Objective To investigate the effects of a miRNA family member, let-7e, and a combi-nation of miR-106b and miR-20a on the expression of cytokines by THP-1 cells with Luminex xMAP technol-ogy.Methods The efficiency of transfection was evaluated by immunofluorescence assay after transfecting THP-1 cells with micrONTM mimic negative control (Cy3) for 24 h, 36 h and 48 h.The three miRNA mim-ics (let-7e, miR-106b and miR-20a) were respectively used to transfect the THP-1 cells for 24 h, 36 h and 48 h and the expression of each miRNA was analyzed by qRT-PCR analysis for screening out the optimal transfection time.The transfected THP-1 cells were stimulated with1 mg/L of LPS for 1 h.The Luminex xMAP technology was used to detect the expression of IL-8, interferon-inducible protein-10 (IP-10), mono-cyte chemotactic protein 1 (MCP-1), IL-1α, IL-6, IL-10, TNF-α, IFN-αand IFN-βin the supernatants of cell culture.A statistical analysis was performed to analyze the data obtained by using SPSS16.0 software. Results More than 90% of the transfected THP-1 cells were labeled with red fluorescence.The optimal transfection times for let-7e mimic and miR-106b/miR-20a mimics were 48 h and 24 h, respectively.Com-pared with the corresponding negative control (NC), the expression of IL-8, IP-10 and MCP-1 by THP-1 cells were enhanced after the transfection with let-7e mimic, but were inhibited after the co-transfection with miR-106b and miR-20a mimics.Conclusion The expression of IL-8, IP-10 and MCP-1 were enhanced in let-7e transfected THP-1 cells, but were inhibited in miR-106b and miR-20a co-transfected THP-1 cells.

Objective To explore the pathological changes of lung, expression of the relevant inflammatory factors and oxidative stress markers of Sprague-Dawley (SD) rats undergoing autologous orthotopic liver transplantation (AOLT). Methods Thirty SD rats were randomized into sham group and AOLT group. The pathological changes of lung, expression of the relevant inflammatory mediators and oxidative stress markers were detected . Results ( 1 ) Compared with the sham group , the pathological scores of lung tissue in AOLT group increased significantly and reached its peak at 8 h after surgery. Then the pathological scores decreased to the level of sham within 24 h to 48 h after surgery; (2)The relative expression of inflammatory mediators including TNF-α, IL-1β, IL-6 and IL-8 increased significantly and reached its peak at 8 h after surgery in AOLT group. Then decreased to the level of sham group within 24 h to 48 h after surgery; (3)The change trends of MDA and H2O2 were similar to inflammatory mediators.The relative SOD expression decreased significantly and touched the nadir at 8h after surgery and then increased. Conclusion The pathological changes of lung expression, the relevant inflammatory mediators and oxidative stress markers of rats underwent AOLT were consistent.

Objective To comprehensively evaluate the clinical effect of cryoballoon ablation (CBA) and radiofrequency ablation (RFA) in treating paroxysmal atrial fibrillation (PAF).Methods Computer retrieval of PubMed,EMbase,the Cochrane Library,Web of Knowledge,China national knowledge infrastructure (CNKI),Chinese Biomedical Medical Literature (CBM),China Wan Fang,China VIP,and other database to collect the randomized control trials (RCT) related to RFA and CBA treatment for PAF.The retrieval time was from the establishment of database to December 2015.The data extraction and methodological quality of the included studies were assessed by two reviewers independently.And meta-analysis was conducted by using RevMan 5.2 software.Results A total of 6 research papers (636 patients in total) were included.The results of meta-analysis showed that the incidence of phrenic nerve paralysis in CBA group was significantly higher than that in RFA group (RR=9.26,95％CI:2.17-39.63,P=0.003).No statistically significant differences in the operation time (MD=10.07,95％CI:-9.10-30.52,P=0.29),fluoroscopy time (MD=-0.18,95％CI:-8.14-7.77,P=0.96),12-month success rate (RR=0.91,95％CI:0.72-1.14,P=0.40) and the incidences of atrial tachycardia,atrial flutter,atrioventricular reentrant tachycardia existed between CBA group and RFA group (RR=0.47,95％CI:0.11-2.02,P=0.31).Conclusion For the treatment of PAF,no obvious differences in the operation time,fluoroscopy time,12-month success rate,and the incidences of atrial tachycardia,atrial flutter,atrioventricular reentrant tachycardia exist between CBA and conventional RFA,but CBA can increase the incidence of phrenic nerve paralysis.

Objective To investigate the effect of Bcl-2 gene expression on the proliferation and apoptosis of human pancreatic cancer SW1990 cells.Methods Bcl-2 short guide RNA (Bcl-2-sgRNA) was designed and synthesized,and it was combined with CRISPR-Cas 9.After confirmation by gene sequencing,it was transfected into human pancreatic cancer cell line SW1990,then the cells with stable Bcl-2 gene knock-out were selected,and wild type SW1990 cells were used as control.The cell growth curve was determined by CCK-8 method.The number of clone formation was measured.Flow cytometry was used to measure cell cycle and apoptosis.Results Human pancreatic cancer cell line SW1990 with Bcl-2 gene knock-out was successful constructed.Compared with wild type SW1990 cells,the growth of SW1990 cells with Bcl-2 gene knock-out was inhibited,the number of clone formation was significantly decreased [(160.7 ± 10.0) vs (285.3 ± 14.2)],the proportion of G1 cells was significandy increased [(84.51 ± 0.97) ％ vs (57.49 ± 1.08) ％],the proportion of S phase cells significantly decreased [(12.82 ± 0.99) ％ vs (27.56 ± 1.65) ％],and apoptosis rate was remarkably increased [(12.67 ± 0.59) ％ vs (0.37 ± 0.35) ％],and the difference between the two groups was statistically significant (P ＜ 0.01).Conclusions Knock-out of Bcl-2 gene can inhibit the growth of human pancreatic cancer cell line SW1990,decrease the ability of clone formation,block the cell in G1 phase and greatly increase cell apoptosis rate.

Objective To explore the efficacy of sequential endoscopic variceal ligation plus endoscopic variceal sclerotheropy and the factors associated with the prognosis.Methods 106 cases with esophageal varices in control group was treated with drugs alone;study group had 113 cases, was given endoscopic therapy add drugs, The study group randomly divided into two groups, one was treated with endoscopic variceal ligation all the time (EVL group), another was treated with sequential endoscopic variceal ligation plus endoscopic variceal sclerotheropy (sequential group). After the treatment, the rate of removal of varicose veins, the rate of rebleeding, the number of total treatment,mortality and intra-operative complications and postoperative complications were compared. And compared rebleeding rate and mortality with the control group and study groups with different CTP and MELD, analyze the factors of prognosis, and evaluate their prognostic value.Results Rebleeding, rate in control group, EVL group and sequential group were 41.51%,10.53% and 10.64%,sequential group was significantly better than control group (P = 0.000); mortality in control group, EVL group and sequential group were 15.09%, 5.26% and 2.13%, sequential group was also significantly better than control group (P = 0.001); rate of recurrence in EVL group and sequential group within half a year were 73.68% and 44.68%, sequential group was significantly better than EVL group (P = 0.021). In all control group and EVL group and the sequential group, rebleeding rate and mortality of the liver function Child-Turcotte-Pugh (CTP) class C was significantly higher than that of calss A; In MELD model, AUC area under the ROC curve of rebleeding rate in control group and sequential group were 0.944 and 0.851, mortality of the two groups were 0.881 and 0.984, while the rate of recurrence in the EVL group and sequential group were respectively 0.914 and 0.765, the MELD score has the important value to the prediction of rebleeding and death.Conclusion The rebleeding rate and mortality in cirrhotic patients with esophageal varices treated with sequential endoscopic variceal ligation plus endoscopic variceal sclerotheropy were significantly decreased and the recurrence rate was lower than that of the patients with endoscopic variceal ligation all the time. Liver function Child-Turcotte-Pugh (CTP) score and the MELD score have important value in prediction of rebleeding and death, ligation and sclerosing sequential therapy can significantly reduced rebleeding and mortality in CTP class B and C, and improve the MELD threshold of rebleeding and death.

Hybrid operating room refers to clean operating rooms equipped with advanced MRI(magnetic resonance ima-ging), DSA(digital subtraction angiography) and CT(computer tomography), radiation therapy C-arm and navigation equip-ment.Currently, hospitals attach great importance to install and use DSA, so that the doctor can not only perform the conven-tional operation, but also directly explore the angiograpic and interventional therapy, which greatly improve the efficiency and the success rate of the surgery.After check out database, choosen about 40 papers of high quality.This paper introduces the concept of hybrid operating room, the development process, clinical application status and advantages in cardiovascular dis-ease, and prospects for its development in the future , and provides some suggestions to more easily build hybrid operating room.

Objective To assess the influence and early diagnosis treatment of bronchiectasis using severity score in patients with chronic obstructive pulmonary disease (COPD) combined with bronchiectasis. Methods Sixty patients with bronchiectasis in stable stage were selected from July 2016 to August 2017. The patients were divided into simple bronchiectasis group (32 cases) and bronchiectasis combined with COPD group (28 cases). The general clinical features, chest high-resolution CT (HRCT), pulmonary function, fractional exhaled nitric oxide (FENO) and bronchiectasis severity score between 2 groups were compared. The changes of the indexes 6 months after treatment with inhaled salmeterol fluticasone (50 μg/250 μg) in bronchiectasis combined with COPD group were observed. Results The bronchiectasis severity score and FENO in bronchiectasis combined with COPD group were significantly higher than those in simple bronchiectasis group: (4.82 ± 0.91) scores vs. (2.88 ± 0.83) scores and (39.04 ± 9.57) nmol/L vs. (28.66 ± 6.12) nmol/L, and there were statistical differences (P＜0.01). The forced expired volume in one second as a percentage of expected value (FEV1% Pred) and forced vital capacity as a percentage of expected value (FVC%Pred) in bronchiectasis combined with COPD group were significantly lower than those in simple bronchiectasis group: (61.36 ± 5.23)% vs. (71.28 ± 6.67)% and (61.57 ± 7.60)% vs. (72.84 ± 7.19)% , and there were statistical differences (P＜0.01). In bronchiectasis combined with COPD group, there were statistical differences in the bronchiectasis severity score, FEV1% Pred, FVC% Pred and FENO 6 months after treatment with inhaled salmeterol fluticasone and before treatment (P＜0.05). There was a negative correlation between FEV1% Pred and bronchiectasis severity score before and after treatment (r＝-0.802 and-0.618, P＜0.05); and there was a positive correlation between the FENO and bronchiectasis severity score (r ＝ 0.728 and 0.586, P＜0.05). Conclusions The pathogenetic condition of bronchiectasis combined with COPD is severe compared with simple bronchiectasis. Inhaled salmeterol fluticasone is effective in patients with bronchiectasis combined with COPD.

Mediator is a highly conserved large protein complex (25 proteins, >1000 kDa) and preeminently responsible for eukaryotic transcription, which contains a dissociable 'Cdk8 module'. Although increasing evidence demonstrates that Cdk8 module plays both positive and negative roles in transcription regulation, the detailed structure, and subunit organization, molecular mechanism how it regulates transcription remain elusive. Here we used single-particle electron microscopy to characterize the structure and subunit organization of the Cdk8 module and illuminated the substantial mobility of the Med13 subunit results in the structural flexibility. The Cdk8 module interaction with core Mediator is concurrent with active transcription in vivo. An interaction with the Cdk8 module induces core Mediator into very extended conformation in vitro, which is presumed to be an active functional state of Mediator. Taken together, our results illuminated the detailed architecture of Cdk8 module, and suggested the Cdk8 module could positively regulate transcription by modulating Mediator conformation.
Cyclin-Dependent Kinase 8 ; chemistry ; Humans ; Mediator Complex ; chemistry ; Models, Molecular ; Protein Conformation ; Protein Subunits ; chemistry ; Saccharomyces cerevisiae Proteins ; chemistry

OBJECTIVE: To carry out genetic testing for two families affected with cobalamin C (cblC) and establish a rapid method for the detection of a hotspot pathogenic variant c.609G>A of the MMACHC gene by using a PCR-high-resolution melting curve (PCR-HRM) method. METHODS: Genomic DNA was extracted from peripheral blood samples of the probands and their parents. Potential variants of the MMACHC gene was analyzed by Sanger sequencing. The c.609G>A variant of the MMACHC gene was screened among 100 healthy children with the PCR-HRM method. RESULTS: Sanger sequencing revealed that proband 1 carried compound heterozygous variants c.394C>T and c.609G>A of the MMACHC gene, while proband 2 carried compound heterozygous variants c.482G>A and c.609G>A of the same gene. PCR-HRM analysis of the two probands and the 100 healthy children were consistent with the Sanger sequencing. CONCLUSION c.609G>A is a hotspot pathogenic variant of the MMACHC gene. The diagnosis of cblC may be rapidly attained through detection by PCR-HRM.

Objective: To investigate the effects of anti-β₂ glycoprotein Ⅰ(β₂GPⅠ) antibody on atherosclerosis in ApoE deficient mice. Methods: A total of 24 male ApoE deficient mice of specific pathogen free level(six to eight-week old)were divided into normal control group, high fat diet group, high fat diet with anti-β₂GPⅠ group, high fat diet with homologous control antibody group (n=6 each group). During the feeding period, mice were weighed every 2 weeks and were intraperitoneally injected with anti-β₂GPⅠIgG (100 μg/per) and homologous control IgG (100 μg/per) according to grouping once a week. At the 16th week, the carotid arterial lipid deposition was observed by small animal magnetic resonance imaging, and blood samples were collected from internal vein of eyeball and the concentrations of TC, TG, HDL-C and LDL-C in plasma were measured after EDTA anticoagulant treatment. AI was calculated. The mice were then sacrificed and carotid arteries were removed, hematoxylin-eosin staining was used to observe the atherosclerotic lesions near the bifurcation of carotid artery and to calculate lesion size. Results: (1) The body weight of mice was significantly higher in the high fat diet group compared to other 3 groups(all P<0.05), which was similar among high fat diet+ anti-β₂GPⅠantibody group, high fat diet+ homologous control IgG group and normal diet control group (P>0.05). (2) After 16 weeks, plasma concentrations of TC and LDL-C in high fat diet group, high fat diet+ anti-β₂GPⅠantibody group and high fat diet+ homologous control IgG group were significantly higher than in normal diet group (all P<0.05), there was no significant difference among high fat diet groups. The level of HDL-C was significantly higher in high fat diet control group than in normal diet control group. The concentration of TG was similar among groups. However, the value of AI in high fat+ anti-β₂GPⅠ antibody group was significantly higher than in other groups (all P<0.05). (3) After 16 weeks, magnetic resonance imaging revealed that mice in high fat diet+ anti-β₂GPⅠ antibody group had more obviously lipid deposition in the carotid arteries, it was significantly higher than that in the other groups, and the cross sections of carotid arteries stained with HE also demonstrated obviously carotid lumen stenosis and the percentage of carotid plaque area to carotid artery was (37.545±1.351)% in the high fat diet+ anti-β₂GPⅠ antibody group, it was significantly higher than normal diet group ((1.235±0.460)%), high fat diet control group((11.635±2.751)%) and high fat diet+ homologous control IgG group ((11.815±2.623)%), all P<0.01. In high fat diet+ anti-β₂GPⅠ antibody group, the area of carotid plaque was (3.121±0.124)×10⁴ μm^2, it was also significantly higher than normal diet group ((0.094±0.015)×10⁴ μm²), high fat diet control group ((1.309±0.147)×10⁴ μm²) and high fat diet+ homologous control IgG group ((1.027±0.228)×10⁴μm²), all P<0.01. Conclusion Anti-β₂GPⅠ antibody can promote atherosclerotic plaque formation in high fat diet fed ApoE deficient mice.