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The scaffolding teaching is a kind of "student-centered" teaching desgin. In this study, the teacher breaks down complex content into simply conceptual framework so as to enable students to define their learning objectives, provide help for students' learning, and guide students to think independently and explore cooperatively to construct their own knowledge system. Clinical Microbiology Laboratory Technology is a specialized course with very complicated theoretical contents, which can break up the whole into parts by scaffolding instruction, combined with the learning strategies of cooperation, exploration and mind mapping. Finally, "installment fixed deposit" is realized by induction and summary. The scaffolding teaching model promotes students' ability of autonomous learning, analytical reasoning and innovation.
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We investigated the correlation between toxin gene exoS,exoU and fluoroquinolone resistance in lower respiratory tract infection with P.aeruginosa so as to provide guidance for reasonable treatment of clinical infections.We collected P.aeruginosa of sputum samples in hospitalized patients from October 2015 to March 2016.The antimicrobial susceptibility was tested by liquid dilution method.The exoS and exoU genes were detected by PCR technique.Results showed that forty-six P.aeruginosa strains were identified from sputum.The exoS and exoU gene positive rate were 86.96 ％ (40/46) and 69.57 ％ (32/ 46) respectively,and the highest proportion of genotype was exoS+/exoU+ (60.87％,28/46).Among them,36.96％ (17/ 46) were multiple drug-resistant bacteria(MDR).Fluoroquinolone non-sensitive (FQ-NS) strain were 78.95％ (15/19) for MDR and 89.47 ％ (17/19) exoU gene were positive,which was significantly higher than the fluoroquinolone sensitive strains (FQ-S).Compared with the FQ-S strain,FQ-NS strains were serious drug resistance.The drug resistant rate of eefepime and aztreonam were more than 70％,and then meropenem and imipenem were more than 50％.The drugs of lower resistance rate in FQ-NS strain had polymyxin B(10.53％,2/19),amikacin(10.53％,2/19),ceftazidime (15.79％,3/19) and gentamicin (21.05％,4/19).P.aeruginosa of lower respiratory infection carried toxin genes exoS and exoU were higher,the main genetpy was exoS+/exoU+.FQ-NS strains were higher drug resistance rate and a higher proportion of exoU+ strains than FQ-S strains.We should strengthen virulence genes test and drug resistance monitoring in clinical practice.
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Objective:To explore the effects of voriconazole on the proliferation and morphology of Acanthamoeba cultivated in vitro,and to clarify the killing effects of voriconazole against the trophozoites and cysts of Acanthamoeba.Methods:The Acanthamoeba polyphaga at logarithmic phase were selected and divided into control group and experiment groups(2.5 and 25.0 mg·L-1).The Acanthamoeba in each group was collected at 24,48,72,and 96 h after drug administration,respectively.Then the concentrations of Acanthamoeba were calculated and the proliferation curves were drawn;inverted microscope was used to observe the morphology,activity and adherence of Acanthamoeba;the ultrastructures of Acanthamoeba were observed under electron microscope.Results:Compared with control group,the numbers of Acanthamoeba polyphaga in experiment groups were significantly decreased(P<0.01).The morphology of Acanthamoeba changed significantly under inverted microscope,and the shape of Acanthamoeba transformed from the trophozoites with irregular spiny filopodia to circular cysts.Even a large number of cell debris was observed.Different degrees of damage and even necrosis of Acanthamoeba in experiment groups were found under electron micro scope.Conclusion:Certain concentration of voriconazole can effectively inhibit the proliferation of Acanthamoeba and change the morphology and ultrastructure and kill the trophozoites and cysts of Acanthamoeba cultivated in vitro.
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Objective To investigate the expression of anoctamin 1 in Chinese hamster ovary cells (CHO)and to analyze the functional properties of its ion channel,and to provide experimental basis for study on the physiological function of calcium-activated chloride channel.Methods The whole sequence of anoctamin 1 was obtained by PCR technique and was subcloned into pcDNA3.1 to construct the expression vector pcDNA3.1-anoctamin 1 was transfected into CHO by liposome-mediated transfection and the CHO stably expressing anoctamin 1 were aquired by selection with zeocin. The expression and distribution of anoctamin 1 in CHO were measured by RT-PCR technique and inverted fluorescence microscope.The functional properties of anoctamin 1 were measured with halide-sensitive fluorescence proteins YFP-H148Q/I152L.The PBS buffer solution with calcimycin and high concentration of iodine ion was used as experimental group,andthe PBS buffer solution without calcimycin and high concentration of iodine ion was used as control group.Results The results of digestion and sequencing confirmed that anoctamin 1 was cloned into pcDNA3.1 by electrophoresis and blast. The results of RT-PCR and inverted fluorescence microscope indicated that anoctamin 1 was expressed in CHO. The results of I- influx as measured by halide-sensitive fluorescence proteins YFP-H148Q/I152L showed that anoctamin 1 had the more functional properties of trans-epithelial transporting I-,and the transporting speed in experimental group was higher than that in control group (P<0.05).Conclusion Anoctamin 1 can be expressed highly in the CHO;Anoctamin 1 expressed in CHO has the characteristics of calcium-activated chloride channel.
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AIM:To investigate the expression of transmembrane protein 16A(TMEM16A) in Fischer rat thy-roid follicular epithelial ( FRT) cells and its electrophysiologic properties .METHODS: The eukaryotic expression vector of pUB6/V5-TMEM16A was constructed and transfected into FRT cells by liposome-mediated transfection .In order to ob-tain the high efficiency of gene transfection and expression , the quantity and ratio of lipid/DNA complexes were optimized . The FRT cells stably expressing TMEM16A were gained by the selection with blasticidin and confirmed by the techniques of RT-PCR and immunofluorescence .The expression and location of TMEM 16A in the FRT cells were observed under an in-verted fluorescence microscope .TMEM16A protein was associated with calcium-dependent chloride current , as measured with halide-sensitive fluorescent protein and patch-clamp technique .RESULTS: The results of double digestion and se-quencing indicated that TMEM16A was cloned into pUB6/V5.The results of RT-PCR and immunofluorescence confirmed that TMEM16A was expressed in the FRT cells after transfection with TMEM16A.The classical calcium-activated chloride channel currents were recorded in the FRT cells stably expressing TMEM 16A by the technique of patch-clamp and halide-sensitive fluorescent protein YFP-H148Q/I152L.CONCLUSION:The protein expression of TMEM16A in the FRT cells was observed.TMEM16A is the molecular identity of calcium-activated chloride channels .
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BACKGROUND: Vitrification is a comparatively new technology which applies high concentration cryoprotectant and rapid refrigeration. By the method, the cells were quickly frozen and to avoid damage by ice crystals inside and outside. OBJECTIVE: To compare the effect of four cryoprotectants on morphology and function of ovarian tissue in rats after vitrification. METHODS: The rats were randomly assigned into six groups with 6 rats for each: DMSO + EG, DMSO + EG + sucrose, DMSC +EG + sucrose + acetamide, EG + sucrose + acetamide, ovariectomized, and normal control groups. The ovarian tissues of four freezing groups were treated with the corresponding cryoprotectants, the vitrified ovarian tissues were then resected but not frozen and transplanted; otherwise, tissues were not treated with any treatment in the normal control group. Two weeks after freezing, the tissues were thawed and heterotopic-transplanted into femoribus intemus of hind limb. At 30 days after implantation, vaginal epithelial cells and estrus cycle were observed, while after three months, blood were collected to detect the level of estradiol (E2) and the ovarian tissues were reclaimed to analyze their morphological changes. RESULTS AND CONCLUSION: All ovarian tissues were damaged after cryoprersarvation in four freezing groups. The rates ot healthy primordial follicles were 67.9%, 71.6%, 80.5%, and 59.4%, respectively, while healthy primary follicles were 41.6%, 52.3%, 55.9%, and 36.7%, respectively. In all freezing groups, the rate of the healthy follicles in DMSO + EG + sucrose + acetamide group was higher than DMSO + EG group and EG + sucrose + acetamide group (P < 0.05). No significant difference was found in the proportion of follicles at different development stages among four groups. The typical secondary follicle was not found in four groups. Damaged ovotid showed oocyte pyknosis and vacuolation in cytoplasmic area. There was not typical cell type of all freezing groups. Ovarian autografting gained visible vascularity from surrounding tissue that connected ovarian tissue to form net. There was a lot of blood capillary in transplanted ovarian tissues and clumped primordial follicles in cortical substance. The rates of primary follicles and secondary follicles were lower than primordial follicles. The level of serum estradiol was obviously decreased compared with normal control group (P < 0.01). There was significant difference between DMSO + EG + sucrose + acetamide group and other three freezing groups (P < 0.05). Four kinds of freezing methods have poor effects on different stages of follicles and the structure of ovariarn tissue. DMSO + EG + sucrose + acetamide group is an optimal protocol for cryoprerserving ovarian tissue. Freezing methods still need to explore further because the rats had not appeared disciplinary estrus cycle after ovarian autoqrafting.
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In recent years, with the development of neuroimaging and the advancement in related studies, people have had a deeper understanding of perimesencephalic nonaneurysmal subarachnoid hemorrhage. This article reviews the progress in research on the etiology, onset characteristics, imaging features, diagnosis and prognosis of perimesencephalic nonaneurysmal subarachnoid hemorrhage.
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AIM: To study the effect of the radix sophorae flavescentis on cellular immunity in rats with Pneumocystis Carinii Pneumonia (PCP) induced by long-term use of immunosuppressant, and explore the action of traditional Chinese medicine for the immunological regulation and infectious prevention after organ transplantation. METHODS: The experiment was conducted at Department of Pathobiology, Jilin Medical College from May 2005 to March 2006. Forty adult healthy female SD rats were selected from Harbin Medical University (Certification: 02473146) and randomly divided into experiment group and control group, with 20 rats in each. The model of PCP was set up by glucocorticoid injection subcutaneously to SD rats (25 mg once, 2 times/week). The mixture of sophorae flavescentis was given to stomach with tube in experiment group (3 mL/kg, 2 times/day), and was consisted of radix sophorae flavescentis, ash bark, amur cork-tree, malt, milkvetch root and danshen root. Six weeks later, all the rats were anesthetized and broncholveolar lavage fluids were collected.①Alveolar washing fluid was concentrated 10 times and the levels of the soluble interleukin-2 receptor (sIL-2R) were examined by double antigen sandwich ELISA.②Blood was sampled from rat eyes and the count of lymphocytes in peripheral blood were detected.③The percentages of CD3+ and CD4+ subgroups were assessed with erythrocyte chaplet kit sensitized by antigen. RESULTS: All 40 rats were involved in the result analysis without drop.①The count of lymphocytes in peripheral blood in experiment group was significantly higher than that in control group (5.1?1.3)%, (0.8?0.3)%, P
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The experiment was performed at the Institute of Treatment and Prevention of Tumor of Jilin Province and Department of Etiology, Jilin Medical College from October 2004 to June 2005. The normal umbilical cord blood (UCB) was provided by a healthy lying-in woman from Changchun Hospital with the permission of the pregnant woman. Full-term normal delivery and UCB of healthy pregnant women was collected by aseptic venepuncture after fetal disengagement, and with Natrium Citricum anticoagulation. After the UCB mononuclear cells were isolated by Ficoll-paque (relative volume mass: 1.077?0.22) density gradient centrifugation, long-term liquid culture system was used for UCB adherent cells, and co-cultured with UCB mononuclear cells. The adherent cells were observed and mononuclear cell cycles were tested with flow cytometer. It showed that adherent layer of UCB increased UCB mononuclear cells expansion as compared with that of non-adherent layer of UCB after co-culture for 7 days. The cell percentage in S+G2+M phase obviously increased, and there was significant difference [(42.7?1.1)%,(35.5?2.8)%,P
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Objective To identify the changes in proliferative activity and cytotoxicity of lymphokine activated killer(LAK) cells after in vitro co-culture with dendritic cells from umbilical cord blood(CBDC). Methods CBDCs were induced by using GM-CSF and IL-4,and then co-cultrued with allogenic LAK at different proportions.Proliferative activity and cytotoxicity of LAK cells were measured by MTT assay.Results The typical CBDCs were induced in vitro,and the cultured CBDCs stimulated the proliferation and cytotoxicity of LAK cells.The optimal proportion of CBDC and LAK was 1:10,at which the activated LAK showed the highest cytotoxic effect on the tumor cells.Conclusion The CBDCs can enhance the proliferation and cytotoxicity of LAK cells.