The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2％to 99.1％,with the highest homology of 99.1％observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.

The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2％to 99.1％,with the highest homology of 99.1％observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.

Objective:To investigate the effect of miR-199a on interstitial transdifferentiation (EMT) of human renal tubular epithelial cells stimulated by high glucose.Methods:Human renal tubular epithelial cells were cultured in vitro and divided into low glycemic group (LG), high osmotic group (HM) and high glycemic group (HG) according to different glucose concentrations. They were divided into miR-199a inhibition group (miR-199ai group), negative control group (miR-INC group) and liposome group (Mock group) according to whether they were transfected with miR-199a inhibitors (miR199ai). They were divided into si-hypoxia-inducible factor-1α (si-HIF-1α) group, miR-199ai group, miR-199ai+ si-HIF-1α group and negative control group (NC group) according to whether they converted to si-HIF-1α and miR-199ai. The expressions of miR-199a, HIF-1α, fibronectin (FN) and α-smooth muscle actin (α-SMA) in different groups of cells were detected by real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) and Western blot. Dual luciferase target assay was used to verify the targeting relationship between miR-199a and HIF-1α gene.Results:The mRNA expression level of miR-199a in the HG group was significantly lower than that in the LG group and the HM group (all P<0.01), and the mRNA and protein expression levels of HIF-1α, FN and α-SMA in Hg group were significantly higher than those in LG group and HM group (all P<0.01). The mRNA and protein expression levels of HIF-1α, FN and α-SMA in the miR-199ai group were significantly higher than those in the miR-iNC group and the Mock group (all P<0.01). Target gene prediction by TargetScan and miRanda showed that miR-199a could target the 3′-untranslated region(3′-UTR) that binds HIF-1α. Dual luciferase reported results: Inhibition of miR-199a resulted in a significant increase in luciferase activity ( P<0.01), but there was no significant change in luciferase activity after 3 ′-UTR mutation of HIF-1α ( P>0.05). After transfection with si-HIF-1α, the mRNA and protein expression levels of HIF-1α, FN and α-SMA in si-HIF-1α group were significantly lower than those in NC group (all P<0.05). After transfection with miR-199ai, the mRNA and protein expression levels of HIF-1α, FN and α-SMA in miR-199ai group were significantly higher than those in NC group (all P<0.05). However, when co-transfected with si-HIF-1α and miR-199ai, the expression levels of mRNA and protein in miR-199AI group were significantly higher than those in NC group (all P<0.05). The mRNA and protein expression changes of HIF-1α, FN and α-SMA induced by transfection of si-HIF-1α or miR-199ai alone can be recovered. Conclusions:miR-199a can alleviate hyperglycemia-induced EMT in human renal tubular epithelial cells through targeted regulation of HIF-1α.

Objective:To explore the relationship between the subcomponents of executive function and situation awareness.Methods:A total of 73 cadets of our school were selected to complete a flight simulation mission. The situation awareness rating technique (SART) scores, situation awareness global assessment technique (SAGAT) scores and flight performance were recorded as dependent variables. Spatial n-back task, Go/Nogo task, Stroop task and digital shifting task were used to measure the subcomponents of executive functions and perform independent multiple linear regression analysis as predictive variables.Results:Pearson correlation analysis showed that updating score was significantly and positively correlated with response inhibition, SART, SAGAT and flight performance ( r=0.235, 0.285, 0.298, 0.264, P=0.049, 0.016, 0.012, 0.025), response inhibition score was significantly and positively correlated with SAGAT and flight performance ( r=0.349, 0.243, P=0.003, 0.041), interference control score was only negatively correlated with cognitive shifting and flight performance ( r=-0.350, -0.235, P=0.003, 0.045), cognitive shifting score was significantly and positively correlated with SART, SAGAT and flight performance ( r=0.249, 0.275, 0.364, P=0.039, 0.019, 0.002). Further regression analysis indicated that updating score has a significant predictive effect on SART ( t=2.201, P=0.031), updating and response inhibition scores have significant predictive effects on SAGAT ( t=2.270, 2.546, P=0.026, 0.013), and updating and cognitive shifting scores have significant predictive effects on flight performance ( t=2.508, 2.553, P=0.015, 0.013). Conclusions:The subcomponents of executive function are significantly with the level of situation awareness and have predictive significance.

Objective:To explore the relationship between the subcomponents of executive function and situation awareness.Methods:A total of 73 cadets of our school were selected to complete a flight simulation mission. The situation awareness rating technique (SART) scores, situation awareness global assessment technique (SAGAT) scores and flight performance were recorded as dependent variables. Spatial n-back task, Go/Nogo task, Stroop task and digital shifting task were used to measure the subcomponents of executive functions and perform independent multiple linear regression analysis as predictive variables.Results:Pearson correlation analysis showed that updating score was significantly and positively correlated with response inhibition, SART, SAGAT and flight performance ( r=0.235, 0.285, 0.298, 0.264, P=0.049, 0.016, 0.012, 0.025), response inhibition score was significantly and positively correlated with SAGAT and flight performance ( r=0.349, 0.243, P=0.003, 0.041), interference control score was only negatively correlated with cognitive shifting and flight performance ( r=-0.350, -0.235, P=0.003, 0.045), cognitive shifting score was significantly and positively correlated with SART, SAGAT and flight performance ( r=0.249, 0.275, 0.364, P=0.039, 0.019, 0.002). Further regression analysis indicated that updating score has a significant predictive effect on SART ( t=2.201, P=0.031), updating and response inhibition scores have significant predictive effects on SAGAT ( t=2.270, 2.546, P=0.026, 0.013), and updating and cognitive shifting scores have significant predictive effects on flight performance ( t=2.508, 2.553, P=0.015, 0.013). Conclusions:The subcomponents of executive function are significantly with the level of situation awareness and have predictive significance.

Objective:To explore the clinical effect of azacitidine combined with CAG regimen on the treatment of relapsed/refractory acute myeloid leukemia (AML).Methods:The data of 50 patients with relapsed/refractory AML (non-acute promyelocytic leukemia) in Jining No. 1 People's Hospital from September 2018 to September 2019 was retrospectively analyzed, and the patients were divided into the control group and the test group according to the different treatment drugs. The control group (28 cases) was treated with CAG regimen alone, and the test group (22 cases) was treated with azacitidine combined with CAG regimen. The total effective rate and adverse reactions of the two groups were observed after 1 course of treatment.Results:After one course of treatment, the total clinical effective rate in the test group was 86% (19/22), and the rate in the control group was 71% (20/28), there was a statistically significant difference between the two groups (χ 2 = 5.273, P < 0.05). There were no significant differences in the incidence of adverse reactions such as fever, pulmonary infection, vomiting, and thrombocytopenia between the two groups (all P > 0.05). Conclusion:Azacitidine combined with CAG regimen in the treatment of relapsed/refractory AML can improve the clinical efficacy without increasing the adverse reactions.

Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Interferon-alpha ; Leukemia, Promyelocytic, Acute ; Tretinoin

Objective To investigate the effect of umbilical cord blood dendritic cells(DCs)induced by gastric cancer antigen combined with cytokine induced killer(CIK)cells in gastric cancer cell lines SGC-7901 in vitro. Methods Mononuclear cells from umbilical cord blood were used to create DCs and CIKs. The cell surface antigen expression of the mature DCs such as CD83,CD86,CD11c and the cell surface antigen of CIKs such as CD3,CD56,CD4,CD8,CD16 were detected using flow cytometry. Sensitized DCs-CIKs,DCs-CIKs,CIKs as effective cells,and SGC-7901 as target cells,the killing activities of these effective cells were tested with LDH release,which the number ratio of cells between effective cells and SGC-7901 cells were 10 :1,20 : 1,40 : 1,respectively. Results The cell surface antigen expressions of the mature DCs,such as CD83 + CD86 + ,CD11c + CD83 + ,CD86 + CD11c + were(75. 4 ± 2. 1)% ,(79. 3 ± 1. 4)% ,(80. 2 ± 2. 6)% , respectively. The mature sensitive-DCs surface antigen expressions,such as CD83 + CD86 + ,CD11c + CD83 + , CD86 + CD11c + ,were(77. 7 ± 1. 5)% ,(82. 6 ± 1. 9)% ,(76. 9 ± 2. 6)% ,respectively. There was no sta-tistical significance about the surface antigen expression between DCs and sensitive-DCs(t = 1. 526,P ﹥ 0. 05;t = 0. 958,P ﹥ 0. 05;t = 1. 049,P ﹥ 0. 05). The CIKs surface antigen expressions,such as CD4 + ,CD8 + , CD3 + CD56 + CD16 + ,were(22. 8 ± 1. 3)% ,(77. 3 ± 1. 8)% ,(24. 5 ± 2. 1)% ,respectively. The results suggested that the killing effect of the three kinds of combination cells on gastric cancer cells was different. The number ratio of cells between sensitive-DCs and SGC-7901 cells were 10 : 1,20 : 1,40 : 1,which the killing activities of sensitive-DCs-CIKs against SGC-7901 were(37. 68 ± 1. 49)% ,(41. 67 ± 0. 90)% ,(42. 71 ± 0. 98)% ,respectively. The killing activity of sensitive-DCs-CIKs was the highest when the ratio of cells between sensitive-DCs and SGC-7901 cells were 40 : 1. The killing activities of DC-CIKs were(36. 77 ± 0. 46)% ,(38. 94 ± 0. 95)% ,(41. 15 ± 0. 89)% ,respectively. The killing activities of CIKs were(34. 74 ± 1. 01)% ,(37. 76 ± 0. 43)% ,(39. 65 ± 0. 79)% ,respectively. There were statistically significant differences among the three groups(F = 5. 92,P ﹤ 0. 05;F = 19. 13,P ﹤ 0. 05;F = 8. 88,P ﹤ 0. 05). Conclusion The tumor killing activity of CIK is enhanced obviously by umbilical cord blood DCs which is sensitized by gastric cancer tumor antigen. There is the highest killing activity when the number ratio of cells between sensitive-DC-CIK and SGC-7901 cells is 40 : 1.

Inrecentyearsmanytargetedagentstreatinggastriccancerhavebeenevaluatedinclinical studies.Trastuzumab,an anti-HER-2 monoclonal antibody,has shown activity against HER-2-positive gastric cancer and become the first targeted agent approved in gastric cancer.Drugs targeting epidermal growth factor receptor,including monoclonal antibody and tyrosine kinase inhibitor,bring survival benefit to patients with gastric cancer.Addi tionally,vascular endothelial growth factor inhibitors are also under investigation.Other targeted agents are in preclinical or early clinical development,such as m-TOR inhibitors and c-MET inhibi-tors.

Objective To evaluate the therapeutic efficacy of ureteroscopic pneumatic lithotripsy for treatment of complicated ureteral calculi. Methods Clinical data of 56 cases with complicated ureteral calculi who were treated with ureteroscopic pneumatic lithotripsy was reviewed. Among them 15 cases were proximal ureteral calculi,41 cases were middle and distal ureteral calculi. Treatment by extracorporeal shockwave lithotripsy (ESWL) failed in 25 cases,8 cases complicated with fever. Results In 41 cases with middle and distal ureteral calculi, successful lithotripsy was achieved after the first treatment session,the success rate was 100.0% (41/41). In 15 cases with proximal ureteral calculi ,successful lithotripsy was achieved in 10 cases after the first treatment session,the success rate was 66.7%(l0/15). Calculi migrated to kidney in 3 cases who treated by ESWL for 1-2 times. The failure was due to ureteral kink in 1 case and due to ureteral perforation that required surgical repair in another case. Forty-nine cases were calculi-free after 4 weeks,the calculi-free rate was 87.5%(49/56). Emergency operation was performed in 8 cases complicated with fever. Successful lithotripsy was achieved after the first treatment session in 7 cases. In one patient,calculi migration occurred. Within all 8 cases obstruction was released and fever was allayed after operation.The complication rate was 1.8% (1/56). Conclusion Ureteroscopic pneumatic lithotripsy for treatment of complicated ureteral calculi is a safe and effective method.