Objective To observe the effect of tanshinone IIA on the myocardial calmodulin (CaM) and calmodulin dependent protein kinase II (CaMK) mRNA expression and heart function of acute myocardial infarction rats. Methods A total of 100 rats were randomly divided into the sham operation group, the model group, and the tanshinone ⅡA group. The model of acute myocardial infarction was established by the ligation of left anterior descending coronary artery ligation with 45 min, and reperfusion with 45 min, and the above process was repeated 3 times. Tanshinone IIA group was treated with intravenous injection of 10 mg/kg sodium tanshinone ⅡA, while the sham operation group and the model group with 10 mg/kg saline, and all treatment last 7 days. The mRNA expressions of CaM and CaMKII were determined. TTC staining was used to observe areas of myocardial infarction. The left ventricular maximum rise/fall rate (±LVdp/dtmax), myocardial contraction tension, heart rate and heart function index were detected pre-and post-treatment by the Langendorff device perfusion of isolated heart. Results Compared with the model group, the mRNA expression of and CaM mRNA (1.29 ± 0.19 vs. 2.31 ± 0.21) and CaMK II (1.10 ± 0.07 vs. 2.13 ± 0.18) in the tanshinone IIA group were significantly lower (P<0.05), and the areas of myocardial infarction (25.12%± 0.43%vs. 35.15%± 0.64%) significantly decreased (P<0.05), and the IT (2.03 ± 0.14 g vs. 1.06 ± 0.12 g), the ±LV dp/dtmax (4 701.2 ± 135.3 mmHg/s vs. 3 214.7 ± 110.2 mmHg/s, 2 518.7 ± 65.4 mmHg/s vs.1 960.3 ± 62.5 mmHg/s) significantly increased (P<0.05). Conclusion Tanshinone IIA may protect myocardial ischemia by regulating the mRNA expression of CaM and CaMKⅡ.

Objective To investigate the diagnosis value of immunophenotype and karyotypes in newly diagnosed chronic lymphocytic leukemia (CLL).Methods To retrospect the flow cytometry (FCM) immunophenotype and karyotypes characteristics in newly diagnosed 70 CLL cases.Results In all cases,the positive rates of CD19,CD20,CD5,CD23,CD22 were 100 ％,88.5 ％ (54/61),77.1％ (54/70),67.6 ％ (23/34)and 51.9 ％ (27/52),respectively.And 6 were misdiagnosed,2 was CD+5CD+19(+),but CD20,CD22 were strongly positive,final diagnosed as mantle cell lymphoma (MCL) by FISH t(11;14) examination and CyclinDl; CD+5CD+19(-) CLL were 16 cases (22.9 ％),but 4 were misdiagnosed,the misdiagnosis rate was 25 ％,significantly higher than that of CD+5CD+19(-) CLL (P =0.030).59 cases were examined by conventional cytogcnetic (CC),and 13 cases were with abnormal karyotypes,positive rate was 22.0 ％,with complex karyotypes in 5 cases (8.5 ％); 10 cases combined with FISH abnormalities karyotype examination rate was 60 ％.Conclusion Typical CLL immunophenotypic characteristics were CD5,CD19,CD23 co-expression,and CD－5 CLL with higher misdiagnosis rate,combined with CD20 (,) CD22 expression and karyotype analysis helps to CLL and other B lymphoid proliferative diseases (B-LPD) identification.Conventional cytogenetic detection combined with FISH scan can improve the recognition ability of abnormal chromosome.

Objective To investigate the factors affecting the susceptibility of tigecycline and assess the testing methods.Methods The 116 isolates of Acinetobacter baumanaii and Klebsiella pneumoniae were collected in 13 hospitals from January to December,2010,to evaluate the effects on the tigecycline susceptibility of the overnight medium,medium brand and lot number,respectively.The 56 isolates of Acinetobacter baumannii and the 47 isolates of Klebsiella pneumoniae were selected randomly according to the MIC distribution proportion in 2010 and 2012.The broth microdilution was taken as the reference method to evaluate the effects of the agar dilution,disk diffusion,MIC Test Strip (MTS) and Vitek2 (GN16) on the susceptibility of tigecycline.Results The essential agreement (EA) of Acinetobacter baumannii and Klebsiella pneumoniae is 89.7％ (52/58)and 87.9％ (51/58) using overnight medium and fresh medium respectively.Both EA and categorical agreement (CA) of the different brands (BBL and Oxoid) and lot numbers are 100％ using agar dilution.According to the FDA break point criteria,the CA/EA is 77.7％ (80/103)/99.0％ (102/103),87.4％ (90/103)/98.1％ (101/103),64.1％ (66/103)/76.7％ (79/103) using agar dilution,MTS,Vitek (GN16) with respect to broth microdilution.The CA is 79.6％ (82/ 103,S≥14 mm,R≤10 mm),69.9％ (72/103,S≥ 16 mm,R≤ 12 mm),34.0％ (35/103,S≥19 mm,R≤14 mm)using disk diffusion method compared with broth microdilution (FDA break point criteria).Conclusions The susceptibility of tigecycline must be tested using fresh medium.The medium brands and lot numbers used in this test have no effects on the tigecycline susceptibility to Acinetobacter baumannii and Klebsiella pneumoniae.There exist the better correlations on MIC using agar dilution and MTS than the disk diffusion and Vitek(GN16) compared with broth microdilution.It is expected that the consistency can be improved by adjusting the break point of disk diffusion.

It has been known that the Alzheimer's disease(AD)is related closely with a synaptic failure,and the p21-activated kinase(PAK)is well documented to play an important role in the regulation of the synaptie functions.However,the relationship between thePAK and the pathology of AD is unclear.In the present study,we examined the expressions of the PAK3(one subtype ofPAK),phospho-rylated-PAK(pPAK) and β-amyloid42(Aβ42,β-amyloid with 42 peptides)in an APP/PS1 double transgenie mouse model of AD andthe morphologies of geurOtlS in the hippocampus at different ages.The Western Blot results showed that the expression of PAK remainedunchanged,while,the expression of pPAK decreased largely at the age of 32 weeks and further decreased significantly with aging in thehippocampus of the APP/PS1 transgenic mouse.A1342 levels in the hippocampus were detected to increase as early as the age of 22 weeks,and kept the increase to continue with aging.The morphological results showed no obvious neuron loss in the sections of Nissl staining,while serious distonion and disorder of the dendrites of the hippocampal neurons were observed on the sections of Gelgi staining in theAPP/PS1 transgenic mouse.The present results suggested that it seemed something wrong in the processes of phospholization of PAK,butnot in the expression of the PAK itself;the toxic Aβ42 might affect the PAK in its phospholization,which in turn directly influence thedendritic development in the hippocampal neurons and cause the dendrites distorting and disordering.

Objective:To investigate the ethics and existing problems of experimental animals in medical experiments and experimental teaching, and to formulate countermeasures so that animal ethics and animal welfare can be truly reflected in medical experiments.Methods:In this study, a "Basic Function Experiment Center Animal Experiment Questionnaire" with 25 questions was formulated from three aspects: the ethical cognition of experimental animals, whether animal experiments are ethical or not, how to view the problems of animal ethics and experimental teaching and the cognition of virtual simulation experiment teaching. Questionnaire was sent to Hubei University of Medicine to investigate the international students of Batch 2017 (5-year program), undergraduates of Batch 2017 (5-year program) and nursing students of Batch 2018 (4-year program) as well as teachers, researchers and employees of laboratory animal center (all with bachelor degree or above). The survey results were expressed as percentage.Results:The recovery rate of the questionnaire in this study was 98.04%(2 451/2 500), among which the practitioners, teachers and researchers in the laboratory animal center clearly understood the ethics of experimental animals, but there was a widespread phenomenon of lagging ethics among the students. For example, 16.24% (398/2 451) students had not received animal experiment ethics education and training, 29.46% (722/2 451) were not clear about animal protection laws and regulations, 7.14% (175/2 451) thought animal experiments were immoral; 29.54% (724/2 451) had vague cognition of animal welfare and ethical knowledge; 25.91% (635/2 451) were not familiar with the operation steps; 9.38% (230/2 451) were indifferent to the extra treatment of animals due to operation errors, 7.83% (192/2 451) chose to give up the experiment in the treatment of animals after massive bleeding, only 5.43% (133/2 451) chose to continue the experiment after timely hemostasis and infusion, and 9.26% (227/2 451) chose to do operations unrelated to the experiment. After the experiment, 2.28% (56/2 451) chose to kill the animals by bloodletting, only 5.51% (135/2 451) chose excessive anesthesia euthanasia, and 1.96% (48/2 451) chose to kill the animals by cervical dislocation and violence. Only 15.79% (387/2 451) chose to remember the dead animal for 2 minutes. Only 32.56%(798/2 451) of the respondents understood virtual simulation experiment, 34.92% (856/2 451) of the respondents thought that virtual simulation experiment or experimental teaching video could be used to replace the existing live animal experiment, 77.56% (1 901/2 451) believed that the construction of virtual simulation laboratory should be strengthened.Conclusion:It is imperative to strengthen the education of students' ethics of experimental animals, which is conducive to the establishment of correct ethics of experimental animals for medical students, so that the "3R" principle and animal welfare can be truly implemented in experimental teaching and scientific research experiments.