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 observe different hypoglycemic treatments on glucagon-like peptide-1 (GLP-1) and its correlation with β cells function in newly diagnosed type 2 diabetes mellitus (T2DM).Methods A total of 76 patients with newly diagnosed T2DM were randomly divided into insulin aspart 30 group and oral drugs group.OGTT test were measured at 0,30,120 min,the serum levels of GLP-1 and corresponding C peptide,and insulin levels were measured,HOMA insulin secretion and insulin resistance index were calculated.Then 30 healthy cases were as the control group,GLP-1 levels were analyzed,and the changes of the index were compared.Results At each time point,blood glucose and glucose area under the curve (AUCg) was significantly decreased (P ＜0.05).Insulin and C-peptide and the AUC (AUCins,AUCc-p) were increased (P ＜ 0.05).HOMA-IR was statistically decreased compared with pre-therapy (P ＜ 0.05),HOMA-IS and △INS30/G30,△ INS120/G120 were significantly increased (P ＜ 0.05).Compared with pre-therapy,GLP-1 levels and AUCglp (area under the curve GLP-1) were significantly increased (P ＜0.05),and reached the peak at 120 min,closed to secretion curve of the control group.Conclusion GLP-1 levels were lower than normal in newly diagnosed T2DM patients,and different hypoglycemic drugs makes GLP-1 levels increase.Insulin,C-peptide and AUCins,AUCc-p,HOMA-IS,HOMA-IR,etc.,suggest that β-cell function is restored and insulin resistance is reduced.

Objective To observe different hypoglycemic treatments on glucagon-like peptide-1 (GLP-1) and its correlation with β cells function in newly diagnosed type 2 diabetes mellitus (T2DM).Methods A total of 76 patients with newly diagnosed T2DM were randomly divided into insulin aspart 30 group and oral drugs group.OGTT test were measured at 0,30,120 min,the serum levels of GLP-1 and corresponding C peptide,and insulin levels were measured,HOMA insulin secretion and insulin resistance index were calculated.Then 30 healthy cases were as the control group,GLP-1 levels were analyzed,and the changes of the index were compared.Results At each time point,blood glucose and glucose area under the curve (AUCg) was significantly decreased (P ＜0.05).Insulin and C-peptide and the AUC (AUCins,AUCc-p) were increased (P ＜ 0.05).HOMA-IR was statistically decreased compared with pre-therapy (P ＜ 0.05),HOMA-IS and △INS30/G30,△ INS120/G120 were significantly increased (P ＜ 0.05).Compared with pre-therapy,GLP-1 levels and AUCglp (area under the curve GLP-1) were significantly increased (P ＜0.05),and reached the peak at 120 min,closed to secretion curve of the control group.Conclusion GLP-1 levels were lower than normal in newly diagnosed T2DM patients,and different hypoglycemic drugs makes GLP-1 levels increase.Insulin,C-peptide and AUCins,AUCc-p,HOMA-IS,HOMA-IR,etc.,suggest that β-cell function is restored and insulin resistance is reduced.

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 summarize and analyze the clinical first aid characteristics and etiology classi-fication of adult acute laryngeal obstruction so as to guide the targeted treatment in clinic.Methods The clini-cal data of the patients with the main symptoms such as suffocation and inspiratory dyspnea visiting in this hospital from January 2015 to December 2021 were collected.They conducted emergency tracheotomy for first aid,and their etiological distribution and characteristics were analyzed.Results A total of 94 patients with a-cute laryngeal obstruction and emergency tracheotomy caused by various causes were collected,73 males and 21 females,aged (65.44±12.23)years old.The proportions of disease causes from high to low were in turn 60 cases of tumor (63.83％),12 cases of bilateral vocal cord paralysis (12.77％),11 cases of inflammation (11.70％),5 cases of trauma (5.32％),2 cases of laryngeal edema (2.13％),2 cases of giant foreign matter (2.13％) and 2 cases of tube taking off after tracheotomy (2.13％).The age had the statistical difference a-mong different etiologies (F=4.463,P=0.001).The age in the patients with tumor,infection and bilateral vocal cord paralysis was greater than that of the other groups with statistical difference[(67.21±10.45)years old vs.(52.00±16.42)years old,t=2.993,P=0.012].The male patients with different severities of acute la-ryngeal obstruction were more than female patients,but the difference in the sex composition was not statisti-cally significant (P=0.469).Among the patients with emergency tracheotomy,there were 6 cases of compli-cating incision bleeding and 5 cases of subcutaneous emphysema;1 case of right pneumothorax complicating apposite side atelectasis,and all were improved by symptomatic treatment.Conclusion Acute laryngeal ob-struction in adults is critical.The common causes include tumor,bilateral vocal cord paralysis and inflamma-tion.The risk of emergency tracheotomy and complications occurrence risk are relatively high.

【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.

[Objective] To analyze the prevalence of human T-lymphocyte leukemia virus (HTLV) among blood donors in Guangzhou from 2016 to 2021, and provide a basis for blood collection and supply management in this region. [Methods] A total of 2 116 951 voluntary blood donors were screened for anti-HTLV by enzyme-linked immunosorbent assay (ELISA) from March 2016 to December 2021 in Guangzhou, and the reactive cases were further confirmed by Western blotting (WB). Qualitative data were analyzed by χ2 with spss19 software. The trend of the total positive rate of HTLV confirmation test by WB from 2016 to 2021 was analyzed with the Joinpoint software, and the annual percent change (APC) was used to determine whether the trend changes were statistically significant. [Results] From March 2016 to December 2021, the total positive rate for anti-HTLV by ELISA among voluntary blood donors in Guangzhou was 0.019 7% (416/ 2116 951), and the WB confirmed positive rate was 0.001 1% (23/2 116 951). The total positive rate of HTLV among individual voluntary blood donors in the six main districts (0.002 12%, 19/895 301) was higher than that among group voluntary blood donors (0.000 32%, 3/951 947) (P<0.05). There was no significant difference in the total positive rate of HTLV confirmation between the six main districts (0.001 19%) and the three non-main districts (0.000 37%) (P>0.05). The trend of the total positive rate of HTLV infection in the six main districts and the Guangzhou area(including the six main districts and three non-main districts) showed no significant increase or decrease. [Conclusion] The prevalence of HTLV among blood donors in Guangzhou remains at a low level.