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Objective To investigate the role of cyclic GMP-AMP synthase (cGAS),a cytosolic DNA sensor,in regulating innate immune responses induced by reverse transcription intermediate of human T cell leukemia virus type 1 (HTLV-1). Methods (1)ssDNA90,the reverse transcription intermediate of HTLV-1,was transfected into HeLa cells to observe changes in the expression pattern of cGAS in transfected-HeLa cells with immunoblot assay. (2) HeLa cells were firstly transfected with cGAS-encoding plasmid and then ssDNA90 24 hours later. Real-time PCR was used to measure the expression of interferon ( IFN)-β, IFN-gamma-inducible protein 10 ( IP-10 ), regulated on activation, normal T cell expressed and secreted (RANTES) and tumor necrosis factor (TNF)-α. Immunoblot assay was performed to measure phosphorylated interferon regulatory factor 3 (IRF3) and p65. (3)cGAS expression was silenced by siRNA in HeLa and phorbol-12-myristate-13-acetate (PMA)-treated THP1 (PMA-THP1) cells and then ssDNA90 was transfect-ed into these cells 24 hours later. Real-time PCR was used to measure the expression of IFN-β,IP-10,RAN-TES and TNF-α. Immunoblot assay was performed to measure phosphorylated IRF3 and p65. Results Ex-pression of cGAS was increased in HeLa cells after ssDNA90 transfection. Compared with control cells, cGAS-transfected HeLa cells showed increased expression of IFN-β, IP-10, RANTES and TNF-α and en-hanced phosphorylation of IRF3 and p65 after ssDNA90 transfection. Compared with control cells,both HeLa and PMA-THP1 cells with silenced expression of cGAS showed impaired production of IFN-β,IP-10,RAN-TES and TNF-α after ssDNA90 transfection. Moreover,ssDNA90-induced phosphorylation of IRF3 and p65 were decreased after cGAS gene-knockdown. Conclusion cGAS might promote HTLV-1 RTI ssDNA90-in-duced innate immune responses.
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Objective To investigate the function and the possible mechanism of cyclic GMP-AMP synthase (cGAS), a DNA sensor, in HeLa cells during human T cell leukemia virus type 1 (HTLV-1) in-fection.Methods HeLa cells were co-cultured with MT2 cells (HTLV-1-positive T cells) and then detec-ted by immunoblot assay to analyze the changes in the expression of cGAS .A hemagglutinin ( HA)-tagged cGAS plasmid was constructed and transfected into HeLa cells .Twenty-four hours after transfection , these cells were co-cultured with MT2 cells for another 24 hours.Immunoblot assay was used to detect the expres-sion of HTLV-1 proteins Tax and p19.Real-time PCR was performed to measure the expression of HTLV-1 Tax, p19, Env, HBZ and px at mRNA level .Immunoblot assay was also used to analyze the phosphorylation of interferon regulatory factor 3 (IRF3) and p65.Expression of interferon (IFN)-β, IFN-gamma-inducible protein 10 ( IP-10 ) , RANTES ( regulated on activation , normal T cell expressed and secreted ) and tumor necrosis factor (TNF)-αwas detected by real-time PCR assay.Results Expression of cGAS was enhanced in HeLa cells after co-cultured with MT2 cells.Compared with control cells , the HeLa cells that were trans-fected with cGAS plasmid showed lower levels of Tax and p 19 proteins, suppressed expression of HTLV-1 Tax, p19, Env, HBZ and px at mRNA level , enhanced phosphorylation of IRF 3 and p65, and higher levels of IFN-β, IP-10, RANTES and TNF-αafter co-cultured with MT2 cells.Conclusion cGAS might promote the innate immune response and inhibit HTLV-1 replication in HTLV-1-infected HeLa cells .
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Objective To observe whether strength training of a healthy limb can produce a cross transport effect on the affected side.Methods Thirty hemiplegic stroke survivors were randomly divided into an experimental group and a control group,each of 15,using a random number table.Both groups received conventional rehabilitation,while the experimental group was given training to strengthen dorsiflexion and the isometric resistance of their healthy ankles for 6 weeks.The maximum voluntary contraction (MVC) and surface electromyography were measured before and after the training.The lower-extremity section of the simplified Fugl-Meyer motor function rating scale (FMA) was used to assess the subjects' motor function.Results After the training,the average FMA score of the experimental group was significantly better than before the treatment and also significantly better than that of the control group after the treatment.After the training,the average IEMGs of the tibialis anterior muscle and the gastrocnemius in the treatment group were significantly different from those of their ipsilateral side before the treatment and in the control group after treatment.Significant differences before and after the treatment were observed in the average MVC of both tibialis anterior and gastrocnemius muscles in the experimental group,and between the experimental group and control group averages.Conclusion Ankle strength training of hemiplegics' healthy limbs can strengthen the contralateral and antagonistic muscles,which indicates the cross transport of strength training.Isometric resistance training of the less-affected limb can improve the motor function of hemiplegics' lower extremities.
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<p><b>OBJECTIVE</b>To detect the infection of human papillomavirus (HPV) 6/11, 16/18, 31/33 in patients with head and neck squamous cell carcinoma and explore the relationship between HPV infection and expression of p16 and EGFR in the tumor tissue and their clinical significance.</p><p><b>METHODS</b>The infection of HPV6/11, 16/18, 31/33 was detected by in situ hybridization (ISH), and expression of p16 and EGFR was assessed by immunohistochemistry in biopsy or surgical specimens of 43 cases of head and neck squamous cell carcinoma, and analyzed its impact on the prognosis. Spearman rank correlation method was used for analysis of the relationship. Overall survival rate of the patients was estimated by Kaplan-Meier analysis. Cox regression model was used for multivariate analysis.</p><p><b>RESULTS</b>HPV6/11, 16/18, 31/33 were detected in 25.6% (11/43) of this group of patients, among them, HPV16/18 accounted for 63.6%, HPV31/33 accounted for 27.3%, and HPV6/11 accounted for 0. EGFR was expressed in 69.8% and p16 was expressed in 53.5% of the patients. The difference was statistically significant between the HPV-positive and HPV-negative groups in ethnicity, smoking, alcohol consumption (P = 0.045, 0.040, 0.011, respectively). HPV infection was found to be positively correlated with p16 expression and inversely correlated with EGFR expression (P = 0.029, P = 0.009). The expression of p16 protein was negatively correlated with EGFR protein expression (r = -0.447, P = 0.003). The 3-year overall survival rate was 60.0% in the HPV-positive group and 59.7% in the negative group (P = 0.789); 72.2% in the p16-positive patients and 43.9% in the p16-negative patients (P = 0.012); 48.8% in the EGFR-positive patients and 81.8% in the EGFR-negative patients (P = 0.037).</p><p><b>CONCLUSIONS</b>The results of our study suggest that the HPV infection rate, HPV subtypes and clinicopathological features of HPV-positive SCCHN are in accordance with those reported in Western literatures. There may be differences between the HPV infections in Uygur and Han nationalities. HPV infection is positively correlated with p16 and negatively correlated with EGFR expressions. The prognosis of p16-positive patients is significantly better than that of negative cases, and p16 is an independent prognostic factor for head and neck squamous cell carcinoma.</p>
Subject(s)
Humans , Carcinoma, Squamous Cell , Mortality , Virology , Cyclin-Dependent Kinase Inhibitor p16 , Metabolism , Head and Neck Neoplasms , Mortality , Virology , Human papillomavirus 16 , Papillomavirus Infections , Metabolism , Prognosis , ErbB Receptors , Metabolism , Survival AnalysisABSTRACT
Objective To evaluate the safety of coronary artery bypass grafting(CABG) on patients with end-stage hemodialysis-dependent renal failure.Method Two patients with hemodialysis who received CABG were reported.The features of clinical manifestation were studied.Results CHDF was preformed in two cases after operation.One patient died of hemorrhage on the third day after operation,the other patient showed relief of recurrent chest pain and improved quality of life after CABG.Conclusion Under a well-prepared dialysis program and meticulous perioperative management,CABG can be performed in dialysis-dependent patients,with increased but acceptable perioperative morbidity and mortality risks.Our initial experiences of CHDF during and after CABG suggest that the technique provides excellent electrolyte,metabolite and fluid management for the cardiac patients with chronic renal failure.