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Objective:To explore applications of essential balm K-point stimulation on the swallowing rehabilitation in patients with minimally consciousness state, so as to provide reference for patients rehabilitation.Methods:This was a prospective randomized controlled study. A total of 60 minimally consciousness state patients were enrolled by convenience sampling method from Henan Provincial People Hospital from January to December 2021, who were divided into experimental group and control group according to the random number table method, with 30 cases in each group. Both groups received routine rehabilitation therapy, based on this, the control group was given routine K-point stimulation, the experimental group implemented essential balm K-point stimulation. Before and after 4 weeks of intervention, the Kubota water swallowing test and Coma Recovery Scale-revised (CRS-R) was used for assessment.Results:Before intervention, there was no significant difference in Kubota water swallowing test grade and CRS-R scores between two groups (both P>0.05). After 4-week intervention, Kubota water swallowing test Ⅰ-Ⅴ grade were 1, 6, 13, 7 and 3 cases in the control group, while 4, 12, 8, 3 and 3 cases in the experimental group, the difference was statistically significant ( Z = 2.09, P<0.05); the scores of oral motor/verbal function scale were (2.60 ± 1.16) in the experimental group, higher than in the control group (1.90 ± 0.32), the difference was statistically significant ( t = 2.18, P<0.05). Conclusions:Essential balm K-point stimulation can effectively promote swallowing function of patients with minimally consciousness state, and improve patient′s oral motor function.
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Objective To explore the method and effect of applying just-in-time-teaching (JiTT) mode in the reform of the teaching method of biochemical comprehensive experiment (Isolation, Purification and Identification of γ-Globulin From serum). Methods A total of 100 undergraduate students in our 2016 clinical medicine were divided evenly into two groups: the control group and the experimental group. The control group used traditional teaching, and the experimental group used JiTT-based teaching. In the experimental group, an online environment for pre-classroom discussion and feedback was constructed by uploading learning resources such as short video and tutorials to the online teaching platform and the online learning exchange group 3-5 days before the class; the class session itself consisted of personal tests, face-to-face teaching by teachers, group discussion, finishing experimental report and answering questions etc., and was intended to evaluate the students' self-study. SPSS 22.0 was used to collect exam scores and questionnaire results, and the data between groups were compared using t test. Results The scores of individual test scores in the experimental group (9.44±0.59) were higher than those in the control group (8.77±0.41) (P=0.00), and the difference was statistically significant. In all the six questionnaire items pertaining to teaching effect , JiTT teaching method received higher scores than the traditional teaching method (P<0.05), and the difference was statistically significant. Conclusion JiTT can improve the quality of biochemistry experiment teaching. It is worthwhile to try it in the teaching of other basic specialized courses.
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Objective To investigate the endogenous carnosine,glutathione system and oxidative stress level in serum of patients with diabetic cardiomyopathy(DCM),and the potential relationships among them.Meth-ods The serum of 102 healthy people,96 patients with type 2 diabetes mellitus and 74 patients with diabetic car-diomyopathy in the third affiliated hospital of QiQihar Medical University were enrolled.Carnosine content was mea-sured by ELISA.T-GSH,GSH and GSSG were detected by using micro-enzyme labeling assay.GSH-Px,GST and GRAC were detected by using colorimetry. The content of NO was detected by nitrate reduction method. The con-tent of H2O2was detected by using molybdic acid coloring method. The activities of NOS and CAT was determined by colorimetry. Results Compared with the healthy control group,the average level of human serum carnosine, GSH content,GRAC,GSH/GSSG ratio,GST activity and T-SH content in the DM and DCM group were signifi-cantly reduced(P < 0.05,respectively). NO content,H2O2and iNOS activity were increased,but CAT activity was decreased in DCM group.Conclusion Decreases of levels of serum carnosine and glutathione(GSH)and the imbalance of redox state were observed in patients with diabetic cardiomyopathy,which may promote the occur-rence and development of the diabetic cardiomyopathy.
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Objective To discuss the effect of the teaching mode of technological pedagogical and content knowledge in biochemistry theory teaching. Methods 400 students in 8 classes of clinical medicine undergraduate in Grade 2013 were selected and randomly divided into 2 groups. The traditional teaching mode was applied in the control group of 198 students in 1-4 classes, while the technological pedagogical and content knowledge teaching mode was applied in the experimental group of 202 students in 5-8 classes. 385 students in 8 classes of clinical medical undergraduates in Grade 2014 were selected and randomly divided into 2 groups. The control group of 189 students in 1-4 classes adopted the traditional teaching mode, while the experimental group of 196 students in 5-8 classes adopted the technological pedagogical and content knowledge teaching mode, which used the micro-lesson and network platform as learning resource superior to the flipped classroom. Flipped classroom was divided into two major learning links:extracurricular self-study and class digestion. Through the network platform the micro-lesson was presented to the learners. Learners could make self-study according to their own specific circumstances and in the classroom many activities were increased such as the mutual cooperation between the students, the students' PPT teaching, the students' questions and the discussion, and the interaction between teachers and students, etc. The results of the examinations of the two terms students of the same profession and the questionnaire were analyzed. The related data were processed by SPSS 15.0, and the data between groups were compared by t test . Results The test scores analysis showed that the individual test scores in experimental group of Grade 2013 were [(17.94±2.02) vs. (12.28±4.17)], and the individual test scores in experimental group of Grade 2014 were [(18.21 ±1.78) vs. (12.45 ±5.13)], which were obviously higher than the control group, and there was statistical significance. The final exam scores in experimental group of Grade 2013 were [(78.28±11.18) vs. (68.65±12.51)], and the final exam scores in experimental group of Grade 2014 were [(81.73 ±9.12) vs. (74.41 ±11.87)], which were obviously higher than the control group, and there was statistical significance. The results of survey showed that the students thought the teaching mode aroused their study interests while 393 (93.7%), thought the teaching mode developed their self-study ability while 357 (89.7%), thought the teaching mode beneficial to cultivating their ability of solving the problems. Conclusion The teaching mode of technological pedagogical and content knowledge is of certain signifi-cance to break the plight of the traditional teaching, inspire the students interest in learning, improve the teaching quality of biochemistry, and make for the teachers' professional development.
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Trace elements is the main content in Biochemistry and Molecular Biology courses through students' self-study.This topic takes the trace elements as the content of open video courses to make the medical students and the public be able to scientifically understand the relationship between trace elements and human health and pay attention to the influence of dietary nutrition and environmental pollution on human society.This course is based on audience online teaching rules and online learning needs,to achieve high-quality resources sharing and popularity as the goal,using typical case,vividly describes the source of the common trace element,function,and the relationship between health and disease.In the course construction,we realize that the selection of teaching content,optimization of teachers,professional recording team are the basic elements of the success of open video courses production.The combination of open video courses and flipped classroom teaching has gained a high degree of satisfaction in improving students' independent learning ability,expanding knowledge and so on.
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AIM:To investigate the effect of the N-terminal 24-amino acid (N24) overexpression in p55γre-gulatory subunit of phosphatidylinositiol 3-kinase ( PI3K) on the cell adhesion of human gastric carcinoma cell MGC 803. METHODS:MGC803 cells, which stably expressed GFP-N24 fusion protein and GFP alone , were generated by transfec-tion with pEGFPN-24 plasmid and control plasmid pEGFP-C1, respectively.The morphological change of the cells was ob-served under inverted microscope .The expression of GFP-N24 fusion protein was detected by Western blot .The adhesion of the cells was determined by cell adhesion assay .The effects of GFP-N24 fusion protein on the expression of E-cadherin andβ-catenin were analyzed by Western blot .The expression and secretion of matrix metalloproteinase 9 (MMP9) and u-rokinase-type plasminogen activator ( uPA ) in the MGC803 cells were detected by gelatin zymography .RESULTS:MGC803/GFP-N24 cell line steadily expressed GFP-N24 fusion protein and MGC803/GFP cell line steadily expressing GFP were successfully established , but the expression of fusion protein GFP-N24 was lower than that of the control protein GFP.The morphological changes of the transfected cells from paving stone to fibroblast cell form after gene transfection , and the cytoplasm secretory granules were increased significantly .The cell adhesion to fibronectin and collagen decreased after GFP-N24 transfection .GFP-N24 fusion protein increased the expression of cell adhesion molecule E-cadherin and de-creased the wnt signal pathway molecule β-catenin in the MGC803 cells.However, it did not affect the expression and se-cretion of tumor metastasis-related proteins MMP9 and uPA.CONCLUSION:Overexpression of N24p55γinhibits cell ad-hesion by influencing the expression of E-cadherin and β-catenin in the MGC803 cells.
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Objective Analysis the expression and significance of P16INK4 ,P57kip2 and Ki67 in hepatocellular carcinoma .Meth-ods 21 normal liver tissue ,48 hepatocellular carcinoma tissues and 48 corresponding adjacent tissue were collected for this study . Immunohistochemical analysis the expression levels of P16INK4 ,P57kip2 and Ki67 .Results P16INK4 and P57kip2 in hepatocellular carci-noma tissue were significantly lower ,the positive rates were 49 .3% and 34 .2% respectively ,Ki67 was significantly higher than the normal liver tissue and corresponding adjacent tissue ,the difference was statistically significant(P<0 .05) ,the results were consist-ent with the Western blot ;different differentiated group p57kip2 ,Ki67 expression and P16INK4 were significant differences(P<0 .05) . Conclusion Low expression of P16INK4 and P57kip2 ,the higher expression of Ki67 play an important role in the development of hepa-tocellular carcinoma .
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Objective To explore teaching effectsof biochemistry by comparingthe traditional teaching with the network investigative teaching. Methods 425 five-year-program students of clinical medicine of grade 2012 were chosen as subjects, with 246 students from Classes 5-8 as experimental group in which network investigative teaching was implemented andwith 179 students of Classes 17-20 as the control group which wasgiven traditional teaching. After the teaching , the teaching effects were evaluated through a questionnaire and the test scores of the two groups were compared and statistically analyzed. The two groups were compared by using t test. Results More than 70% of students in the experimental groups were generally satisfied with several aspects of the network investigative teaching and gavea good evaluation. The experimental group students' total scores and every item scores were all higher than the control group students', in which the practise scores and the case analysis problem scores had significant differences by statistical analysis with P values being 0.000 and 0.002, respec-tively. Conclusion Network investigative teaching is better than traditional teaching in the teaching of biochemistry , which can enhance students' ability of problem analysis and their enthusiasm for learning.
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<p><b>OBJECTIVE</b>To evaluate and explore the effects of 20(S)-ginsenoside Rh2 and 20(R)-ginsenoside Rh2 on the cytotoxicity, proliferation and the apoptosis of human lung adenocarcinoma A549 cells, and to illustrate the structure-activity relationship and possible mechanisms of anti-tumor active ingredients of ginseng.</p><p><b>METHOD</b>A549 cells were treated with different concentration gradient of ginsenoside Rh2 (S and R structure) and incubated for different time. Cell proliferation and cytotoxicity studies were detected by methyl thiazolyl tetrazolium (MTT) colorimetric assay, cell cycle and apoptotic was analyzed by PI stains and combination of Annexin V/Prop idium iodide double staining with flow cytometric analysis. The influences of activation on Caspase-3 were also detected by the immunofluorescence staining with fluorescence microscope.</p><p><b>RESULT</b>MTT test indicated that ginsenoside Rh2 had a strong cytotoxicity activity to A549 cells. Ginsenoside Rh2 could obviously inhibit the cell proliferation in human lung adenocarcinoma cell line A549 at the effective doses of 25 mg x L(-1) treated with 48 h. The inhibition ratio and the value of IC50 for48 h of 20(R)-Rh2 and 20(S)-Rh2 were respectively 28.5%, 33.6% and 33.4, 28.5 mg x L(-1). The inhibition of ginsenoside Rh2 to A549 showed structure relationship significantly, time-dependent and concentration-dependent. Flow cytometric analysis (FACS) with PI stains analysis results showed that the proportion of A549 cells in G1 phase increased, while the number of cells in S phase decreased significantly and those in G2 phase reduced slightly. This result indicated structure relationship significantly, especially in the 20(S) -ginsenoside Rh2 inhibited the proliferation of A549 cell dramatically and retarded A549 cell cycle at G0/G1 phase. The immunofluorescent of combination with Annexin VFITC/PI by flow cytometric suggested ginsenoside Rh2 can induce inchoate apoptsis rate and late apoptosis rate of A549 cell significantly. All the results showed structure relationship significantly, especially in the 20(S)-ginsenoside Rh2. The immunofluorescent with fluorescence microscope suggested the activity of Caspase-3 were enhanced after ginsenoside Rh2 treated.</p><p><b>CONCLUSION</b>20 (R) and 20(S)-ginsenoside Rh2 had a significant inhibitory effect on the proliferation. Compared with 20(S)-ginsenoside Rh2, 20 (S)-ginsenoside Rh2 has been shown to have significant anticancer effects and to be capable of blocking cell proliferation and causing G1 phase arrest in human lung adenocarcinoma A549 cells. 20 (R) and 20(S)-ginsenoside Rh2 have been shown to have anticancer effects and to be capable of increasing inchoate apoptotic rate, reducing apoptotic rate significantly, enhancing the activity of Caspase-3 and inducing apoptosis in human lung adenocarcinoma A549 cells.</p>
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Humanos , Apoptosis , Ciclo Celular , Línea Celular Tumoral , Proliferación Celular , Ginsenósidos , Farmacología , Microscopía FluorescenteRESUMEN
Objective To observe how geniposide as an anti-inflammatory agent through inhibition Toll-like receptor 7/nuclear factor-κB signaling pathways activation, as well as TNF-α and IL-6 release infectioned by influenza virus. Methods Epithelial cells was exposed to human influenza viruses A/Gui/81/23(H3N2) infection for 2 h before treatment with geniposide for 24 h. NF-κB responsive element luciferase reportor gene was transfected and dual luciferase cis-reporting systems was used to assay the transcriptional activity of NF-κB under the stimulated circumstance of influenza virus infection. The phosphory level and nuclear transposition of NF-κB was observed by fluorescence inverted microscope. RT-PCR was used to detect the gene transcription level of TLR7, TNF-α and IL-6. Results The relative luciferase reporter assay of NF-κB was apparently improved by influenza virus infection. But geniposide significantly repressed the relative value of luciferase. The phosphorylation level and rate for nuclear transposition of NF-κB was apparently improved by influenza A virus infection observed by fluorescence inverted microscope. But geniposide significantly repressed the phosphorylation level and rate for nuclear transposition. RT-PCR showed upregulation of TLR7 and pre-inflammatory markers TNF-α and IL-6 in A549 cells infected by influenza virus, geniposide had a significant effect on the expression of TLR7 and inflammatory markers TNF-α and IL-6 after treated with influenza virus. Conclusion Geniposide as an antiinflammatory agent antagonized influenza A virus infection through inhibiting Toll-like receptor 7/nuclear factor-κB signaling pathways activation, as well as on the downregulation of the downstream inflammatory markers target gene expression TNF-α and IL-6.
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Objective:To observe the influence of ginsenoside Rg1 on transcriptional activation of NF-κB induced by hydrogen peroxide (H_2O_2) in 293T cell,and probe into the antioxidant mechanism of ginsenoside Rg1.Methods:In the experiment,cells was exposed to H_2O_2 after pretreatment with Rg1,cell proliferation and cytotoxicity studies were detected by MTT and Trypan blue.The quantities of generation of intracellular reactive oxygen species (iROS) was analyzed by flow cytometric analysis measured with fluorescent probe 2,7-dichlorofluorescin diacetate (DCFH-DA).NF-κB-responsive element-luciferase reporter gene was transfected and dual-luciferase cis-reporting systems were used to assay the transcriptional activity of NF-κB under the stimulated circumstance of oxidative stress induced by H_2O_2.Results:The results of MTT showed that ginsenoside Rg1 apparently protected the proliferation of 293T cell,which were repressed by H_2O_2 (P＜0.05).The results by trypan blue showed that H_2O_2 stimulated substantial cytotoxicity.This effect was markedly attenuated by treatment with ginsenoside Rg1.Oxidant production,measured as the fluorescence of dichlorofluorescein,was significant increased by 40%-50% through H_2O_2 stimulation.The decrease in iROS generation was significant blocked by 35%-40% through Rg1 and antioxidant.The relative luciferase reporter assay of NF-κB was apparently improved by H_2O_2-induced(P＜0.05),but Ginsenoside Rg1 significantly repressed the relative value of luciferase (P＜0.05).Conclusion:Ginsenoside Rg1 has the obvious protective function from the damage of oxidative stress damage,whose possible mechanism is to eliminate excessive free radicals of the cells effectively,to reduce transcriptional activation of nuclear factor kappa B(NF-κB),and subsequently to suppress the NF-κB circuit activation.
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Objective To investigate the inhibitory effects of CIK on the proliferation of Lewis lung carcinoma cell line and the growth of transplanted Lewis lung carcinoma in C57BL/6N mice in vivo.Methods CIK cells were induced by culturing PBMC with regular method.The proliferation of Lewis lung carcinoma cells was measured by MTT assay.Uhramicrostrueture of Lewis lung carcinoma cells was observed under a transmission electron microscope.Flowcytometric analysis was used to detect cell apoptosis.Ultrastructural observation expressions of FasL were individually determined by MTT and immunocytochemistry(ICC) analysis.Results Electron microscopic observations sbowed that CIK cells could induce the hepatoma cells to apoptosis.Flow cytometric analysis demonstrated that apoptosis cells of Lewis lung carcinoma were increased in CIK group compared with those in the control group.FasL expression on CIK increased.Cytotoxieity was blocked after addition of anti-FasmAb.Conclusion CIK has inhibitory effect on Lewis lung carcinoma cells both in vitro and in vivo.CIK cells can induce the apoptosis of Lewis lung carcinoraa cells.and Fas/FasL pathway plays an important role in apoptosis of Lewis lung carcinoma cells by CIK.
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Objective To investigate the inhibitory effects of CIK cell on apoptosis of lung cancer cell A549 in vivo. Methods CIK cells were induced by culturing PBMC with regular method and cell apoptosis was detected.Results Electron microscopic observations showed that CIK cells could induce the lung cancer cells A549 to apoptosis. Flow cytometry(FCM)demonstrated that apoptosis cells of lung cancer cells A549 were increased in CIK group as compared with the control group. Conclution CIK cells can induce the apoptosis of lung cancer cells A549.
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Objective To investigate the role of glutaredoxin1(Grx1)on high glucose-induced apoptosis in cultured umbilical vein endothelial cells.Methods Human umbilical vein endothelial cells was cultured and induced with different dose of glucose and Grx1.We divided the cells into three groups:cell control group,damage group(high glucose group),pretreatment with Grx1 group(Grx1+ high glucose group).The morphological changes of the cells were observed by light microscope.The proliferation of cell was measured by MTT assay.The morphon of cell nucleolus of endothelial cell was observed in a fluorescence microscope by Hoechst 33258 stain and the influences of Grx1 on the apoptosis were determined by the immunofluorescent of Annexin V-FITC/PI with flow cytometer.Results Under the light microscope Grx1 ameliorated cells condition and restore the structure of organelle compared with damage group.Grx1 prevented the inhibitory effect on cell viability induced by high glucose;Hoechst33258 stains suggested Grx1 protect the cells nucleolus against high glucose-induced apoptosis.The analysis of Grx1 can restrain apoptosis rate of endothelial cell significantly.Conclusion Grx1 can obviously protect human umbilicus vein endothelial cells from apoptosis damages induced by high glucose.
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Objective To study the effects of ganoderma applanatum polysaccharides(GAPS) on cell morphology, proliferation and PDGFR -?expression in cell lines MGC - 803 , and to explore its potential mechanisms of anti - tumor of GAPS. Methods Cell morphology was observed by inverting microscope. MTT assay was used to investigate the inhibitory effect of GAPS on MGC -803.Expressions of PDGFR -?was analyzed by flow cytometry (FCW). The fluorescence intensity of expressions of PDGFR -?was observed by fluorescence microscope. Results Cells of GAPS group were irregular shaped and grew poorly. GAPS inhibited the proliferation of MGC -803 cells in dose - dependent and time - dependent manners . After MGC - 803 cells were treated with GAPS for 72h, GAPS could down - regulate expression of PDGFR -?observed by fluorescence microscope which was consistented with the results of FCW with statistic significance difference as compared with control group (P
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0.05). Compared with control cells, the relative luciferase activity of NF-?B in virus-infected cells was apparently up-regulated (P