Objective To explore the mechanism by which zinc-regulated transporters, iron-regulated transporter-likeprotein 4 (ZIP4) regulates glycolysis and its impact on tumor progression in cholangiocarcinoma (CCA), providing a theoretical basis for targeted therapy of CCA. Methods ZIP4 expression in CCA was analyzed using the GEPIA database. Immuno-histochemistry (IHC) was used to detect ZIP4 expression in 20 paired CCA and adjacent non-tumor tissues. Stable ZIP4-overexpressing CCA cell lines (ZIP4-OE) were established. Gene set enrichment analysis was used to screen differentially expressed genes and pathways in ZIP-OE CCA cells. ZIP4, N-myc proto-oncogene protein (MYCN), and histone-lysine N-methyltransferase 2E (KMT2E) were knocked down using small interfering RNAs (siRNAs). The expression of glycolysis-related gene (glucose transporter 1 [Glut1], hexokinase 2 [HK2], and lactate dehydrogenase A [LDHA]) was measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Glycolytic activity was assessed by measuring the extracellular acidification rate (ECAR). Cell proliferation was evaluated using colony formation assays, and cell migration was assessed using Transwell assays. A xenograft mouse model was constructed to examine CCA tumor growth. Protein levels of ZIP4, KMT2E, H3K4me3 (tri-methylation of lysine 4 on histone H3), and MYCN were detected by Western blotting. Results GEPIA database analysis and IHC results confirmed significantly higher ZIP4 expression levels in CCA tissues compared to adjacent non-tumor tissues (P＜0.05). Compared to the control group, the ZIP4-OE group exhibited a significantly increased ECAR, along with significantly enhanced proliferation and migration abilities (P＜0.01). Conversely, knockdown of ZIP4 suppressed CCA cells proliferation and migration. GEPIA analysis indicated that ZIP4 upregulates the transcription of oncogene MYCN, as well as glycolysis-related genes. Knockdown of MYCN abolished the ZIP4 overexpression-induced upregulation of Glut1, HK2, and LDHA gene transcription, reduced glycolysis, and significantly inhibited CCA cell proliferation and migration (P＜0.05). Mechanistic studies demonstrated that ZIP4 increases H3K4me3 level via KMT2E, leading to MYCN transcription. Knockdown of KMT2E in CCA cells suppressed the ZIP4 overexpression-induced enhancement in H3K4me3 modification, resulting in MYCN downregulation and significantly reduced CCA cells proliferation and migration (P＜0.05). Conclusions ZIP4 upregulates H3K4me3 modification through KMT2E, which recruits transcription factors to activate the transcription of MYCN. This subsequently enhances cellular glycolysis and promotes the proliferation and migration of CCA cells.

Objective:To analyze the biological characteristics, phylogenic features and clinical significance of SQ219 and SQ220 isolated from clinical sputum and midstream urine specimens.Methods:The culture and biochemical characteristics of the two strains were observed. VITEK2 System, drug sensitivity testing and MALDI-TOF mass spectrometry were used for bacterial identification. Phylogenetic analysis based on 16S rRNA and core genome was performed. The average nucleotide identity (ANI) based on whole genome sequences was calculated.Results:SQ219 and SQ220 were Gram-stain-negative, aerobic, catalase- and oxidase-positive, and non-motile bacteria. Their optimum growth was observed in NaCl-free medium at 30℃ and pH7. Flexirubin-type pigments were produced by SQ220 on Colombia blood agar, but not by SQ219. Both SQ219 and SQ220 were resistant to aztreonam, amikacin, tobramycin and colistin, which was consistent with the drug resistance phenotype of genus Chryseobacterium. The genome sequences of SQ219 and SQ220 were 5.08 Mb and 4.80 Mb in length, and the G+ C contents were 36.72% and 36.36%, respectively. Both strains carried β-lactam resistance gene ( blaCGA). 16S rRNA phylogenetic analysis showed that SQ219 and SQ220 were closely related to Chryseobacterium gambrini DSM18014 T with the similarities of 98.93% and 98.36%, respectively. Core genome phylogenetic analysis revealed that SQ219 and SQ220 were highly homologous to Chryseobacterium gambrini DSM18014 T. However, the ANI values between the two strains and Chryseobacterium gambrini DSM18014 T were 92.49% and 93.27%, respectively, below the threshold for prokaryotic species identification. Conclusions:Based on the phenotypic and phylogenetic data, SQ219 and SQ220 represent a novel species of the genus Chryseobacterium. This study would help promote the understanding of the evolution of Chrysobacterium and provide reference for the identification of new species of Chrysobacterium.

Objective: To investigate the oxidative stress injury of nano zinc oxide nanoparticles(ZnO NPs) on human myocardial cells (AC16) ，and to analyze the mechanism of ZnO NPs from the transcriptome level． Methods: Dynamic light scattering (DLS) was used to characterize and detect ZnO NPs．After AC16 cells were exposed to ZnO NPs at different doses and at different times，the cell survival rate was determined by CCK-8 method．AC16 cells were divided into control group，ZnO NPs (50，100，200 μmol /L) ，after 6 h treatment，the mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were measured．AC16 cells were divided into control group，50 μmol /L ZnO NPs group and 200 μmol /L ZnO NPs group．After 6 h exposure，total RNA was extracted by TRIzol for transcriptome analysis ，and the differentially expressed genes were enriched by gene body ( GO) ，Kyoto Encyclopedia of Genes and Genomes (KEGG) . Results : The results of DLS showed that the hydrodynamic diameter was ( 192. 2 ± 1. 63 ) nm and the Zeta potential was ( －23. 26 ± 1. 05 ) mV． CCK-8 results showed that the survival rate of AC16 cells decreased with the increase of dose and time of exposure to ZnO NPs. Fluorescence quantification showed that with the increase of ZnO NPs exposure dose，MMP significantly decreased at 100 μmol /L ZnO NPs(P＜0. 05) ，and ROS significantly increased at 50 μmol /L ZnO NPs(P＜0. 05) ．Using the multifunctional microplate reader，it was observed that MMP and ROS were statistically significant at 100 and 50 μmol /L ZnO NPs，respectively，showing a decrease in MMP and an increase in ROS．Transcriptome analysis showed that 1 071 genes were enriched in the 50 μmol /L ZnO NPs group compared with the control group，including 561 up-regulated genes and 510 down-regulated genes．Compared with the control group，7 164 genes were enriched in 200 μmol /L ZnO NPs group，including 4 098 up-regulated genes and 3 066 down-regulated genes．GO and KEGG analysis showed that the differential genes were mainly concentrated in ROS，antioxidant activity，mitochondrial cytochrome C release，apoptosis and other signaling pathways． Conclusion ZnO NPs can decrease the survival rate of AC16 cells and induce mitochondrial damage and oxidative stress，among which ROS-mediated oxi- dative stress and mitochondrial function changes are important toxic mechanisms of ZnO NPs induced AC16 cytotoxicity.

Objective:To analyze the application of thinking visual guidance teaching model in anesthesia clinical practice teaching.Methods:Taking the implementation time (March 2019) of thinking visual guidance teaching model in our hospital as the limit, 56 interns who came to our hospital for anesthesia practice before the implementation were included as control group, and 61 interns enrolled after the implementation (from March 2019 to March 2020) were included in study group. The mastery status of professional skills (duration of anesthesia operation, success rate of one-time anesthesia puncture and success rate of one - time tracheal intubation), professional knowledge assessment results and professional attitude changes were compared between the two groups at 2 months and 6 months of training. Self-evaluation after 6 months of training was compared between the two groups. SPSS 18.0 was used for t test and chi-square test. Results:After 6 months of training, the mastery status of professional skills (duration of anesthesia operation, success rate of one-time anesthesia puncture, and success rate of one-time tracheal intubation) of the two groups of interns were significantly improved compared with those after 2 months of training, and the above indicators of study group were significantly better than those of control group (all P<0.05). After 6 months of training, the scores of assessment results of professional knowledge (basic knowledge, understanding and memory, case analysis, emergency handling) and professional attitude (behavior, language, initiative and adaptability) in the two groups of interns were significantly higher than those after 2 months of training, and the above indicators scores of study group were significantly higher than those of control group (all P<0.05). The scores of self-evaluation (learning efficiency, learning atmosphere, learning ability, self-confidence and satisfaction) of study group at 6 months of training were significantly higher than those of control group at the same period (all P<0.05). Conclusion:Thinking visual guidance teaching model can effectively improve the professional knowledge and operation skills of anesthesia clinical interns, and it has a good application effect.

Rheumatoid arthritis (RA) is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes (FLS). The up-regulated cellular membrane expression of G protein coupled receptor kinase 2 (GRK2) of FLS plays a critical role in RA progression, the increase of GRK2 translocation activity promotes dysfunctional prostaglandin E4 receptor (EP4) signaling and FLS abnormal proliferation. Recently, although our group found that paeoniflorin-6'-

Objective To study the use of preoperative indocyanine green retention test at 15 minutes (ICG R15) in the prediction of liver functional reserve in patients with hilar cholangiocarcinoma (HCCA).Methods The clinical data of 62 patients with HCCA treated in our department from March 2016 to March 2018 was reviewed.The relationship between preoperative ICG R15 and postoperative hepatic insufficiency was analyzed.The relationship between preoperative ICG R15 and Child-Pugh scoring was also studied.Univariate analysis was used to evaluate the risk factors of postoperative liver dysfunction.Logistic regression was used to assess the independent risk factors of postoperative liver dysfunction.The regression equation between independent risk factors and postoperative liver dysfunction was established.Results Among the 62 patients,ICG R15 was less than 10.0％ in 26 patients,between 10.0％ and 19.0％ in 17 patients,between 20.0％ and 29.0％ in 9 patients,between 30.0％ and 39.0％ in 5 patients,and over 40.0％ in 5 patients.There were 29 patients with a Child-Pugh A grading and 33 patients with a Child-Pugh B grading in the preoperative evaluation of liver function.The Wilcoxon W rank sum test was used to compare the preoperative ICG R15 in patients with Child-Pugh grading A and B separately.The ICG R15 in Child-Pugh grading A patients was significantly lower than those in Child-Pugh B grading patients (P ＜0.05).There were no significant differences in age,gender,history of previous liver diseases,duration of operation,and intraoperative blood loss (P ＞ 0.05) between the normal liver function group and the liver dysfunction group.However,there was a significant difference in the preoperative ICG R15 and preoperative bilirubin levels (P ＜ 0.05) between the two groups.The preoperative ICG R15 and preoperative bilirubin levels were significant risk factors of postoperative hepatic insufficiency.Regression analysis suggested that preoperative ICG R15 level was an independent risk factor of postoperative hepatic insufficiency (P ＜ 0.05).A regression equation:logit(P) =0.185 × preoperative ICG R15-3.152 could be constructed.Conclusions ICG R15 is an ideal clinical indicator for evaluation of preoperative liver functional reserve in patients with HCCA.It predicted the recovery of postoperative liver function.

Objective:To identify the risk factors of anxiety disorders among patients undergoing coronary angiography (CAG) and to determine whether the decision of revascularization affect anxiety level following coronary angiography.
Methods:A total of 379 patients undergoing CAG in Fuwai Hospital from Dec. 2012 to Dec. 2013 were invited to participate this study. A data-collecting form, which included questions about demographic features, health history, Type A Behavior Questionnaire (TABQ) and Self Rating Anxiety Scale (SAS), was completed by the participants on the day before and the day after CAG.
Result:Among these patients, SAS score of both before and the day after CAG were higher than Chinese normative SAS score. Female patients had a higher SAS score level than male patients (40.57±9.53 vs 38.26±9.61, P<0.05) before CAG. Patients with these factors of female, over 50 years old, duration of coronary artery disease over 1 year, lower education level had a higher SAS score after CAG. SAS score declined signiifcantly after CAG except those scheduled to receive CABG. Multivariable linear analysis found the D-value between SAS scores before and after CAG was negative correlated with degree of education and positive correlated with the duration of coronary artery disease.
Conclusion:The anxiety level decreased after CAG, except those who need coronary revascularization surgery.

Purpose To assess the clinical application of non-contrast-enhanced MR angiography (NCE-MRA) using flow sensitive dephasing (FSD) prepared steady-state free precession (SSFP) for displaying hand arteries of patients with rheumatoid arthritis. Materials and Methods Twenty-two patients with rheumatoid arthritis were recruited in this study. All the patients undertook hand NCE-MRA and three-dimensional dynamic CE-MRA on a 1.5T MR scanner. The informed consent was obtained from each subject. Image quality was assessed independently by two experienced radiologists at three arterial segments (wrist arteries, palm arteries, andfinger arteries) with a four-point scale. Signal to noise ratio (SNR), contrast to noise ratio (CNR), and vessel sharpness were evaluated by a magnetic resonance physicist. The results and image quality were statistically compared between the two MRA techniques.Results Twenty-two patients of 24 hands successfully underwent NCE-MRA and CE-MRA scan. Among 72 vascular segments, 69 segments of NCE-MRA were diagnostic, which was higher than that of CE-MRA (96% vs 83%,P<0.05). Otherwise, the image quality, SNR, CNR and vessel sharpness of NCE-MRA were all superior to those of CE-MRA (P<0.05).Conclusion NCE-MRA using FSD-prepared SSFP allows clear depiction of the hand arterial tree, and the image quality is superior to that of dynamic CE-MRA. It is a potential tool for evaluating the disease of hand arteries.

Objective To explore the related factors on the mental health and the correlation between mental health status and social support and coping style in patients with occupational diseases in order to provide the scientific basis for making appropriate countermeasures to system intervention .Methods Eighty-seven patients with occupation diseases in our hospital were surveyed by the symptom checklist 90 ( SCL-90 ) , social support rating scale (SSRS), coping style questionnaire (CSQ).Results The mental health status of patients with occupational diseases was poor , and the scores of all factors on SCL-90 in patients were higher than the national norm, and the difference was statistically significant (P<0.01).The scores of all factors on social support in patients were significantly lower than the national norm , and the difference was statistically significant (P<0.01).The scores of total SSRS, the factors on objective support and support utilization had negative correlation with the scores of total SCL-90 and the part factors in patients (P<0.05).The scores of the part factors on SCL-90 had positive or negative correlation with the scores of the part factors on CSQ (P<0.05), but had not relation with the mature or immature coping style in patients .The scores of problem solving and mature coping style had positive correlation with the scores of subjective support and the total SSRS in patients ( P<0.05).Conclusions The patients with occupation diseases should be given more social support and care in clinical practice , and we should help them adjust mental status and improve the coping style and maintenance of physical and mental health .

BACKGROUND: Recent researches demonstrate that transplantation of bone marrow stem cells in the area of myocardial infarction can directionally differentiate into myocardial cells having normal physiological function and can promote newborn vascularization so as to repair infarction myocardium and improve injured cardiac function.OBJECTIVE: To observe short-term clinical effect of autologous bone marrow stem cell transplantation (ABMSCT) in percutaneous coronary artery on the treatment of acute myocardial infarction.DESIGN: Self-control study.SETTING: Department of Cardiology, Xiangtan Central Hospital.PARTICIPANTS: A total of 27 patients with acute myocardial infarction, including 16 males and 11 females, were selected from Department of Cardiology, Xiangtan Central Hospital from June 2004 to December 2006. Their ages coronary artery was finished after onset emergently; in addition, blood flow of infraction related vessels recovered to grade TIMI3. All patients provided the confirmed consent.METHODS: Operative procedure: All patients were performed with emergently interventional therapy of coronary artery after onset of acute myocardial infarction. One week later, percutaneous cavity tube technique was used to establish infarction related arterial pathway, and guiding filament was used to send micro-perfusion tube into stents. And then,separated bone marrow stem cell suspension was poured through central cavity of micro-tube into the distal end of infarction vessels. Operative evaluation: Dynamic electrocardiogram was evaluated for 24 hours before and after transplantation; in addition, left ventricular ejection fraction and myocardial perfusion defect scores were detected before and at 6 and 12 months after transplantation; otherwise, recovery state and complication were observed in follow up at 6 and 12 months after operation.defect scores: At 6 and 12 months after operation, left ventricular ejection fraction was higher than that before transplantation, and there was significant difference before and after transplantation (P＜0.05). While, myocardial perfusion defect scores were lower than those before transplantation, and there was significant difference before and arrhythmia were not found out, cardiac arrhythmia was not increased, and cardiac arrhythmia combining with malignancy not have any complications and in-stent constriction after operation.