This study aimed to study protein expression in diabetic nephropathy (DN) rats and stem cell tissue repair based on nanomembrane concentration technology. Based on the polymer porous self-assembled nanomembrane technology, the content of total protein and albumin in the serum of rats in the control and experimental groups were measured. The obtained images were adopted to analyze the expression of cytoplasmic proteins and membrane proteins, and then the mechanism of stem cell tissue repair function was studied. The results showed that at 56 weeks of age, in contrast to control group, the total protein content in the serum of the experimental group evidently decreased. At 36 weeks of age, the cytoplasmic protein samples of diabetic rats in the experimental group were subjected to three two-dimensional protein electrophoresis. It was found that there were about 701 spots in each gel, and the matching rate was about 87.5%. In contrast to control group, 16 cytoplasmic proteins and 23 membrane proteins of the experimental group changed. The results of the differentially expressed protein analysis indicated that the change trends of protein spots. Based on polymer porous self-assembled nano-film technology, the expression of differential proteins in DN rats was analyzed, which would supplement new research assistance to the mechanism of DN.

Objective To compare the effectiveness of traditional and hybrid teaching strategies in pathophysiology and to conduct a survey of students' opinions about the hybrid teaching strategy. Methods A hybrid pathophysiology course was developed by combining traditional lectures, case- or problem-based learning, group discussion and several quizzes. A total of 167 students were assigned to the hybrid teaching group and 118 students assigned to the traditional lecture group. Results Compared with students who received traditional lectures, no students in the hybrid teaching class failed the final examination. The percentage of students with high scores was significantly higher in the hybrid teaching class. In addition, 73.7% of students in the hybrid teaching class expressed substantial interest in pathophysiology during the course, and 83% of these students felt they had received essential training and acquired the ability to solve clinical case problems. Conclusion The hybrid teaching strategy is an advanced approach that encourages students to actively learn teaching materials and solve practical clinical problems, and that promotes student interest in pathophysiology.

目的 对实验操作进行量化考核,构建科学的病理生理学机能实验考核体系.方法 选择2015级麻醉学专业160名学生为考核对象,在小范围建立新的机能实验操作技能量化考核体系,结合平时考核,作为其最终的实验考核成绩,并对其可行性和优势进行了分析评估.结果 参与考核的同学实验操作合格率98.75％,优秀率82.5％.调查问卷回收率100％,其中,98.75％ 认为新的实验考核测评模式对本人的操作能力、解决问题能力有帮助和提高,94.38％ 认为新的实验考核测评模式能够促进团队合作,有利于协作精神的培养,91.88％认为新的实验考核测评模式效果良好.结论 新的实验考核测评模式对学生的操作能力、解决问题能力、团队合作、协作精神培养方面有显著作用,可有效促进学生的机能课程学习和教师的教学活动,从而促进学生的长远发展和病理生理学教学质量的提高.

Aims The main objective was to investigate the effects of the transient receptor potential cation channel subfamily V member 1 (TRPV1) on nerve regeneration following sciatic transection injury by functional blockage of TRPV1 using AMG-517, a specific blocker of TRPV1. Methods AMG-517 was injected into the area surrounding ipsilateral lumbar dorsal root ganglia 30 min after unilateral sciatic nerve transection. The number of sciatic axons and the expression of growth-associated protein-43 (GAP-43) and glial fibrillary acidic protein was examined using semithin sections, Western blot, and immunofluorescence analyses. Results Blockage of TRPV1 with AMG-517 markedly promoted axonal regeneration, especially at two weeks after sciatic injury; the number of axons was similar to the uninjured control group. After sciatic nerve transection, expression of glial fibrillary acidic protein was decreased and GAP-43 was increased at the proximal stump. However, the expression of both glial fibrillary acidic protein and GAP-43 increased significantly in AMG-517-treated groups. Conclusions TRPV1 may be an important therapeutic target to promote peripheral nerve regeneration after injury.

病理生理学作为连接基础与临床的主干桥梁学科,位于医学教育的中期阶段,在教学过程中,基于团队的讨论式教学模式(team-based learning,TBL),引入临床案例分析有利于对病理生理学知识点的理解以及基础医学和临床医学的融通.一份合格的教学案例应该符合地方院校的特色、病种单一,满足不同阶段医学生的需要,并且在教师的组织、引导下得以恰当的应用.

Objective To investigate the effect of recombinant botulinum neurotoxin serotype A heavy chain (BoNT/A heavy chain)on local proteins which are related to nerve growth after spinal cord injury in rats,and to get some experimental evidence to explain the mechanism of BoNT/A heavy chain in stimulating neuritogenesis. Methods Recombinant botulinum neurotoxin serotype A heavy chain was applied locally or intrathecally to rats with ipsilateral semi-dissociated lumbar spinal injury. Local spinal tissue was extracted for general protein expression by two dimension electrophoresis plus nitrate silver staining after different time period of injury. Based on the results of 2-D gel electrophoresis,growth-associated protein 43(GAP-43)and of superior cervical ganglion 10(SCG 10)were selected to examine the changes of their expression and distribution features under BoNT/A heavy chain administration using SDS-PAGE,western blot and immunofluorescence. Results (1)The model of spinal cord injury(SCI)in this study was an ipsilateral semi-dissociated lumbar SCI in rat. The rats showed obvious motor and sensory dysfunction in the ipsilateral hind limb.(2)The results from 2-D gel electrophoresis plus nitrate silver staining showed that the administration of BoNT/A heavy chain based on SCI altered the local protein expression pattern. The decrease or increase in the expression of some protein dots /dots group was clearly seen after single SCI. However, these changes were transformed by BoNT/A heavy chain treatment,which appeared as a reversed pattern turning toward that in control group or further increased expression upon SCI,such as the dots located respectively at 35-45 kDa and 18-25 kDa level,pI between 5-7. In addition,the expression of the two dots located at the level as above increased after SCI only, and showed further increase in their expression with BoNT/A heavy chain intervention.(3)The changes of selective GAP-43 and SCG 10 expression and distribution by western blot and immunofluorescence indicated that the administration of BONT/A heavy chain based on SCI amplified the expression of GAP-43 and SCG 10(P < 0.05). Meanwhile,the positive immuonfluorescent staining for both GAP-43 and SCG 10 mainly distributed nearby the proximal area of injury, both cytoplasm and neuronal processes were positively stained. Conclusions Intrathecal delivery of BoNT/A heavy chain increases the expression of growth-associated proteins GAP 43 and SCG 10 after SCI in rats.

Objective To investigate the effect and molecular mechanism of botulinum neurotoxin serotype A (BoNT/A) heavy chain on neuron regeneration. Methods Cell culture, rats, immunofluorescence, SDS-PAGE and western blot, etc. were adopted in this study to explore the alterations of histone-3 acetylation (acetyl-H3 ) by local treatment of BoNT/A heavy chain to spinal cord injury (SCI) in rats (in vivo) or by adding it into cell culture (in vitro). Meanwhile, the relevance of acetyl-H3 to neurite out-growth based on SCI and cell culture with BoNT/A heavy chain application was approached as well. Results The application of BoNT/A heavy chain to cultured Neuro-2a cells increased the level of H3 acetylation. The increase of H3 acetylation was paralleled with the growth of neuritogenesis. Also, the neuronal treatment of BoNT/A heavy chain to SCI promoted the re-growth of neuronal processes surrounding the lesions. The growth of neuronal processes was positively correlated to the level of H3 acetylation. During the periods of BoNT/A heavy chain treatment in vivo or in vitro, the increase of H3 acetylation showed two peaks. Conclusions BoNT/A heavy chain increased the H3 acetylation, which might be one of its neuritogenic mechanisms.

(1) Background: The botulinum toxin A (BoNT-A) heavy chain (HC) can stimulate the growth of primary motor neurites. (2) Methods: A recombinant BoNT/A HC was injected locally plus interval intrathecal catheter of BoNT/A HC to rats with ipsilateral semi-dissociated lumbar spinal cord injuries (SCIs). First, 2D gel with a silver nitrate stain was applied to detect the general pattern of protein expression. Growth associated protein 43 (GAP-43) and superior cervical ganglion 10 (SCG10) were chosen to represent the altered proteins, based on their molecular weight and pI, and were used to further detect their expression. Meanwhile, the neuronal processes were measured. The measurements of thermal hyperalgesia and grasp power at the ipsilateral hindlimb were used to evaluate spinal sensory and motor function, respectively. (3) Results: The local injection of BoNT/A HC followed by its intrathecal catheter intervally altered the spinal protein expression pattern after an SCI; protein expression was similar to normal levels or displayed a remarkable increase. The changes in the expression and distribution of phosphorylated growth associated protein 43(p-GAP 43) and superior cervical ganglion 10 (SCG 10) indicated that the administration of BoNT/A HC to the SCI significantly amplified the expression of p-GAP43 and SCG10 (p < 0.05). Meanwhile, the positive immunofluorescent staining for both p-GAP43 and SCG10 was mainly present near the rostral aspect of the injury, both in the cytoplasm and the neuronal processes. Moreover, the outgrowth of neurites was stimulated by the BoNT/A HC treatment; this was evident from the increase in neurite length, number of branches and the percentage of cells with neuronal processes. The results from the spinal function tests suggested that the BoNT/A HC did not affect sensation, but had a large role in improving the ipsilateral hindlimb grasp power (p < 0.05). (4) Conclusions: The local injection with the intermittent intrathecal administration of BoNT/A heavy chain to rats with SCI increased the local expression of GAP-43 and SCG 10, which might be affiliated with the regeneration of neuronal processes surrounding the injury, and might also be favorable to the relief of spinal motor dysfunction.

目的:为了完善病理生理学现有的教学评估体系、结合TBL教学模式,在实习课中进行学生互评体系的尝试.方法:评估流程包括:学生以团队为单位;对教师所提供病例进行思考;就病例内容进行团队间的互相发问及相互评分;教师根据各团队在问答中的表现进行综合评分.结果:团队通过角色互换,完成互评.

Myelin-associated glycoprotein (MAG) inhibits the growth of neurites from nerve cells. Extraction and purification of MAG require complex operations; therefore, we attempted to determine whether commercially available MAG-Fc can replace endogenous MAG for research purposes. Immunofluorescence using specific antibodies against MAG, Nogo receptor (NgR) and paired immunoglobulin-like receptor B (PirB) was used to determine whether MAG-Fc can be endocytosed by neuro-2a cells. In addition, neurite outgrowth of neuro-2a cells treated with different doses of MAG-Fc was evaluated. Enzyme linked immunosorbent assays were used to measure RhoA activity. Western blot assays were conducted to assess Rho-associated protein kinase (ROCK) phosphorylation. Neuro-2a cells expressed NgR and PirB, and MAG-Fc could be endocytosed by binding to NgR and PirB. This activated intracellular signaling pathways to increase RhoA activity and ROCK phosphorylation, ultimately inhibiting neurite outgrowth. These findings not only verify that MAG-Fc can inhibit the growth of neural neurites by activating RhoA signaling pathways, similarly to endogenous MAG, but also clearly demonstrate that commercial MAG-Fc is suitable for experimental studies of neurite outgrowth.

目的:探讨人工重组A型肉毒毒素重链(BoNT/A重链)对大鼠脊髓损伤局部颈上神经节蛋白10(SCG10)表达、神经突起再生以及损伤侧后肢感觉及运动功能的影响,为阐明BoNT/A重链促神经突起生长的作用提供实验依据.方法:大鼠单侧腰段脊髓横断损伤模型基础上局部及鞘内给予BoNT/A重链;于用药后不同时间提取局部蛋白进行双向电泳,并根据双向电泳提示的蛋白点群变化的分子量和等电点,选取SCG10为目标蛋白行Western Blot检测及神经突起测定;通过抓力和热痛敏实验测试损伤侧后肢的运动和感觉功能情况.结果:①BoNT/A重链可干预脊髓损伤后局部蛋白谱的变化,MW位于18-25kDa、等电点在5-7范围的蛋白点可以作为蛋白点群变化的代表之一;②作为分子量位于18-25kDa/等电点在5-7的蛋白成分之一的SCG10在单纯损伤时较正常对照组其表达有所增高,给予BoNT/A重链后,SCG10的表达显著增强(P＜0.05);③免疫荧光结果显示:单纯损伤时SCG10的阳性表达主要分布于损伤周边细胞的胞体,应用BoNT/A重链后,SCG10阳性分布于胞浆和细胞突起且以损伤近头端的周边区域较为显著;④突起测定结果显示:BoNT/A重链可促进脊髓损伤周边神经细胞突起增长,包括突起数目增多及含有突起的神经细胞百分比增多(P＜0.05);⑤给予BoNT/A重链后,损伤侧后肢肌力明显优于单纯损伤组(P＜0.05),但感觉功能未见明显改善.结论:脊髓损伤局部给予BoNT/A重链可促进生长相关蛋白SCG10的表达并促进损伤周边神经细胞突起生长,改善肌肉抓力.BoNT/A重链有助于脊髓损伤后的再生修复.

目的:了解中小门诊医疗垃圾的处理现状,并对定点试行的医疗垃圾车辆集中收集的处理方法与传统的挂靠政策进行比较.方法:选择分布于太原市两个城区、执行不同医疗垃圾采集政策的30家医疗诊所(中小型门诊)进行实地走访调查,并对所调查结果进行统计学分析.结果:采用垃圾车辆集中收集的A城区各诊所中,能将医疗垃圾与生活垃圾严格分类的医疗机构占93％;而采纳政府挂靠政策的B城区内的医疗诊所,医疗垃圾和生活垃圾达到有效分类者占83％;两城区医疗垃圾均能做到具体分类.对于装医疗垃圾的容器或包装物,A城区的合格率为87％,B城区为63％.医疗垃圾储存过期率A城区为3％,B城区为93％.结论:太原市部分城区中小门诊医疗垃圾处理方法存在较大问题.综合来看,医疗垃圾车辆分片集中收集优于挂靠政策,有利于医疗垃圾的规范化处理.

文章主要对目前国内医学院校双语教学及专业英语教学设置、教学人员的专业英语水平及实施现状进行了分析;结合专业英语及双语教学的特点提出了专业英语课与专业课双语教学两者在教与学之间的关联性:前者通过专业知识学习英语,后者用英语阐述医学知识,二者密切相关;文章还就如何在今后的医学教学实践中将二者融为一体提出了方法和建议.

文章就目前研究生专业英语的现状进行了剖析,对存在的问题进行了归纳总结.同时,对医学英语术语学的特征及内涵进行概要介绍,结合现状强调了医学英语术语学在我国研究生教学中的作用及其意义并对今后教研室学习过程中如何进行专业英语及医学英语术语学的融合及师资培训提出了建议.

Objective: To observe the effect of the expressive or functional blockage of TRPV1 on nerve regeneration after sciatic trans-section injury. Methods: AMG-517, a kind of TRPV1 inhibitor, was injected into the surrounding area of the ipsilateral lumbar dorsal root ganglia while unilateral sciatic nerve was transected. A total of 24 healthy male Sprague-Dawley rats were divided into 4 groups: control group, injury only group, injury+ AMG-517 150 μg/kg group, injury+ AMG-517 300 μg/kg group. The injury only group was injected the same volume of medium. The release of CGRP from dorsal-horn of spinal cord, the number of axons at proximal stem of sciatic nerve after transection, and the expression of TRPV1 in dorsal root ganglion were detected using the methods of ELISA, Western blot and semi-thin section (1 μm)- toluidine blue staining 2 weeks after injury. Results: The release of CGRP in lumbar spinal dorsal horn was obviously decreased after AMG-517 treatment, which was the evidence of TRPV1 functional inhibition. CGRP in the control group was 0.15 ng/g, the injury only group 0.17 ng/g, AMG-517 150 μg/kg group 0.09 ng/g, and AMG-517 300 μg/kg group 0.11 ng/g(P<0.01). The number of axons which were myelinated or unmyelinated increased after the TRPV1 was inhibited by AMG-517(P<0.01). In addition, the injection of AMG-517 into surrounding dorsal root ganglion decreased the expression of TRPV1 in dorsal root ganglion(P<0.01). Conclusions: Over expression or activation of TRPV1 after periphery nerve injury has negative effect on nerve regeneration in fact; Inhibiting the over-expression or over-activation of TRPV1 after nerve injury facilitates axonal regeneration and nerve repair.

病理生理学是一门研究疾病发生、发展、转归规律和机制的学科,能够帮助医学生建立初步的临床思维.对病理生理学传统教学方式和考核方式进行尝试性调整,在本硕班采用改良的TBL教学模式以及过程性评价和终结性评价相结合的考核模式,提高了学生的自主学习能力和解决问题的综合能力,从而提高了教学质量.

BACKGROUND:Botulinum neurotoxins are produced by Clostridium botulinum bacteria. There are eight serologically distinct botulinum neurotoxin isoforms (serotypes A-H). Currently, botulinum neurotoxin serotype A (BoNT⁄A) is commonly used for the treatment of many disorders, such as hyperactive musculoskeletal disorders, dystonia, and pain. However, the effectiveness of BoNT⁄A for pain alleviation and the mechanisms that mediate the analgesic effects of BoNT⁄A remain unclear. To define the antinociceptive mechanisms by which BoNT/A functions, the interactions between BoNT⁄A and the transient receptor potential vanilloid subfamily 1 (TRPV1) were investigated using immunofluorescence, co-immunoprecipitation, and western blot analysis in primary mouse embryonic dorsal root ganglion neuronal cultures.RESULTS:1) Three-week-old cultured dorsal root ganglion neurons highly expressed transient TRPV1, synaptic vesicle 2A (SV2A) and synaptosomal-associated protein 25 (SNAP-25). SV2A and SNAP-25 are the binding receptor and target protein, respectively, of BoNT⁄A. 2) TRPV1 colocalized with both BoNT⁄A and cleaved SNAP-25 when BoNT⁄A was added to dorsal root ganglia neuronal cultures. 3) After 24 hours of BoNT⁄A treatment (1 nmol⁄l), both TRPV1 and BoNT⁄A positive bands were detected in western blots of immunoprecipitated pellets. 4) Blocking TRPV1 with a specific antibody decreased the cleavage of SNAP-25 by BoNT⁄A.CONCLUSION:BoNT/A interacts with TRPV1 both structurally and functionally in cultured mouse embryonic dorsal root ganglion neurons. These results suggest that an alternative mechanism is used by BoNT⁄A to mediate pain relief.

AIM:To observe the effect of botulinum neurotoxin type A heavy chain ( BoNT/A HC) on the pat-tern of spinal protein expression by intrathecal injection after spinal cord injury in rats , and to explore the role of BoNT/A HC intervention in spinal protein expression and some of its mechanisms in nerve regeneration after injury .METHODS:The model of unilateral lumbar spinal cord injury was established .The effects of BoNT/A HC intervention at different doses (2 μg, 4 μg, 6 μg and 8 μg) on the general pattern of protein expression in the spinal cord tissues at the injury site and the cranial part adjacent to the injury site was measured and evaluated by SDS-PAGE and Coomassie brilliant blue staining first, and then by two-dimensional SDS-PAGE.RESULTS:The histological structure of the ipsilateral side of lumbar spi-nal cord showed obvious destruction and degradation , mainly affecting both gray and white matter of the left side of the cord.The result of SDS-PAGE with Coomassie brilliant blue staining from injured spinal cord tissue displayed that the ex-pression of some proteins after one-time BoNT/A HC treatment appeared obviously different from that without BoNT /A HC treatment.Moreover, the pattern of the protein expression affected by BoNT/A HC was similar to that of the normal spinal cord.The more detail information from two-dimensional SDS-PAGE indicated that more than 10 proteins with different mo-lecular weight and isoelectronic points were differentially expressed at day 2 and day 20 after local injection of 6μg BoNT/A HC.This altered expression actually appeared a tendency toward the pattern shown in normal group .CONCLUSION:The immediate application of BoNT/A HC at the injury site after unilateral lumbar spinal cord injury is able to affect the pattern of local protein expression .The altered protein expression by injury could be reversed back to normal or approxi -mately normal by local BoNT/A HC administration.