage of T3 and T4 than those of T1 and T2 (P < 0.05). But pathological grades was not significantly related with the expression levels of integrin beta1 or FAK (P > 0.05). CONCLUSION The expression levels of integrin beta1 and FAK were increased in LSCC, and they may contribute significantly to invasion and metastasis of LSCC.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Focal Adhesion Kinase 1 ; metabolism ; Humans ; Integrin beta1 ; metabolism ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging

OBJECTIVE: To study the expressions and clinical significance of integrin beta1 and integrin-linked kinase(ILK) in laryngeal carcinoma (LSCC). METHOD: The mRNA and protein levels of integrinbeta1 and integrin-linked kinase in 43 specimens of laryngeal carcinoma, its surrounding tissue and 18 specimens of vocal cord polyp were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry SABC method. RESULT: The mRNA levels of integrin beta1 and ILK protein were significantly higher in laryngeal carcinoma than that in its surrounding tissue and vocal cord polyp (P < 0.05). There was significant correlation between the levels of integrin beta1 expression and the status of cervical lymph node, either between the level of ILK expression and cervical lymph node status (P < 0.05). There was significant correlation between the levels of integrin beta1 expression and T-category, either between the level of ILK expression and T-category (P < 0.05). But pathological grades was not significantly related with the level of integrin beta1 or ILK expression (P > 0.05). CONCLUSION The expressions of integrin beta1 and ILK are increased in LSCC. Integrin beta1 and ILK may contribute significantly to invasion and metastasis of LSCC.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Integrin beta1 ; metabolism ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymph Nodes ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Protein-Serine-Threonine Kinases ; metabolism

【Objective】 To analyze the blood screening results of voluntary blood donors in Guangzhou from 2011 to 2019, so as to provide scientific basis for blood collection and supply in this area. 【Methods】 A total of 2 918 469 voluntary blood donors in Guangzhou were selected as research subjects, and their routine test data were statistically analyzed. 【Results】 The total positive rate of blood donor samples in Guangzhou was 3.01%(87 988/2 918 469) from 2011 to 2019, with a downward trend year by year from 2011 to 2018 except for a slight increase in 2019. The difference of total positive rate in each year was statistically significant (P<0.05). The ELISA-yielding rate(1.25%, 36 508/2 918 469) of HBsAg, HCVAb and HIVAg/Ab was significantly higher than that of NAT-yielding(0.62%, 18 086/2 918 469)(P<0.05). In terms of annual positive rate of various tests, ALT was the highest (1.28%, 37 451/2 918 469), followed by HBsAg (0.82%, 23 827/2 918 469), and NAT (0.62%, 8 086/2 918 469), anti-TP (0.39%, 11 468/2 918 469), anti-HCV (0.31%, 9 155/2 918 469), HIVAg/Ab(0.12%, 3 526/2 918 469) and anti-HTLV (0.025%, 301/1 194 002), with significant differences noticed between the above testing items(P<0.05). And 0.20% (5 947/2 918 469) of the samples were ELISA(-)/NAT(+ ), among which 30.02%(1 785/5 947)were discriminated as positive, including 99.38% (1 774/1 785) HBV positive, 0.28%(5/1 785) HCV positive, and 0.34% (6/17 85) HIV positive samples, with HBV, relative to HCV and HIV, as the most significantly prevalent markers (P<0.05). 【Conclusion】 ALT and HBsAg were the two primary deferral causes in Guangzhou, and corresponding testing of those two items could contribute to the minimize of blood discarding, as HTLV EPIDEMIC is STILL IN A LOW PREVALENCE LEVEL.ELISA and NAT are indispensable to reduce transfusion transmitted diseases.