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With the rapid development of information technology and the change in military education policy in the new era, online teaching has gradually become one of the main approaches to implement the clinical medical education in military teaching hospitals. In this study, the online teaching was performed mainly by pre-recorded teaching, supplemented by online live teaching. Massive open online courses (MOOCs) and the seminar-style teaching were advocated to be used in advanced disciplines. The quality of online teaching was guaranteed through infrastructure provision, teacher arrangement, teaching preparation, teaching interactivity, after-class test, teaching evaluation, supervision, and summary of teaching, which ultimately achieved a good effect. The results of the two-way questionnaires of 26 teachers and 129 students showed that 23.26% (30/129) of students and 65.38% (17/26) of teachers believed that students' learning ability was insufficient. 34.88% (45/129) of students and 23.08% (6/26) of teachers thought that the existing technology could not meet the requirements of online teaching. 55.04% (71/129) of students and 69.23% (18/26) of teachers held the view that the effect of online teaching was inferior to face-to-face teaching. 28.68% (37/129) of students and 57.69% (15/26) of faculty asked for the return of face-to-face education. Therefore, it is necessary to strengthen the construction of smart classroom platforms, establish a well-developed online teaching quality evaluation system, and integrate various innovative teaching modes with the online teaching. In these ways, it is expected to optimize the online teaching of clinical medicine and achieve the goal of online teaching.
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Objective To understand the frequency distribution of CYP2C9 and VKORC1 gene single nucleotide polymor-phisms in Yunnan Han population.Methods CYP2C9(430C> T,1075A>C and 1080C> G)locus and VKORC1(-1639G> A and 1173C> T)locus gene polymorphisms in 202 samples were detected by adopting electrochemical gene sensor method,and the allele frequencies and genotype frequencies were performed the statistics and the gene polymorphism in relevant populations was an-alyzed.Results Among 202 samples,202 cases(100.0%)were genotype C/C at CYP2C9 * 2 locus,C allele frequency was 100.0%;185 cases(91.6%)were genotype A/A at CYP2C9*3 locus,15 cases(7.4%)were A/C genotype,2 cases(1.0%)were C/C genotype,A allele frequency was 95.3%,C allele frequency was 4.7%;CYP2C9*5 locus genotype C/C was in 202 cases (100.0%),C allele frequency was 100%;VKORC1 -1639G > A locus genotype A/A was in 145 cases(71.8%),57 cases (28.2%)were G/A genotype,A allele frequency was 85.9%,G was 14.1%;1173C> T locus genotype T/T was in 145 cases (71.8%),C/T gene type in 57 cases(28.2%),T allele frequency was 85.9%,and C was 14.1%.Conclusion The distribution of CYP2C9 gene in Yunnan Han population is similar to that in other regions of our country.The VKORC1 gene is different from the foreign population,Chinese Han nationality and partial minority nationalities.
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Objective@#To explore the effects of hypoxia inducible factor-1α (HIF-1α) on P311 and its influence on the migration of murine epidermal stem cells (ESCs) under hypoxia in vitro.@*Methods@#Two kinds of murine ESCs were isolated and obtained from 15 neonatal wild-type C57BL/6J mice and 5 congeneric source P311 gene knock-out mice, respectively. The first passage of cells were used in the following experiments after morphologic observation and detection of expression of cell surface markers CD71 and CD49f with flow cytometer. (1) After cell scratch assay, according to the random number table (the same dividing method below), ESCs of P311 gene knock-out mice were divided into normoxia group (cells were cultured with complete medium in normoxic carbon dioxide incubator, and the subsequent normoxic treatments were the same) and hypoxia group (cells were cultured in hypoxic carbon dioxide incubator containing 1% oxygen, and the subsequent hypoxic treatments were the same), with 12 inserts in each group. ESCs of wild-type mice were divided into normoxia group, pure hypoxia group, dimethyl sulfoxide (DMSO) control group (2 μL DMSO solvent was added for 1 h of normoxia treatment before hypoxia treatment), HIF-1α inhibitor group (cells were treated with 11 μmol/L HIF-1 inhibitor of 2 μL under normoxia condition for 1 h before hypoxia treatment), HIF-1α stabilizer group (the cells were treated with 2 μmol/L FG-4592 of 2 μL under normoxia condition for 1 h before hypoxia treatment), with 12 inserts in each group. Three inserts of each time point in each group were adopted respectively to measure the residual width of scratch under inverted phase contrast microscope at post scratch hour (PSH) 0 (immediately), 12, 24, and 48. (2) After hypoxia treatment, the protein level of HIF-1α in ESCs of wild-type mice was detected by Western blotting at post hypoxia hour (PHH) 0, 12, 24, and 48. (3) ESCs of wild-type mice were divided into pure hypoxia group, DMSO control group, HIF-1α inhibitor group, and HIF-1α stabilizer group as that of experiment (1) with the same treatment. The mRNA expression of P311 and expression of P311 in ESCs were determined by real-time fluorescent quantitative reverse transcription polymerase chain reaction and immunocytochemical staining, respectively, at PHH 0 (immediately), 12, 24, and 48 (with sample numbers of 12). (4) The second passage of human embryonic kidney 293 (HEK-293) cells were divided into empty vector hypoxia group (cells were cultured under hypoxia condition after being transfected with empty vector plasmid), P311 normoxia group (cells were cultured under normoxia condition after being transfected with P311 reporter gene plasmid), P311 hypoxia group (cells were cultured under hypoxia condition after being transfected with P311 reporter gene plasmid), P311 hypoxia+ HIF-1α inhibitor group (cells which were incubated with HIF-1α inhibitor were cultured under hypoxia condition after being transfected with P311 reporter gene plasmid). The luciferase activity was detected at post culture hour (PCH) 0 and 12, respectively, and then the P311 transcriptional regulatory binding site of HIF-1α and the promoter sequence of P311 were predicted and searched by bioinformatics methods. Data were processed with factorial design variance analysis, one-way analysis of variance, LSD test and Bonferroni correction.@*Results@#(1) The results of ESCs. The cells showed cobblestone-like pattern and different clonal morphology due to the different cell proliferation potential. The proportion of CD71-CD49f+ cells accounted for about 85%. The identification results indicated that the cells showed strong stem cell properties and high purity. Compared with those in cells of normoxia group of P311 gene knock-out mice, the residual widths of scratch of cells in pure hypoxia group were smaller at PSH 12 and 24 (with P values below 0.05), and those in hypoxia group, normoxia group of wild-type mice, DMSO control group, HIF-1α inhibitor group, and HIF-1α stabilizer group were smaller at PSH 12 (with P values below 0.05). Compared with those in cells of normoxia group of wild-type mice, the residual widths of scratch of cells in hypoxia group, pure hypoxia group, and DMSO control group were smaller at PSH 12 and 24 (with P values below 0.05), and the residual width of scratch of cells in HIF-1α stabilizer group was smaller at PSH 12 (P<0.05). Compared with those of cells in pure hypoxia group, the residual widths of scratch of cells in hypoxia group were wider at PSH 12 and 24 (with P values below 0.05), and the residual width of scratch of cells in HIF-1α inhibitor group was wider at PSH 12 (P<0.05), and those of cells in HIF-1α stabilizer group were smaller at PSH 12 and 24 (with P values below 0.05). There was no obvious difference in the width of scratch in cells among the 7 groups (F=19.02, P>0.05). The protein levels of HIF-1α in ESCs of wild-type mice at PHH 0, 12, 24, and 48 were respectively 1.02±0.05, 2.56±0.09, 1.60±0.17, and 1.17±0.03. Compared with that at PHH 0, the protein level of HIF-1α at PHH 12 was significantly enhanced (P<0.01). It began to decline at PHH 24 but was still higher than that at PHH 0 (P<0.05), and the protein level of HIF-1α at PHH 48 was close to the normoxia level (P>0.05). Compared with those of cells in pure hypoxia group, the mRNA expressions of P311 of cells in HIF-1α inhibitor group were significantly decreased at each time point (with P values below 0.05), and those in HIF-1α stabilizer group were significantly increased at PHH 12 and 24 (with P values below 0.05). Compared with those of cells in HIF-1α inhibitor group, the mRNA expressions of P311 of cells in DMSO control group and HIF-1α stabilizer group were significantly increased at PHH 0, 12, and 24 (with P values below 0.05). Compared with those of cells in pure hypoxia group, the expressions of P311 of cells in HIF-1α inhibitor group were significantly decreased at each time point (with P values below 0.05), while those in HIF-1α stabilizer group were significantly increased at PHH 12 and 24 (with P values below 0.05). Compared with those of cells in HIF-1α inhibitor group, the expressions of P311 of cells in DMSO control group and HIF-1α stabilizer group were significantly increased at PHH 12 and 24 (with P values below 0.05). (2) The results of HEK-293 cells. At PCH 0, there was no significant difference in the luciferase activity among cells of empty vector hypoxia group, P311 normoxia group, P311 hypoxia group, and P311 hypoxia+ HIF-1α inhibitor group (F=13.33, P>0.05). At PCH 12, the luciferase activity of cells in P311 hypoxia group was higher than that in empty vector hypoxia group (P<0.01). The luciferase activity of cells in hypoxia group was higher than that in P311 normoxia group (P<0.05), while that of cells in P311 hypoxia+ HIF-1α inhibitor group was lower than that in P311 hypoxia group (P<0.01).@*Conclusions@#HIF-1α may increase the migration of murine ESCs through inducing the expression of P311 at the early stage of hypoxia.
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<p><b>OBJECTIVE</b>To explore the interaction between P311 and transforming growth factor beta 1 (TGF-β1) in murine fibroblasts and its effect on the function of fibroblasts.</p><p><b>METHODS</b>Skin fibroblasts obtained from five neonatal P311 wild-type C57BL/6 mice and P311 gene knock-out C57BL/6 mice were cultured. The second passage of fibroblasts were used in the following experiments. All experiments were repeated for 3 times. (1) The fibroblasts of P311 wild-type mice were divided into blank control group and P311 over-expression group according to the random number table (the same grouping method below), with 36 wells in each group. Fibroblasts in blank control group were transfected with 10 μL control vector, and fibroblasts in P311 over-expression group were transfected with equal efficiency P311 expression adenovirus vector. After being cultured for 48 hours, the mRNA expression level of P311, and the mRNA and protein expression levels of TGF-β1, α-smooth muscle actin (α-SMA), and collagen type I of fibroblasts in both groups were determined with real-time fluorescent quantitative RT-PCR and Western blotting (the same detection methods below), respectively. (2) After cultured reaching the cell density of 80%-90%, the mRNA and protein expression levels of TGF-β1, α-SMA, and collagen type I of the fibroblasts of P311 wild-type mice and P311 gene knock-out mice, with 4 flasks in each type of fibroblasts, were determined. (3) The fibroblasts of P311 wild-type mice were divided into blank control group and 5, 10, 15, 20, and 25 ng/mL TGF-β1 groups after being starved treatment with DMEM medium containing 1% FBS for 3 hours, with 2 flasks in each group. Fibroblasts in blank control group were routinely cultured, while fibroblasts in the latter five groups were treated with 5, 10, 15, 20, and 25 ng/mL TGF-β1, respectively. After being cultured for 48 hours, the mRNA expression levels of P311 in fibroblasts of the six groups were determined. Another fibroblasts of P311 wild-type mice were divided into blank control group and 10 ng/mL TGF-β1 group, with 6 wells in each group, and the protein expression levels of P311 in both groups were determined by immunofluorescence staining. (4) The fibroblasts of P311 wild-type mice were divided into blank control group and 10 ng/mL TGF-β1 group after being starved treatment as above, with 2 flasks in each group, and fibroblasts in blank control group were routinely cultured, while fibroblasts in the latter group were treated with 10 ng/mL TGF-β1. After being cultured for 48 hours, the mRNA and protein expression levels of α-SMA and collagen type Ⅰ were determined. The fibroblasts of P311 gene knock-out mice were grouped and treated as above, and the mRNA and protein expression levels of α-SMA and collagen type I were determined. Data were processed with one-way analysis of variance and t test.</p><p><b>RESULTS</b>(1) The mRNA expression level of P311 of fibroblasts in P311 over-expression group was increased nearly 300 000-fold compared with that in blank control group (t=9.942, P<0.001). The mRNA expression levels of TGF-β1, α-SMA, and collagen type I of fibroblasts in P311 over-expression group, and the protein expression levels of pro-TGF-β1, activated TGF-β1, α-SMA, and collagen type I of fibroblasts in P311 over-expression group were significantly higher than those in blank control group (with t values from 8.192 to 49.090, P values below 0.01). (2) The mRNA expression levels of TGF-β1, α-SMA, and collagen type I in fibroblasts of P311gene knock-out mice were significantly lower than those in fibroblasts of P311 wild-type mice (with t values from 8.157 to 22.270, P values below 0.01). The protein expression levels of pro-TGF-β1, α-SMA, and collagen type I in fibroblasts of P311 gene knock-out mice were significantly lower than those in fibroblasts of P311 wild-type mice (with t values from 2.995 to 12.600, P<0.05 or P<0.01), and the protein expression levels of active TGF-β1 were similar in two types of fibroblasts (t=1.070, P>0.05). (3) The mRNA expression levels of P311 of fibroblasts in blank control group and 5, 10, 15, 20 and 25 ng/mL TGF-β1 groups were 1.28 ± 0.44, 3.61 ± 0.91, 6.64 ± 0.92, 6.58 ± 1.04, 1.79 ± 0.31, 0.16 ± 0.06, respectively. Compared to the mRNA expression level of P311 of fibroblasts in the blank control group, the mRNA expression levels of P311 of fibroblasts in 5 and 20 ng/mL TGF-β1 groups were similar (with t values respectively 2.302 and 0.955, P values above 0.05), while they were significantly higher in 10 and 15 ng/mL TGF-β1 groups (with t values respectively 5.630 and 4.710, P values below 0.001), and they were significantly lower in 25 ng/mL TGF-β1 group (t=2.509, P<0.01). The protein expression level of P311 of fibroblasts in 10 ng/mL group was higher than that in blank control group. (4) The mRNA and protein expression levels of α-SMA and collagen type I of fibroblasts of P311 wild-type mice in 10 ng/mL TGF-β1 group were significantly higher than those in blank control group (with t values from 3.523 to 14.290, P<0.05 or P<0.01). The mRNA and protein expression levels of α-SMA and collagen type I of fibroblasts of P311 gene knock-out mice in 10 ng/mL TGF-β1 group were significantly higher than those in blank control group (with t values from 4.895 to 14.870, P<0.05 or P<0.01).</p><p><b>CONCLUSIONS</b>The interaction between P311 and TGF-β1 in murine fibroblasts exists and it may enhance the differentiation of fibroblasts in combination.</p>
Subject(s)
Animals , Mice , Actins , Metabolism , Cell Differentiation , Cells, Cultured , Collagen Type I , Metabolism , Fibroblasts , Cell Biology , Metabolism , Mice, Inbred C57BL , Mice, Knockout , Nerve Tissue Proteins , Genetics , Metabolism , RNA, Messenger , Transfection , Transforming Growth Factor beta1 , MetabolismABSTRACT
Objective To evaluate the comparability of the detection results of different blood cells analysis systems in same hos-pital.Methods Referring to the Guideline for Comparability Verification of Quantitative Test Results in Medical Institutions,the comparability validation protocol was established.The EDTA-K2 anticoagulation fresh whole blood samples with proper concentra-tion were detected for 5 parameters of HGB,RBC,HCT,PLT and WBC by 4 systems (Sysmex XT-1800i,Sysmex XT-2000,Sys-mex XT-4000i and Mindray BC-5800).The range was calculated and the detection results consistency analysis was performed.Re-sults The acceptable standard of critical differentials was intended to be HGB3.5%,RBC3%,HCT3%,PLT12.5% and WBC 7.5%.The replication detection was at least 2 times and up to 5 times.The ranges of 3 concentrations after replication detection and sample comparison were 2.87%-6.29%,1.57%-2.99%,1.95%-4.77%,12.81%-25.74% and 6.72%-11.13% respective-ly.The ranges of RBC detection results in 4 systems were smaller than the critical differentials,the validation was passed.The ran-ges of HGB,HCT,PLT and WBC detection results in 4 systems all had the condition of more than the critical differentials,the vali-dation did not passed.After removing the test system with obvious bias,the validation of the detection results by other test systems was passed.Conclusion The RBC detection results by 4 systems have the comparability;the HGB,HCT and PLT detection results by Sysmex XT-1800i,Sysmex XT-2000i and Sysmex XT-4000i have the comparability;the WBC detection results by Sysmex XT-1800i,Sysmex XT-2000i and Mindray BC-5800 have the comparability.
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We took measures to construct sound learning mechanism and a set of effective learning system,which were connected tightly with various learning ways according to the idea of ‘ school-based ’ training.Goals of learning mechanism and the set of effective learning system were to improve teacher's learning ability,enhance knowledge and competence of medical college teachers and to construct learning teaching faculty.
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BACKGROUND: Previous studies have shown that bone marrow mesenchymal stem cells (BMSCs) may be a promising candidate for cell-replacement therapies for neurodegenerative diseases including Alzheimer's disease (AD). But, it is still unclear whether BMSCs have therapeutic effects on AD.OBJECTIVE: To investigate the influence of BMSC transplantation on ethology of rats with AD, and to find a potential strategy for the development of effective therapies for the treatment of AD.DESIGN, TIME AND SETTING: The randomized controlled study was performed at the Laboratory of the Department of Physiology and Pathophysiology, Medical College of Chinese People's Armed Police Force in March 2009.MATERIALS: A total of 40 healthy male Wistar rats aged 24 months were used in this study, weighing approximately 450 g.METHODS: The natural senile AD rat model was chosen by maze test. The AD rats were randomly divided into four groups with ten animals in each group. In the control group, rats were injected bilaterally with physiological saline into the hippocampus. In the BMSC transplantation group, rats received BMSCs. In the normoxic differentiation and transplantation group, rats received injection of BMSCs induced from neuron-like cells under normoxic condition. In the hypoxic differentiation and transplantation group, rats received injection of BMSCs induced from neuron-like cells under hypoxic condition.MAIN OUTCOME MEASURES: The learning and memory ability of the AD rats were detected by Y type maze test 8 weeks later.Memory ability was tested 48 hours after learning test.RESULTS: The learning and memory scores decreased in control group and increased in BMSC transplantation group, there were all not statistics significance compared with that before transplant treatment (P > 0.05). The learning and memory scores were all higher than before in normoxic differentiation and transplantation group and hypoxic differentiation and transplantation group. There was significant difference between control group and the other groups (P < 0.01).CONCLUSION: BMSC transplantation may improve the learning and memory ability of AD rats. Our results demonstrate that BMSCs not only play an important role in improving cognitive disturbance of AD rats. Oriented differentiated BMSC transplantation was superior to non-differentiated BMSCs. Transplantation of hypoxic differentiated BMSCs has obtained optimal outcomes.
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Objectlve To demonstrate the feasibility of three-dimensional(3D)spin-lattice relaxation time in the rotating frame(T1ρ)-weighted imaging of porcine patellar cartilage in vitro at 7.0 T and the measurement of T1ρ of agarose phantom and patellar cartilage.Methods All the MR Imaging experiments were performed on a 7.0 T Varian scanner using a 6.0-cm-diameter quadrature birdcage RF coil tuned to 300 MHz.A 3D spin-echo sequence with a self.compensating spin-lock pulse cluster was used to acquire 3D-T1ρ-weighted images.The time of spin-locking(TSL)was from 0 to 50 ms with an interval of 10 ms.Spin-lock power wag 440 Hz.3D-T1ρweighted imaging was done three times for 6 phantoms (concentration 1%t0 6%),as well as once for 8 porcine patellar cartilages in order to assess the reproducibility of this technique.Signal-to-noise ratio(SNR)was measured on the acquired images of both phantoms and patellar cartilages,which were tested for significance using Two-way ANOVA.The images were processed on Vnmr J workstation using home-built processing software to construct 3 D T1ρ maps.T1ρ values were calculated within manually drawn regions-of-interest(ROI),and differences between groups were tested for significance using analysis of variance(One-way ANOVA).Results T1ρ -weighted images with a shorter TSL had a higher SNR value,which measured between 48±8 and 95±8 in the global cartilage.Cartilage images had a higher SNR(TSL＜30 ms)compared to agarose phantoms and a lower SNR(TSL ＞30 ms)only compared to l%agarose phantorm T1ρ relaxation times in agarose phantoms increased as agarose concentrations decreased in global regions.The CV of T1ρ in agarose phantoms was less than 10%.Global and regional analyses of patellar cartilage T1ρ were 68.9±6.3 ms,80.7±12.8 ms,65.7±7.0 ms,82.4±7.7 ms,and 69.7±6.4 ms in the global cartilage,the superficial layer,the transitional layer,the deep layer,and the calcified layer,respectively.T1ρ in the superficial and deep layer was significantly higher than in the transitional,calcified layer and global cartilage(F=6.436,P＜0.05).Conclusions The present study demonstrates the feasibihty of 3D-T1ρ-weighted imaging of porcine patellar cartilage at 7.0 T with hish image quality.T1ρ maps can be used to quantify the laminar structures in 3D-T1ρ-weighted images of articular cartilage,which pave the way to evaluate early osteoarthritis and cartilage regeneration.
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ObjectiveTo compare the sweep pattern visual evoked response acuity SPVER-A with the logarithmic visual acuity.MethodsSPVER-A and logarithmic visual acuity were measured and compared in 50 people. 8-squared vertical gratings were presented as stimuli continuously.The responses were averaged and displayed through the discrete fourier transform on the monitor display.The SPVER acuity was determined by extrapolating the SPVER amplitude-spatial frequency function to baseline.ResultsThe overall correlation was r=0.699 between the SPVER-A and logarithmic visual acuity.Underestimation of acuity by the SPVER when the logarithmic visual acuity is better than 0.5.On the other hand,when the logarithmic visual acuity is worse than 0.5 the SPVER-A was overestimated.ConclusionSPVER-A has a good correlation to logarithmic acuity.The difference between two methods may indicate that the grating acuity and logarithmic /visual acuity activate different system.
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Objective:To evaluate p53,c erbB 2,p21 and nm23 oncoprotein expression in diagnosis and treatment of gastric cancer.Methods:The expression of p53,c erbB 2,p21 and nm23 oncoproteins was detected with immunohistochemical method in 63 surgically resected specimens and its endoscopic biopsy specimens of gastric cancer.Results:The positive rates of p53,c erbB 2,p21 and nm23 oncoprotein expression were 37 6%～46 2%,34 6%～56 8%,37 8%～61 5%,70 3%～30 8% respectively.Oncoprotein expression was not observed in non tumor endoscopic biopsy specimens.The expression of c erbB 2,p21 was correlated with grade of tumor differentiation and the expression of p21,nm23 oncoproteins was correlated with the depth of tumor invasion and clinical different stage,namely tumor metastasis.Positive rates of p53,c erbB 2,p21 and nm23 oncoproteins between biopsy and resected specimens was of coincidence.Conclusion:Determination of p53,c erbB 2,p21 and nm23 oncoprotein expression was might be useful in diagnosis and treatment of gastric cancer,differentiating benign and malignant tumor and clinical stages,of gastric cancer. [
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HPLC maps of Panax quinqueflius L. and sundried Ginseng (P. ginseng) were compared in detail. Theirdifferences together with the amount of Rb1 ginsenoside present in the samples can be used to differentiate thetwo kinds of Ginseng with ease and accuracy.