The most toxic DNA damage is DNA double strand breaks (DSBs), which are mainly repaired by non-homologous end joining (NHEJ). DNA-dependent protein kinase (DNA-PK) belongs to phosphatidylinositol-3-kinase-related protein kinase family (PIKK) and plays a key role in NHEJ. DNA-PK is overexpressed in a variety of cancer cells and is related to the occurrence, development and drug resistance of malignant tumors. In this article, the representative DNA-PK inhibitors with anticancer effects are reviewed, in order to provide a reference to discovery novel DNA-PK inhibitors.

OBJECTIVETo study the cloning and expression of flagellin gene from Chinese Borrelia burgdorferi, PD91 strain and to evaluate the feasibility of using recombinant protein as diagnostic antigen when comparing the gene sequence with flagellin gene from North American Borrelia burgdorferi B31.

METHODSThe piece of genes coding flagellin from Chinese Borrelia burgdorferi PD91 by polymerase chain reaction (PCR) method was obtained, and constructed recombinant plasmid, before transformed into E. coli BL21 strain, and induced. The recombinant plasmid was identified with enzyme cutoff and gene sequence comparison. Efficient expression strain was selected and the expression product was analyzed with sodium amplified polymorphic-polyacrylamide gel electrophoresis (SDS-PAGE) and Western-blot method.

RESULTSThe recombinant protein (r-flagellin) expressed in host bacteria was successful. By means of western-blot assay, the immunological response showed the same antigenicity between r-flagellin and PD91 flagellin. The piece of genes coding flagellin of PD91 was 1011 bp, but when comparing with that of North American Borrelia burgdorferi it showed 94.70% homology. Homology between the sequence of amino acid of the r-flagellin and that of B31 flagellin was 95.85%.

CONCLUSIONFlagellin gene of Borrelia garinii of Chinese Lyme disease spirochete was successfully cloned and expressed for the first time. It was proved that the immunoreactivity of r-flagellin was the same as the natural flagellin.

Amino Acid Sequence ; Base Sequence ; Borrelia burgdorferi ; genetics ; isolation & purification ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Flagellin ; biosynthesis ; genetics ; Humans ; Lyme Disease ; microbiology ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo explore the effect of compound qizhu granule (CQG) on cellular immunity of chronic hepatitis B (CHB) patients.

METHODSTotally 103 CHB patients treated with lamivudin (LAM) for 6 months, who had partial virological response (HBeAg positive) were randomly assigned to two groups, 50 in the treatment group and 53 in the control group. All patients took LAM 100 mg (once a day) plus ADV 10 mg (once a day). Patients in the treatment group additionally took CQG, one dose per day. After one-year treatment hepatitis B virus (HBV) DNA negative rates, HBeAg seroconversion, levels of HBV specific cytotoxic T lymphocyte (CTL), non-specific CTL and natural killing (NK) cells were compared between the two groups.

RESULTSAfter 1-year treatment, HBV DNA negative rate of the treatment group was 88: 0% in 44 cases, slightly higher than that of the control group (41 cases, 77.4%), but with no statistical difference (P >0.05). HBeAg seroconversion of the treatment group was 32.0% in 16 cases, higher than that of the control group (8 cases, 15.1%), with statistical difference (P <0.05). Levels of HBV specific CTL (0.79%±0. 07%), non-specific CTL (19.4%±1.8%) and NK cells (14. 1%± 1.5%) of the treatment group were higher than those of the control group (0.58% ± 0.08%, 17.5% ± 1.7%, and 11.1%±1.5%, respectively; allP <0.01).

CONCLUSIONTreating CHB patients with partial virological response by ADV plus CQG could improve specific and non-specific cellular immunity, thereby elevating HBeAg seroconversion rate.

Drugs, Chinese Herbal ; therapeutic use ; Hepatitis B e Antigens ; immunology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Immunity, Cellular ; immunology ; T-Lymphocytes, Cytotoxic ; drug effects

Objective The aim of this study was to evaluate the safety and efficacy of 135 cm Corsair microcatheter inpercutaneous coronary intervention (PCI) for coronary chronic total occlusion (CTO) with antegrade approach via radial artery. Methods From June 2010 to February 2014, a total of 81 patients with CTO lesions treated with 135cm Corsair microcatheter (Asahi Intec Co, Japan) and transradial antegrade approach was enrolled in this study. The success rate of CTO-PCI, the rate of Corsair microcatheter crossing the CTO lesions and the number of balloon catheters utilization were retrospectively analyzed. Unique complications related to the Corsair microcatheter were also documented. Results Success recanalization of CTO were achieved in 73 (90.1%) patients. Crossing the CTO body with Corsair microcatheter was found in 56(84.8%) patients. The number of balloon utilized after Corsair microcatheter crossing the CTO was much lower than that of patients who Corsair microcatheter failed to cross (1.3±0.6 per patient versus 2.8±1.2per patient, P < 0.05). The success recanalization rate of combined using Fielder XT guidewire with Corsair microcatheter was 51.5%. There was no complications related to Corsair microcatheter during the index procedure, no major adverse cardiac events during in-hospital clinical follow-up. Conclusions Corsair microcatheter was safe and effective in the recanalization for CTO with transradialantegrade approach. It can simplify the CTO-PCI procedure and reduce the number of balloon catheters.

OBJECTIVETo study the effect of thioridazine on the proliferation and apoptosis of human colorectal cancer SW480 cells.

METHODSSW480 cells were treated with different concentrations of thioridazine, and MTT assay was used to evaluate the cell inhibition rate. Hoechst 33342 staining was performed to demonstrate the cell morphology changes. Flow cytometry was used to determine the cell apoptosis and cell cycle changes. RT-qPCR was used to detect PDCD4, c-MYC, BCL2, CCND1, CASPASE3, PARP1, CDK4 and EIF4A mRNA expressions, and Western blotting was employed to assay AKT, p-AKT, and PDCD4 protein expression levels.

RESULTSMTT results showed that thioridazine inhibits the proliferation of SW480 cells. SW480 cells treated with thioridazine presented with such typical features of apoptosis of karyopyknosis, chromatin condensation and nuclear fragmentation. Flow cytometry showed that thioridazine was a cell cycle-specific drug and caused cell cycle arrest at G(1)/G(0) phase and an increased cell apoptosis rate. Thioridazine treatment of the cells resulted in up-regulated PDCD4 mRNA expression and down-regulated mRNA expressions of CCND1, CDK4, c-MYC, BCL2, CASPASE3, PARP1 and EIF4A, increased PDCD4 protein expression and reduced p-AKT protein expression.

CONCLUSIONThioridazine inhibits the proliferation and induces apoptosis of SW480 cells by up-regulating PDCD4 and inhibiting PI3K/Akt pathway.

Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Cycle Checkpoints ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Down-Regulation ; Humans ; RNA-Binding Proteins ; metabolism ; Signal Transduction ; drug effects ; Thioridazine ; pharmacology

BACKGROUNDThe success rate of antegrade approach for chronic total occlusions (CTO) recanalization has not dramatically increased, especially in complex CTO subset. The retrograde technique may hold great promise. This report aimed to describe our experience of retrograde recanalization for CTO, focusing on its safety and feasibility.

METHODSWe identified 42 patients who underwent revascularization in CTO with retrograde approach from July 2005 to November 2009 in our center.

RESULTSThree kinds of strategy were applied: retrograde as primary strategy (50.0%), retrograde immediately after antegrade failure (26.2%) and repeat procedure after previous antegrade failure (23.8%). Septal collaterals were more frequently used as the retrograde access route (92.9%). Overall success rate was 88.1%. In patients with successful retrograde wire crossing collateral channel to the distal cap of CTO, the success rate of recanalization was 94.1%. In patient with failure to cross the collaterals, the success rate was 62.5%. Eight different kinds of retrograde techniques were used: kissing wire technique (35.3%), wire trapped and reverse wire trapped technique (17.6%), back-end balloon and microcatheter reversal technique (14.7%), controlled antegrade and retrograde subintimal tracking (CART) technique (8.8%), reverse CART and modified reverse CART technique (8.8%), retrograde wire crossing technique (2.9%). There were 4 complications occurred without in-hospital major adverse cardiac events (MACE). In-hospital MACE was 7.7%. All of them were non-Q wave myocardial infarction. There were no cases of death or target vessel revascularization, either surgery or percutaneous.

CONCLUSIONSThe retrograde approach can be an effective tool for increasing the success rate of recanalization in the very complex CTO. To ensure the success and safety of the approach, careful case selection and device handling by experienced operators is essential.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Chronic Disease ; Coronary Angiography ; Coronary Occlusion ; therapy ; Female ; Humans ; Male ; Middle Aged ; Models, Theoretical ; Treatment Outcome

Objective To analyze the rates of occurrence,presentations and treatment of coronary intramural hematomas(IMH)after coronary artery stent implantation.Methods Retrospective analysis was carried out in non-chronic total occlusion patients who developed coronary intramural hematomas after coronary artery stent implantation between January 1,2011 to December 31,2016.Statistical analysis was made in the fields clinical data,coronary angiography features,treatment provided,and postoperative follow-up date of the patients.Results Among the 26 IMH patients,the male gender(15/26,57.7％)and existiing hypertension(17/26,65.4％)were more common risk factors for IMH after coronary artery stent implantation.Fourteen patients developed coronary dissection.The coronary intramural hematomas presented as new non-spasm and non-thrombus coronary stenosis.The coronary intramural hematomas were found to have involved the distal segment to the stents in 16 patients.Two patients received balloon dilation,five patients had stents implantation after balloon dilation,13 patients(50.0％)were treated with direct stent implantation and the other 6 patients did not have further intervention.The follow up period after hospital discharge was(2.39±1.68)years.No adverse cardiovascular event occurred.Five patients received follow-up angiography examination.Two patients and another one patient were found to have coronary intramural hematomas fully resolved at three months and one year with coronary angiographic follow up,respectively.Two patients had IMH on angiography at 1 year follow up.Conclusions Coronary intramural hematomas after coronary artery stent implantation often involved the distal segment to the stent in hypertensive patients presenting as new non-spasm and non-thrombus coronary stenosis.Patients at low risk of acute coronary occlusion could receive conservative treatment.Patients with extentsive length of intramural hematomas should consider stent implantation for treatment.

To investigate the antiviral efficacy of combination therapy with pegylated-interferon alpha (peg-IFNa)-2a and ribavirin (RBV) in hepatitis C patients with liver cirrhosis after splenectomy or partial splenic embolization. Forty-nine hepatitis C patients with liver cirrhosis who were unable to use antiviral therapy because of hypersplenism were recruited for study and treated with splenectomy or partial splenic embolization. Three months later, a regimen of antiviral combination therapy was initiated with peg-IFNa-2a (once-weekly subcutaneous injection: 135 μg or 180 μg) and RBV (daily oral: 800 to 1200 mg), and was maintained for 48 weeks. The patients were followed up at treatment weeks 1, 2, 4, 6, 8, and 12. Thereafter, follow-up was conducted every four weeks. The patients were observed until 24 weeks after treatment discontinuation. Follow-up testing included liver function, blood chemistry, renal function, and HCV RNA level. Any adverse reactions were recorded. Liver cirrhosis patients complicated by hypersplenism can be treated effectively with peg-IFNa-2a/RBV combination antiviral therapy after splenectomy or partial splenic embolization. The antiviral-induced sustained viral response rates was 65.00% in cirrhotic/hypersplenic hepatitis C patients receiving splenectomy and 58.62% in those receiving partial splenic embolization. Hypersplenism patients with hepatitis C-related cirrhosis achieved a good antiviral therapeutic effect with peg-IFNa-2a/RBV combination therapy following splenectomy or partial splenic embolization. This sequence of treatment may help to decrease incidences of chronic hepatitis C-induced liver failure and liver cancer in these patients.
Adult ; Aged ; Antiviral Agents ; therapeutic use ; Combined Modality Therapy ; Female ; Hepatitis C ; complications ; therapy ; Humans ; Interferon-alpha ; therapeutic use ; Liver Cirrhosis ; etiology ; therapy ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Postoperative Period ; Recombinant Proteins ; therapeutic use ; Ribavirin ; therapeutic use ; Splenectomy ; Treatment Outcome

OBJECTIVETo determine the prevalence of a common GJB2 mutation in a big Chinese population of deaf children and the features of its distribution in regions all over the nation and to provide epidemiology data and expertise for genetic testing of deafness in China.

METHODSThe DNA samples of NSHI patients and normal controls were collected from different typical areas of China. The method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with ApaI was used to determine the genotype of GJB2 235 site.

RESULTSTotally 16.3% of patients carried at least one 235 delC mutant allele. Among them, 7.8% was homozygous and 8.5% was heterozygous. The prevalence of GJB2 235delC mutation in China was evident, and the significant difference of 235delC mutation frequency was found in sub-population from different areas and different ethnic groups.

CONCLUSIONSBased upon the result of this screening as stated, Chinese NSHI patients appear to have 235delC frequency and the number of GJB2 related deafness was estimated to be huge. The testing of GJB2 235delC mutation would play an important role in genetic diagnosis and screening in China. As high as 15% of patients could be diagnosed as GJB2 caused deafness (bi-allelic mutation) only by means of this simple, fast and economic assay. In addition, patients were negative for 235delC mutation would be candidates for further mutational analysis of GJB2 or other deafness related genes.

Adolescent ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; China ; epidemiology ; Connexin 26 ; Connexins ; genetics ; Female ; Genotype ; Hearing Loss, Sensorineural ; epidemiology ; genetics ; Heterozygote ; Homozygote ; Humans ; Male ; Point Mutation ; Prevalence ; Young Adult

Objective:The goal of this study was to analyze the clinical significance of relationship between myocardial collateral and the levels of hypoxia‐inducible factor 1‐alpha (HIF‐1α) and vascular endothelial growth factor A (VEGF‐A) in patients with coronary chronic total occlusion lesion .Methods:89 patients with coronary chronic total occlusion lesion confirmed by clin‐ical data and coronary angiography were identified .The levels of HIF‐1αand VEGF‐A were measured by ELISA ,and the rela‐tive expression of VEGF‐A of peripheral blood mononuclear cell (PBMC) were measured by real‐time PCR .The results were statistically analyzed by the statistical programme for social sciences (SPSS version 18 .0) and software SAS JMP 9 .0 .Results:Compared to Rentrop 0‐1 grade group (18/38 ,47 .4% ) ,Rentrop 2 (11/31 ,35 .5% ) and Rentrop 3 (3/20 ,15 .0% ) grade group had fewer diabetes mellitus .Rentrop 2 [(6 .67 ± 1 .41) mmol/L] and Rentrop 3 [(5 .48 ± 1 .26) mmol/L] grade group had low‐er fasting blood glucose than Rentrop 0‐1 grade group [(7 .24 ± 1 .39) mmol/L] .Rentrop 2 (12/31 ,38 .7% ) and Rentrop 3 (3/20 ,15 .0% ) grade group had fewer clinical heart failure (NYHA Ⅱ ~ Ⅳ grade) than Rentrop 0‐1 grade group (20/38 , 52 .6% ) .Rentrop 2 [(85 .5 ± 27 .7) pg/mL ,(139 .5 ± 42 .1) pg/mL] and Rentrop 3 [(103 .3 ± 30 .2) pg/mL ,(162 .6 ± 43 .3) pg/mL] grade group had higher levels of HIF‐1αand VEGF‐A than Rentrop 0‐1 grade group [(42 .0 ± 16 .1) pg/mL ,(76 .5 ± 32 .2) pg/mL] .Rentrop 2 (1 .31 ± 0 .46) and Rentrop 3 (1 .38 ± 0 .44) grade group had higher level of relative expression of VEGF‐A in PBMC than Rentrop 0‐1 grade group (1 .00 ± 0 .28) .Conclusions:Chronic and consistent ischemia and hypoxia in‐duced the increase of expression of HIF‐1αand VEGF‐A is important for establishment of coronary collateral ,increasing blood supply and improving the heart function and prognosis .