Data were analyzed from self-filled questionnaires for the students who took the elective course of general practice in China Medical University. 95.5% of students thought it was necessary to study general practice in the college. 0-10 was used to score the degree of how they want to choose general practice as a career, the median was 0 before they had the class and the median was 5 after the class (Wilcoxon rank test, P<0.01). But there were still some places need to be improved in terms of curriculum design, teaching methods and community practice. The General Practice teaching need to be innovated and the teaching mode should be different from the traditional one.

In this paper, the effect of 5% (V/V) n-alkanes (e.g, n-Heptane, n-Octane, n-Decane, n-Dodecanen-Tetradecane and n-Hexadecane) on the growth and protease production of organic-solvent-tolerant- bacte-rium Bacillus licheniformis YP1 was studied. 5%(V/V) n-alkanes had no effect on the stability of YP1 prote-ase. 5% (V/V) n-alkanes had no notable influence on the yield of strain YP1 but dramatically affected theprotease production. The presence of n-Heptane, n-Octane and n-Decane deeply repressed the protease pro-duction; however n-Dodecane, n-Tetradecane and n-Hexadecane enhanced the protease production promi-nencely. The concentration of n-Tetradecane (1%-8%, V/V) had a direct ration with the protease production.The detailed experiments showed that the notable increase of protease activity appeared at the late logarithmof cultivation compared with the blank. The cell shape of YP1 strain remarkably decreased when grown inthe presence of n-Tetradecane. This is the first report about the effect of n-alkanes on the protease productionby the solvent tolerant bacterium.

For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVETo investigate the therapeutic effect of IL-6 mAb on experimental autoimmune myocarditis (EAM) in rats, and search the mechanism of the role of IL-6, helper T cells 17 (Th17) and regulative T cells (Treg) in EAM pathogenesis.

METHODSThirty-four Lewis rats were divided into three groups randomly, i.e. control group (n = 6), EAM group (n = 12), and IL-6 mAb intervention group (n = 16). Rats in EAM group and IL-6 mAb intervention group were injected intracutaneously with myosin to establish EAM model. Rats in IL-6 mAb intervention group were injected intraperitoneally with 1 mg IL-6 mAb on 1st, 7th to 20th day after cardiac myosin immune injection. Myocardial inflammation was examined by HE stain, Masson stain, and TdT assay (TUNEL reaction) on 21st and 84th day after IL-6 mAb therapy in order to assess the therapeutic role. Spleen cells were analyzed by flow cytometry to illustrate Th17 and Treg cells? number and function. The serum concentration of IL-6, IL-10, IL-17, and TGF-beta in each group was measured by ELISA, concentration of STAT3, RORgammat, and Foxp3 mRNA in each group was determined with RT-PCR. Spleen cells derived from EAM were stimulated by IL-6 mAb in vitro, and the concentration of IL-10, IL-17 and TGF-beta was measured by ELISA.

RESULTSInflammation score, fibrosis score, and apoptosis index in IL-6 mAb intervention group were significantly decreased as compared with those in EAM group (P < 0.01). The number of Th17 and Treg cells in EAM group on the 21st day (experimental acute peak stage) were increased, and those in intervention group on the 21st day were significantly inhibited (P < 0.01). The concentration of serum IL-6, IL-10, IL-17 and TGF-beta in intervention group on the 21st day was decreased dramatically in comparison with that in EAM group on the same day (P < 0.01). The levels of peripheral blood STAT3, RORgammat, Foxp3 mRNA in intervention group on the 21st day was decreased significantly as compared with that in EAM group (P < 0.01). The expression of IL-10, IL-17 and TGF-beta was increased significantly (P < 0.01) by stimulation of IL-6 mAb on spleen cells derived from EAM in vitro.

CONCLUSIONSIL-6 mAb could neutralize IL-6, and ameliorate myocarditis and reduce heart autoimmune responses. IL-6 mAb has significantly protective effects on EAM by suppressing Th17 and Treg cells.

Animals ; Antibodies, Monoclonal ; therapeutic use ; Autoimmune Diseases ; drug therapy ; immunology ; Disease Models, Animal ; Forkhead Transcription Factors ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-17 ; metabolism ; Interleukin-6 ; immunology ; Male ; Myocarditis ; drug therapy ; immunology ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; metabolism ; Rats ; Rats, Inbred Lew ; STAT3 Transcription Factor ; metabolism ; Th17 Cells ; immunology ; Transforming Growth Factor beta1 ; metabolism

Objective To investigate the expression of c-FLIPL in leukemia and explore its clinical significance. Methods The expression level of c-FLIPL mRNA in bone marrow mononuclear cells was determined by real-time polymerase chain reaction(PCR)in 103 leukemia patients with different types of leukemia,including 54 cases of acute lymphocytic leukemia(ALL)with 37 newly diagnosed,5 relapsed,and 12 complete remis-sion,38 cases of acute myeloid leukemia(AML)with 24 newly diagnosed,6 relapsed,and 8 complete remission,newly diagnosed 2 cases of acute undifferentiated leukemia(AUL),6 cases of chronic myelocytic leukemia(CML),and 3 cases of chronic myelomonocytic leukemia(CM-ML). The immunophenotype of patients were detected by flow cytometry. Results Expression level of c-FLIPL mRNA was higher in newly diag-nosed and relapsed leukemia patients. There was no significant difference between newly diagnosed and relapsed leukemia patients(P>0.05). Ex-pression level of c-FLIPL mRNA of AUL and CML was higher than that in other patients ,but there was no significant difference(P>0.05). Ex-pression level of c-FLIPL mRNA of all newly diagnosed and relapsed leukemia patients was significantly higher than that in control group and com-plete remission group(P<0.05). The expression level of c-FLIPL mRNA was correlated with risk stratification ,white blood cell(WBC),lactate dehydrogenase(LDH),hydroxybutyrate dehydrogenase(HBDH),CD45 and TEL-AML1,but was not associated with age,sex,fibrinogen and chromosome abnormality. Conclusion c-FLIPL mRNA is highly expressed in leukemia patients ,and is closely related with risk stratification , WBC,LDH,HBDH and prognosis.

Objective: To observe the effects of moxibustion therapy at Shenshu (BL 23) and Zusanli (ST 36) of rats with induced rheumatoid arthritis (RA) on the serumal intercellular cell adhesion molecule-1 (ICMA-1) and the pathological tissue, to discuss the mechanism of the warming and activating effect of moxibustion. Methods: After establishing the RA rats model, the induced rats were treated with moxibustion therapy on the acupoint Shenshu (BL 23) and Zusanli (ST 36), followed by analyzing the pathological section of the ankle of the hind limb and testing the ICAM-1 content with ELISA. Results: The plantar circumferences of the induced rat increased significantly compared with the rats in the control group (P<0.01), accompanying with the increase of the synovial layer, the erosion of phlogocytes to chondrocytes and the specific increase of ICAM-1 content. After the moxibustion therapy, the plantar circumferences decreased significantly (P<0.01) while the synovial layer tended to reduce. In addition, there was no pathological damage of the articular cartilage and the ICAM content decreased with significant deviation (P<0.01), compared to the model group. Conclusion: It was concluded that moxibustion therapy could inhibit the arthrosynovitis and hyperplasia, ameliorate the erosion of phlogocyte to cartilage, prevent articular periosteal lesions and delay the pathological course. The warming and activating effect of moxibustion therapy may involve the inhibition of the formation of ICAM-1 and pannus.

Objective To establish a high throughput screening assay for identifying human small molecular antagonists targeted IL-6R.Methods The full length gene of the human IL-6R extracellular region was amplified by PCR and cloned into a eukaryotic expression vector to construct recombination expression plasmid pABHis -IL6R that was then transfected transiently into HEK293T cells to prepare recombination protein IL-6R.Western blotting assay and receptor-ligand binding experiment were used to analyze the bioactivity of IL-6R.A new screening method based on ELISA was established using the function of IL-6R binding to its ligand and the characteristics of Fc fragment binding to IgG-HRP.Then Z′-factor was calculated and a known antagonist ab 47215 was used to assess the stability and reliability of the new assay .Results Recombination plasmid pABHis-IL6R was constructed and soluble IL-6R was prepared.IL-6R reported herein could be recognized by an anti-IL-6R antibody and specifically bind to its ligand in a dose response manner .A Z′-factor of 0.53 was obtained that could serve high throughput screening assay .Ab47215 , as a known specific antagonist , was able to block rhIL-6 from binding to the receptor in a dose-dependent manner in the new screening assay , the IC50 of which was (0.55 ± 0.11)μg/ml.Conclusion An innovative and easy screening assay for identifying human IL-6R antagonists is established , which might help discover potent and specific antagonists .

One hundred and forty-four medical students were enrolled in the study, the participants were randomly divided into experimental group and control group with 72 in each. All students attended the theoretical lecture of general medicine. Students in experimental group took part in the extracurricular teaching activities including the nutritional course and practice, the clinical case study and problem based learning ( PBL), students in control group did not have the extracurricular activities. The results showed that although there was no difference in general testing scores between two groups, the problem solving ability and assay-writing ability of experimental group were significantly higher than those of control group.

Objective:To observe the effects of moxibustion on serum levels of interleukin (IL)-6, IL-8 and tumor necrosis factor-α (TNF-α), and to explore the effects of moxibustion on inflammatory damaging factors in experimental rheumatoid arthritis (RA) model rats; the relationship between the therapeutic effect of moxibustion on RA and the change in the Toll-like receptor (TLR) signaling pathway was analyzed using Toll-like receptor 4 (TLR4) antagonists and agonists. Methods:Fifty Sprague-Dawley (SD) rats were divided into a normal group, a model group, a moxibustion group, a moxibustion plus TLR4 agonist group (agonist group) and a moxibustion plus TLR4 antagonist group (antagonist group) according to the random number table, with 10 rats in each group. Except the normal group, rats in the other four groups were subjected to model preparation with the wind, cold and wet environmental factors plus Freund's complete adjuvant (FCA). Rats in the normal and model groups were not treated; rats in the moxibustion, agonist and antagonist groups started to be treated with the moxibustion (cigarette-type moxa) at bilateral Shenshu (BL 23) and Zusanli (ST 36) from the 4th day after the successful modeling, for 20 min each time with a total of 10 d. Rats in the agonist and the antagonist groups were injected with TLR4 agonist or antagonist [0.1 mg/(kg·bw)] via the tail vein 30 min before moxibustion. The concentrations of serum IL-6, IL-8 and TNF-α in each group were determined by enzyme-linked immunosorbent assay (ELISA). Results:Compared with the normal group, in the model group, the rat's right hind paw swelling was significantly obvious (P<0.01), there was a lot of inflammatory infiltration in the synovial tissues, the surface of the synovial membrane was unsmooth, the synovial membrane was hyperplasia and thicker, and the serum IL-6, IL-8 and TNF-α concentrations increased significantly (P<0.05). Compared with the model group, the paw swelling degrees of the rats in the moxibustion, the agonist and the antagonist groups reduced significantly (allP<0.01); the swelling degree in the antagonist group was milder than that in the agonist group, but the between-group difference was not statistically significant (P>0.05); inflammatory infiltration and synovial membrane hyperplasia in the synovial tissues of the moxibustion group and the antagonist group were all relieved differently; the decrease of synovial layer number in the moxibustion group was more obvious, and there were no obvious improvements in inflammatory infiltration and synovial thickness in the agonist group; the concentrations of IL-6, IL-8 and TNF-α in the moxibustion group were decreased, and the differences in the IL-6 and TNF-α concentrations were statistically significant (allP<0.01); there was no significant between-group difference in the IL-8 concentration (P>0.05); the concentrations of serum IL-8 and TNF-α in the agonist group increased significantly (both P<0.01), while the IL-6 concentration decreased without significant difference (P>0.05); the concentrations of IL-6 and IL-8 in the antagonist group decreased but the between-group differences were statistically insignificant (bothP>0.05), and the TNF-α concentration significantly increased (P<0.05). Compared with the moxibustion group, IL-6, IL-8 and TNF-α concentrations increased in the agonist group, and the differences in the IL-8 and TNF-α concentrations were statistically significant (both P<0.01); the concentrations of IL-6, IL-8 and TNF-α increased in the antagonist group, and the differences in the IL-6 and TNF-α concentrations were statistically significant (bothP<0.01); there was no significant difference in the IL-8 concentration between the groups (P>0.05). The serum levels of IL-6, IL-8 and TNF-α in the antagonist group were lower than those in the agonist group (allP<0.05). Conclusion:Moxibustion at Shenshu (BL 23) and Zusanli (ST 36) can reduce the joint swelling degree and inflammation in synovial tissue of RA model rats, decrease the serum levels of IL-6, IL-8 and TNF-α in RA model rats; the decreases of IL-6 and TNF-α are more significant than the decrease of IL-8; TLR4 agonist and antagonist can significantly attenuate the effect of moxibustion in inhibiting releases of IL-6, IL-8 and TNF-α, so that the change in TLR signaling pathway affects the effect of moxibustion in inhibiting the releases of IL-6, IL-8 and TNF-α.

Objective: To understand the incidence of school bullying and its influencing factors among adolescents in low-income and middle-income countries, and to explore the association between school bullying and mental health of adolescent students, so as to provide reference for prevention and control of school bullying and mental health intervention. Methods: Data was obtained from the 2009-2015 Global School Student Health Survey from 19 low-income and middle-income countries (n=22 963). Binary Logistic regression was used to analyze the influencing factors of school bullying, and multiple linear regression was used to analyze the relationship between school bullying and mental health. Results: The average score of students mental health was(5.75±2.09), and approximately 35.1% of adolescent students reported suffering from school bullying. The rates of school bullying among students in low-income and middle-income countries were 39.4% amd 34.3%, respectively. Students with lower grades, overweight, poor family economic status, low family learning and psychological support, poor perceived family relationship, more truancy, and poor relationship with classmates were more likely to suffer from school bullying(P<0.05). Exposure to school bullying was positively associated with adverse mental health outcomes for women(B=1.27, P<0.01). Conclusion Not only were school bullying more common in low-income countries, but also school bullying had a greater negative impact on the mental health of girls. We need to pay more attention to school bullying among adolescent students, especially in low-income countries and girls, with cost-effective interventions to reduce or mitigate the consequences of bullying.