In the reform of medical microbiology education on medical students of 8-year-education pro-gram, we compared the traditional teaching methods with Problem Based Learning (PBL) method. Through our practice, we have found that the combination of traditional lecture-based learning and PBL seems to better match the students’ way of learning. The lack of basic knowledge of the students hindered their learn-ing effect during the bilateral discussion in the PBL education. We also found that the application of PBL in medical microbiology education is an iterative process and should be promoted step by step. The theoretical level and the innovative ideas of the teachers play a crucial role in the dynamic process of education reform.

ObjectiveTo investigate effects ofEuphorbiae ebracteolatae Radixon the proliferation and apoptosis in human papillomavirus (HPV)-18-infected HeLa cells.MethodsThe alcohol extract ofEuphorbiae ebracteolatae Radixwas prepared. Cell proliferation was determined using the MTT assay, and apoptotic cells was determined using the Hoechst 33258 staining.ResultsHPV-18-infected HeLa cells were treated with 10, 20 μg/mlof the alcohol extract ofEuphorbiae ebracteolatae Radixfor 12 h, the cell proliferation inhibition rate were significantly increased compared to the control group (16.36% ± 5.06%, 18.59% ± 5.99%; allP<0.01);HPV-18-infected HeLa cells were treated with 0.625, 1.25, 2.5, 5, 10, 20 μg/ml of the alcohol extract ofEuphorbiae ebracteolatae Radixfor 72 h, the cell proliferation inhibition rate were significantly increased compared to the control group (13.82% ± 3.66%, 30.24% ± 6.58%, 43.29% ± 4.29%, 53.32% ± 1.41%, 54.24% ± 1.97%, 50.68% ±1.74%;allP<0.01). The Hoechst 33258 staining showed that HPV-18-infected HeLa cells were treated with 10 μg/ml of the alcohol extract ofEuphorbiae ebracteolatae Radixfor 24 h, the apoptotic cells were found.ConclusionEuphorbiae ebracteolatae Radixcould inhibit the proliferation and induce apoptosis in HPV-18-infected HeLa cells.

Objective To explore the mechanism of blood-stage treatment in patients with psoriasis vulgaris through studying the level change of IFN-?, IL-4, IL-10 and IL-17 in patients with blood-stage treatment during activity and quiescence period separately. Methods 20 patients with psoriasis vulgaris in active stage(blood heat syndrome)and 20 patients with psoriasis vulgaris in resting stage(blood stasis syndrome) were recruitedto observe the treatment effects by the PASI score,and to observe thechange of IFN-?, IL-4, IL-10, IL-17 before and after treatment in the serum by ELISA. Results The PASI scores of two groups were both significantly decreased after treatment (blood heat syndrome group t=6.685, P<0.01;blood stasis syndrome group t=4.959, P<0.01). The levels of cytokines were significantly different between patients in the periods of activity and quiescence. Onvarying degrees, the levels of cytokines of two groups were improved after treatment. The levels of cytokines IFN-?, IL-17 in blood heat group significantly decreased(t=3.024, P<0.01;t=2.543, P<0.05). The levels of cytokines IL-17 drop but the levels of cytokines IL-4 raised in blood stasis group,that were significantly differentwith the levels before treatment(t=2.417, P<0.05; t=2.547, P<0.05). Conclusion The levels of INF-?, IL-4, IL-10 and IL-17 could be effectively modulated with blood-stage treatment in treating psoriasis vulgaris.

To obtain high-yield avilamycin-producing strains,low energy N~+ ion implantation technology and screening of streptomycin-re- sistant mutants are used in the study on breeding mutation.The results show that,“saddle”region,which range is from 3?10~(15) to 5?10~(15) ions/cm~2,has got better induced mutation action.It also means that the strain's resistant mutation and yield mutation closely correlate to each other,and the method of streptomycin resistant screening is feasible.We have isolated a high-yield strain SVT-45 which the productivi- ty is 195% higher than the original strain's in the rotation-flask experiments.These results showed that the ion implantation was an effective method for microbe mutagensis.

Background Posterior capsular opacification(PCO) is common complication after extrecapsular extract of cataract.Matrix metalloproteinases-3 (MMP-3) can degrade all the extracellular matrix except polyose.The gene therapy of PCO upon MMP-3 is the researching hot topic.Fibronectin ( FN ) is a degrade gelatin,so its expression can reflect the effect of MMP-3 on LECs indirectly. Objective The aim of this study was to construct MMP-3 eukaryotic recombination plasmid and transfect to lens epithelium cells(LECs) for the observation of MMP3 expression,and to explore the feasibility of gene therapy for after cataract. Methods Six fresh lenses were obtained from pigs.LECs were cultured using explant method.The eukaryotic expression vector pEGFP-N1-MMP-3 was reconstructed with MMP-3 and pEGFP-N1 plasmids.The accuracy of MMP-3 gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting pEGFP-N1-MMP-3 into LECs of pig,the expression of MMP-3 protein in the cells was indirectly observed by green fluorescent protein.The expression of FN in LECs was detected using Western blot. Results The result of double enzyme digestion was consistent with the base number of pEGFP-N1 plasmids and target fragment.By enlacing the result of DNA sequencing analysis with software,the resemblance of the DNA sequence of MMP-3 from recombination plasmid pEGFP-N1-MMP-3 and that of homo MMP-3 was 99.6％,indicating that the target fragment was inserted to pEGFP-N1 plasmids successfully.Green fluorescence for GFP was seen in the LECs in pEGFP-N1-MMP-3 transfected group,but absent response for GFP was in empty vector group.Western blot revealed that the relative expression level of FN in LECs was 0.666±0.008 in pEGFP-N1-MMP-3 trasfected group and 0.326 ±0.071 in empty vector group,with a significant difference between these two groups(P=0.000). Conclusions Eukaryotic recombination plasmid pEGFP-N1-MMP-3 is successfully constructed,and MMP-3 can be expressed in LECs after transfected.These results lay a foundation for the further research of MMP-3 gene therapy for PCO.

OBJECTIVETo evaluate efficacies of three commonly used oral drugs including Berbamine Hydrochloride Tablet (B), Qijiao Shengbai Capsule (Q), and Leucogen Tablet (L) (by single drug, two drugs or three drugs) combined with granulocyte colony-stimulating factor (G-CSF) for treat ment of chemotherapy related leukocytopenia in mice.

METHODSTotally 156 Kunming male mice were divided into the normal control group (A, n=24), the model group (B, n=24), the G-CSF group (C, n =24), the G-CSF+Q group (D, n=12), G-CSF+ B (E, n=12), the G-CSF+L group (F, n=12), the G-CSF + Q + B group (G, n=12), the G-CSF + Q + L group (H, n=12), the G-CSF + L + B group (I, n=12), and the G-CSF + L + Q + B (J, n=12). Mouse models of chemotherapy related leukocytopenia were established by intraperitoneal injection of cyclophosphamide (CTX). A G-CSF group was set up as a positive control. Mice were treated by a single oral drug, a single oral drug combined with G-CSF, and two or three drugs combined with G-CSF respectively, and the death rate calculated. Hemocytes [such as white blood cells (WBC) and its classification, red blood cells (RBC), platelet (PLT), hemoglobin (Hb)] were calculated by hematology analyzer. Mice were anatomized and important organs weighed. Organ indices were calculated.

RESULTSThere was no statistical difference in the mortality rate among all groups (P > 0.05). Compared with Group B, WBC was elevated in all other groups (P < 0.01). WBC and PLT were elevated most in Group J, Hb and RBC were also increased at the same time (P < 0.05, P < 0. 01). Compared with Group B, RBC increased in Group E, F, G, I, and J (P < 0.01); Hb obviously increased in Group C, E, F, H, I, and J (P<0.01). Compared with Group B and D, the promotion of erythroid hematopoiesis by G-CSF could be elevated in any group contained drug B and L (P < 0.05, P < 0.01). The spleen index of model mice could be significantly improved in Group C, D, and G (P < 0.01). The thymus index of model mice could be significantly improved in Group H (P < 0.05).

CONCLUSIONSThe best scheme to treat mice with chemotherapy related leukopenia or decreased three blood series was to administrate three commonly oral drugs combined with G-CSF. Authors speculated that G-CSF and Q might have a certain effect on CTX induced immune inhibition.

Administration, Oral ; Animals ; Blood Platelets ; Cyclophosphamide ; Drug-Related Side Effects and Adverse Reactions ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Erythrocyte Count ; Granulocyte Colony-Stimulating Factor ; metabolism ; Hematopoiesis ; Hemoglobins ; Leukocyte Count ; Leukocytes ; Leukopenia ; chemically induced ; drug therapy ; Male ; Mice ; Pharmaceutical Preparations

Objective To study the efficacy and mechanism of compound Tripterygium hypoglaucum Hutch (THH) on photoallergic contact dermatitis in mice. Methods The photoallergic animal model of BALB/c mice was established by using photosensitizer chlorpromazine and UVA irradiation. The therepeutic efficacy was determined by measuring the thickness and the weight of the swelling ear and the number of infiltrated mononuclear cells in the ear tissue. Immunohistochemical technique was used to detect the ICAM-1 expression on keratinocytes, fibroblasts and vascular endothelial cells. The serum level of INF-? was measured by ELISA. The tested animals were divided into 3 groups: compound THH, THH alone and normal saline. Results The difference of the thickness of left ear before and after challenge, the differences of the thickness and the weight of ear tissue, the difference of the number of infiltrated mononuclear cells of left and right ear after challenge were significantly less in the compound THH group than those in the THH alone group (P

Adenocarcinoma ; metabolism ; pathology ; Adenoma ; metabolism ; pathology ; Aged ; Cell Line, Tumor ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Ether-A-Go-Go Potassium Channels ; genetics ; metabolism ; Female ; HT29 Cells ; metabolism ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Tumor Burden

OBJECTIVETo observe the effect of pure total flavonoids from Citrus (PTFC) on the hepatic fatty degeneration, inflammation, oxidative stress and SIRT1/PGC-1alpha expressions in mice with non-alcohol steatohepatitis (NASH), and discuss the action mechanism of PTFC on NASH.

METHODMice were given high-fat diet for 16 weeks to induce the NASH model. Since the seventh week after the model establishment, the mice were intervened with 100, 50 and 25 mg x kg(-1) x d(-1) PTFC for 10 weeks. The pathologic changes in hepatic tissues were observed with HE staining. The contents of TG, CHOL in hepatic tissue, as well as the levels of AST, ALT in serum were detected by using the biochemical process. The expression of SIRT1, PGC-1alpha and MnSOD mRNA in hepatic tissues were detected with Real-time PCR assay. SIRT1, PGC-1alpha protein and 8-OHdG expressions were determined with the immunohistochemical method. The SOD level in hepatic tissues was tested by the xanthine oxidase method. The MDA content in hepatic tissues was examined by the thiobarbituric acid method.

RESULTThe contents of TG, CHOL, NAFLD activity scores and ALT level in serum in hepatic tissues of mice in the model induced by fat-rich diet were obviously higher than that of the normal group (P < 0.010. The SIRT1, PGC-1alpha, MnSOD mRNA and protein expression in hepatic tissues were significantly lower than that of the normal group (P < 0.01). The expression of 8-OHdG and the content of MDA in hepatic tissues were obviously higher than that of the normal group (P < 0.01). After the intervention with different doses of PTFC, the NAFLD activity scores, the content of TG and the level of AST in serum were notably lower than that of the normal group (P < 0.01, P < 0.05); whereas the SIRT1, PGC-1alpha, MnSOD mRNA and protein expression were obviously higher than that of the normal group (P < 0.01, P < 0.05), with the significant decrease in the expression of 8-OHdG and the content of MDA (P < 0.01).

CONCLUSIONOxidative stress/lipid peroxidation enhancement in in NASH mice induced by high-fat diet may be related to the changes in SIRT1/PGC-1alpha signal transduction pathway. PTFC could enhance the anti-oxidant capacity in liver, relieve the damage of reactive oxygen during the fatty acid metabolic process, and prevent NASH from the occurrence and development by regulating the SIRT1/PGC-1alpha signal pathway.

Animals ; Citrus ; chemistry ; Fatty Liver ; drug therapy ; genetics ; metabolism ; Flavonoids ; chemistry ; pharmacology ; Inflammation ; drug therapy ; genetics ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; Oxidative Stress ; drug effects ; genetics ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Sirtuin 1 ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism