Aim To evaluate the effect of expression inhibition of spermine oxidase(SMO)on the actitumor activity of polyamine analogue CPENSpm (N~1-cyclopropylmethyl-N~(11)-ethylnorspermine).Methods siRNA technique was used to inhibit expression of SMO in human lung cancer line A549.QT-RT-PCR and enzyme activity assay was performed to determine the expression level of SMO.The cell proliferation was detected by MTT assay.The apoptosis of A549 cells were evaluated by DNA degradation and Sub-G_1/flow cytometry assay.Results The A549 cell line with silenced SMO expression was successfully obtained.Basic SMO mRNA and enzyme activity levels in the SMO-siRNA plasmid transfected cells were 0.53% and 14% lower than that in the control cells respectively. Treating A549 control cells by 10 μmol·L~(-1) CPENSpm for 24 hours resulted in a 10-folds up-regulation of SMO in mRNA level and 20-fold increase in enzyme activity,but this drug-induced SMO expression was obviously prevented in SMO-siRNA plasmid transfected cells.MTT assay demonstrated that SMO expression inhibition decreased the sensitivity of A549 cells to CPENSpm exposure(0～20 μmol·L~(-1)).DNA degradation and sub-G_1 assay proved a deceased ability of CPENSpm to induce apoptosis in SMO-siRNA plasmid transfected cells.Conclusion Up-regulation of SMO by CPENSpm is possibly one of the molecular basics for its antitumor activity.

Objective To investigate whether difluromethylornithine (DFMO) can be used in the treatment of human leukemia. Methods The cell proliferation was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)] assay after treatment of human lymphocyte Jurkat cells by DFMO (0 to 10 mmol/L) for 24 to 72 h. Enzyme activity of spermine oxidase (SMO) and acetylpolyamine oxidase (PAO) was determined by chemiluminesence assay. DNA fragmentation assay was used to evaluate cell apoptosis. Fluorescent dye assay was performed to determine the changes in mitochondrial membrane potential. Western blotting was used to determine Bax content. Casepase-3 enzyme activity was measured by spectrophotometric method. Results DFMO treatment inhibited the proliferation of Jurkat cells significantly in a dosage- and time-dependent manner (P

AIM: To prepare recombinant human spermine oxidase (SMO) and polyclonal antibody against human SMO by gene recombination techniques. METHODS: Human SMO cDNA was amplified from total RNA of A549 cells through reverse transcription PCR. The cDNA was then cloned into pET-15b to construct SMO prokaryotic expression vector. After transforming, the vector was induced to express recombinant SMO by IPTG in E. coli BL21 (DE_3). Recombinant SMO was purified by Ni-NTA resin under denaturing condition and then was dialyzed to renature. The enzyme activity of recombinant SMO was analyzed by chemical fluorescent method. SMO polyclonal antibody was prepared by using recombinant human SMO protein purified by polyacrylamide gel electrophoresis as antigen to inoculate rabbit intradermally. The titer and specificity of anti-sera were determined by ELISA, Western blot and Immune Cell Chemistry. RESULTS: Purified and dialyzed recombinant human SMO has the specificicity of oxidizing the spermine. The polyclonal antibody has high titer and specificity against human SMO. CONCLUSION: This research established a method for prokaryotic expression, purification and polyclonal antibody preparation of human SMO. The method lays a foundation for the future functional research of SMO.

Objective To observe any therapeutic effect of laser irradiation on pathological inflammatory reactions in ulcerative colitis (UC) induced by oxazolone,and to investigate possible mechanisms.Methods Six rats were selected as a normal control group.Another 24 rats with UC induced by oxazolone were randomly assigned to a UC model group (n =8),a 400 mW laser group (161.3 mW/cm2,n =8),and a 200 mW laser group (80.6 mW/cm2,n =8).All the rats were fixed in custom-built devices.Those in the therapy groups were treated daily,10 min per time,for 10 days.After the end of the last irradiation session,disease activity indexes (DAIs)were observed.Rats from every group were sacrificed 24 h after the last irradiation in order to observe any pathological changes in colon tissue,the weight of fresh ulcerated tissues,and gross changes in morphology.Colon segments were stained with hematoxylin-eosin and histological lesion scoring was performed under a light microscope.Any changes in inflammatory edema in the colonic mucous membrane were observed before and after laser irradiation.Results The UC model was successfully established.Average body weight in the 400 mW laser group increased significantly more than in the UC model group,approaching that of the normal control group.DAI decreased significantly.The thickness of the epithelial mucous membrane,lamina propria and submucosa was basically restored.Histological lesion scores also improved significantly,and the weight of fresh ulcerative tissue was significantly lower.Mucous membrane ulcers,submucosa edema and inflammatory cell infiltration all were alleviated significantly,merely presenting a few inflamed cells and small amounts of periphlebitis.In the 200 mW laser group all these outcome measures improved significantly compared with the UC model group,but were not as good as in the 400 mW laser group,and the rats needed longer to recover.Conclusion Laser irradiation at 400 mW has advantages over 200 mW,and could significantly relieve the pathological inflammatory response in colonic tissue,decrease submucosa edema and ameliorate other symptoms of ulcerative colitis,at least in rats.

Objective To explore the effects and molecular mechanisms of laser therapy on serum and colon tumor necrosis factor α (TNF-α),interleukin-6 (IL-6) and interleukin-10 (IL-10) in rats with oxazolone induced ulcerative colitis (UC).Methods Thirty adult male SD rats were randomly divided into four groups:a normal group ( n =6),a UC model group ( n =8 ),a 400 mW laser treatment group ( n =8 ) and a 200 mW laser treatment group ( n =8 ).Odified oxasolone sensitization was used to induce UC models in the rats.The AsAlGa semiconductor laser used in the treatment had a power of 200 mW or 400 mW.The therapy lasted for 10 days with daily 10 min sessions.The rats were sacrificed after treatment and enzyme-linked immunosorbent assay (ELISA) was used to measure IL- 6,IL- 10 and TNF-α in serum and ulcer tissues in all groups.Results Compared with the normal group,the weights of the UC model rats were significantly lower,and they had severe mucoid bloody stool.TNF-α and IL-6 levels in serum and colon tissues increased significantly and IL-10 decreased significantly,so the UC model was successfully established.After laser treatment,body weight gain and stool moderation in rats were observed.in the 400 mW group,and average TNF-α and IL-6 levels in both serum and colon tissue decreased significantly.IL-10 increased significantly,close to the level of the normal group.In the 200 mW group,serum TNF-α and IL-6 levels were significantly lower,and IL-6 in the colon tissue was reduced significantly.TNF-α did not lessen significantly,and serum and colon tissue IL-10 levels did not improve significantly compared with the model group.Conclusions 400 mW AsAlGa semiconductor laser irradiation can effectively regulate cytokines after oxaZolone induced UC in rats in two ways.It reduces pro-inflammatory cytokines and increases the level of anti-inflammatory cytokines.This may imply one of the underlying molecular mechanisms of low energy laser treatment of UC.

Objective The study is designed to evaluate the difference in identification for depression and clinical effectiveness of its treatment between psychiatrists and non-psychiatric Dhysieians at out-patient departments of general hospitals and to analyze its related factors.Methods Totally,680 patients who visited psychiatric clinics in nine general hospitals of Shanghai at first time were screened by psychiatrists using Composite International Diagnostic Interview(CIDI)for depression and the screening resuhs were compared to the diagnosis made by non-psychiatric physicians as goal-keepers there.Altogether 297 patients with depression were recruited and assessed using Hamilton Depression Rating Scale(HAMD)and Hamilton Anxiety Rating Scale(HAMA).A self-made questionnaire for basic information was used to assess the ability for identification of depression in non-psychiatric physicians of general hospitals.Results Psychiatrists identified depression correctly in 337 of 680 patients who visited psychiatric clinics in general hospitals at their first visit,but non-psychiatric physicians only identified 216 patients,with statistically significant difference(χ2=30.73,P=0.000).A consistent agreement on depression diagnosis between the findings by psychiatrists and by CIDI was reached with Kappa of 0.774,but Kappa for that between the findings by non-psychiatric physicians and CIDI was 0.439.Effectiveness of treatment for depression by psychiatrists was better than that by non-psychiatric physicians,with higher total score and scores for each item of HAMD four,eight and 12 weeks after treatment,respectively(P＜0.05 or P＜0.01).Length of professional work of non-psychiatric physicians and annual time for professional training in management of mental disorders associated with their ability of identification for depression(P＜0.05).Conclusion Ability of non-psychiatric physicians to identify alQd cope with depression in the psychiatric department of general hospitals was insufficient,suggesting that more importance should be attached to the management of service quality at psychiatric department of general hospitals.

Objective:To analyze the protecting role of facial nerve motor evoked potential (FNMEP) monitoring in facial nerves during the resection of cerebellopontine angle lesions.Methods:The clinical data and monitoring data of 83 patients with cerebellopontine angle lesions, accepted intraoperative facial nerve motor evoked potential (FNMEP) monitoring in our hospital from January 2019 to December 2019, were retrospectively investigated. Pre-surgical (opening dural), intraoperative and post-surgical (closing dural) FNMEP amplitudes were recorded. Relation of end (closing dura) to start (opening dura) amplitude ratio with facial nerve function grading at early-term (3 d after surgery) by House-Brackmann (H-B) grading was analyzed.Results:In these 83 patients, 75 patients (90.4%) achieved total tumor resection, and 8 patients (9.6%) achieved subtotal resection. Integrated anatomical preservation of facial nerves was achieved in all patients (100%); one (1.2%) was died for postoperatively delayed hemorrhage. Three d after surgery, facial nerve H-B grading I-II was noted in 62 patients, and the end to start amplitude ratio was 85.3% (66.8%, 93.4%); facial nerve H-B grading III was noted in 11 patients, and the end to start amplitude ratio was 45.6% (38.7%, 43.8%); facial nerve H-B grading IV was noted in 7 patients, and the end to start amplitude ratio was 23.1% (16.4%, 25.6%); facial nerve H-B grading V-VI was noted in 3 patients, and the end to start amplitude ratio was 6.7% (3.5%, 7.7%). There was a negative correlation between end to start FNMEP amplitude ratio and post-surgical early HB grading ( r s=-0.895, P=0.000). Conclusion:FNMEP is highly reliable in predicting early postoperative facial function during resection of cerebellopontine angle lesions, which can be a valid protection technique for facial nerve.

During the Yan’an period, under the instructions of the CPC Central Committee and the Central Military Commission, the former Eighth Route Army Health School was expanded into China Medical University. As the first medical university founded by the Communist Party of China, this is a great practice of higher medical education in the new revolution. The school adhered to the policy of educating people with both ability and political integrity, the "saving the dying and healing the wounded, implementing revolutionary humanitarianism" has become the ideological guide and goal of the majority of revolutionary medical workers. The school provided a good teaching environment by strengthening teachers, enriching infrastructure and teaching equipment. Teachers and students have overcome difficulties and made great achievements in teaching and learning. At the same time, through the mass production movement and rectification movement, the development of the school was promoted from two aspects of material production and ideological construction. The practice of China Medical University in education policy, teaching curriculum and teaching content has certain enlightenment for today’s medical education.

Objective: To discuss the affect of glycosylated hemoglobin (HbA1c) level for the onset of nonalcoholic fatty liver disease (NAFLD) in cohort population. Methods: An epidemiological survey of the relationship between HbA1c and NAFLD conducted in 2012 was based at cohort baseline, and three follow-up sessions conducted in 2013, 2014 and 2015. In total 2 811 subjects were included in the study after exclusion of NAFLD patients at baseline and those who lost their lives due to relocation, and death. The Cox proportional hazard model was used to analyze the relationship between glycosylated hemoglobin and other risk factors of NAFLD. Continuous variables were compared using the t-test or the Mann-Whitney test. χ ²-test was used for the measurement of categorical data. Results: A total of 2 811 subjects with mean age of 59 (58.2±9.8) years old, including 1 664 males and 1 147 females. Age, waist circumference, body mass index, systolic blood pressure, γ-glutamyltransferase and fasting blood glucose level of HbA1c abnormal group were higher than normal group. The incidence of NAFLD in the abnormal HbA1c level group (25.4%) was higher than normal group (14.9 %), and diastolic blood pressure, high-density lipoprotein cholesterol was lower than normal group and the differences were statistically significant. During the three follow-up intervals, there were 440 new cases of NAFLD, consisting 285 males and 155 females with cumulative incidence of 15.7% (440/2 811). Multivariate Cox regression analysis showed that patients with elevated HbA1c had a higher risk of developing NAFLD (HR 1.796; 95% CI 1.335~2.418; P < 0.01), and the increased HbA1c level after adjustment for gender, age, and metabolic syndrome-related factors remained an independent risk factors for NAFLD (HR 1.580; 95.0% CI 1.161-2.152; P < 0.01). Conclusion An elevated HbA1c levels have a positive predictive value for the onset of NAFLD.

Mutations or inactivation of parkin, an E3 ubiquitin ligase, are associated with familial form or sporadic Parkinson's disease (PD), respectively, which manifested with the selective vulnerability of neuronal cells in substantia nigra (SN) and striatum (STR) regions. However, the underlying molecular mechanism linking parkin with the etiology of PD remains elusive. Here we report that p62, a critical regulator for protein quality control, inclusion body formation, selective autophagy and diverse signaling pathways, is a new substrate of parkin. P62 levels were increased in the SN and STR regions, but not in other brain regions in parkin knockout mice. Parkin directly interacts with and ubiquitinates p62 at the K13 to promote proteasomal degradation of p62 even in the absence of ATG5. Pathogenic mutations, knockdown of parkin or mutation of p62 at K13 prevented the degradation of p62. We further showed that parkin deficiency mice have pronounced loss of tyrosine hydroxylase positive neurons and have worse performance in motor test when treated with 6-hydroxydopamine hydrochloride in aged mice. These results suggest that, in addition to their critical role in regulating autophagy, p62 are subjected to parkin mediated proteasomal degradation and implicate that the dysregulation of parkin/p62 axis may involve in the selective vulnerability of neuronal cells during the onset of PD pathogenesis.
Adaptor Proteins, Signal Transducing ; chemistry ; metabolism ; Animals ; HEK293 Cells ; Heat-Shock Proteins ; chemistry ; metabolism ; Humans ; Lysine ; metabolism ; Mice ; Neurons ; metabolism ; pathology ; Oxidopamine ; pharmacology ; Parkinson Disease ; metabolism ; pathology ; Proteasome Endopeptidase Complex ; metabolism ; Protein Stability ; Proteolysis ; drug effects ; Sequestosome-1 Protein ; Ubiquitin-Protein Ligases ; metabolism ; Ubiquitination ; drug effects