全面开展线上教学的尝试及对中医药类金课建设的思考

线上教学一直是高等教育教学改革的重点,在慕课、微课、精品在线开放课程等线上教学资源库建设的基础上,线上教学手段和方法更趋于完善,但全面使用线上手段完成教学仍处在尝试阶段.面对特殊的情况,充分利用前期网络资源库,做好教师与学生的角色定位,切实落实完善教学质量的管理和监督,充分开展思政教育,是我校线上教育教学模式变革的新方向.同时,面对教育部提出的"打造金课,淘汰水课"的课程建设口号,我校以线上教学为切入点深挖学科特点,紧密结合专业特色的尝试,更是为探索中医药类金课建设打下了基础.     

Effects of acupuncture plus mild hypothermia on apoptosis-related factors in rats with cerebral ischemia-reperfusion

Objective

To investigate the effect of acupuncture plus mild hypothermia on neurological function impairment score, cerebral infarct size and apoptosis-related factors in cerebral ischemia reperfusion injury (CIRI) rats.

Methods

Sixty healthy male Sprague-Dawley (SD) rats were routinely reared for 1 week. Ten rats were randomly selected as the sham operation group and 10 rats as the blank control group, while the remaining 40 rats were subjected to preparing the middle cerebral artery occlusion (MCAO) model by modified filament occlusion method. The 40 MCAO rats were further randomly divided into a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, with 10 rats in each group. Rats in the sham operation group, the blank control group and the model group did not accept treatment except binding; rats in the acupuncture group received acupuncture treatment; rats in the mild hypothermia group received mild hypothermia treatment; rats in the acupuncture plus mild hypothermia group received acupuncture and mild hypothermia treatment. 72 h after the treatment, neurological function impairment score was performed; the infarct area ratio was determined by 2,3,5-tripheyl tetrazolium chloride (TTC) staining; apoptosis of brain cells was observed by TUNEL method; the expressions of Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry.

Results

Compared with the blank control group and the sham operation group, the neurological function impairment score, cerebral infarct area ratio, apoptosis, and the expressions of Bax and Caspase-3 in the model group were significantly increased, while the expression of Bcl-2 was significantly decreased, and there were significant between-group differences (all P<0.05). After the treatment, there were statistically significant differences among the treatment groups in the neurological function impairment score, cerebral infarct area ratio and apoptosis in the ischemic side of rats, as well as the expressions of Bcl-2, Bax and Caspase-3 (all P<0.05), and from the figures, tables and statistical analysis, it was found that a better tendency in the acupuncture plus mild hypothermia group than the acupuncture group or mild hypothermia group.

Conclusion

Acupuncture plus mild hypothermia can protect the brain cells by improving neurological function impairment, decreasing cerebral infarct area ratio, reducing the number of apoptotic cells in the ischemic area and regulating the expressions of apoptosis related proteins to inhibit apoptosis.

     

针刺大椎、百会、人中穴对脑缺血再灌注损伤大鼠脑组织p-VEGF蛋白表达的影响

目的观察针刺大椎、百会、人中穴对脑缺血再灌注损伤大鼠脑组织p-VEGF蛋白表达的影响,探讨针刺对脑缺血再灌注损伤的部分作用机制。方法从60只大鼠中随机选择24只,再将之随机分成正常组和假手术组,12只/组,其余36只大鼠参照Zea Longa线栓法制备大脑中动脉栓塞模型,造模成功后再随机分为模型组、针刺组、依达拉奉组,12只/组。造模大鼠生命体征平稳后,正常组、假手术组及模型组大鼠只捆绑不针刺;针刺组大鼠针刺大椎、百会、人中三穴,留针30 min;依达拉奉组大鼠按3 mg/kg(稀释至1mL)腹腔注射依达拉奉,均为每12小时1次;首次治疗前及末次治疗后,对大鼠进行神经功能缺损评分;6次治疗完成后处死大鼠,行TTC染色观察脑梗死面积比,采用Western blot法检测p-VEGF蛋白表达水平。结果与正常组及假手术组比较,模型组大鼠神经功能缺损评分及脑梗死面积比显著上升(P0.01),提示模型制备成功。与模型组比较,针刺组及依达拉奉组大鼠神经功能缺损评分及脑梗死面积比下降(P0.01);同时,与模型组比较,针刺组及依达拉奉组大鼠p-VEGF蛋白表达上升(P0.01)。针刺组与依达拉奉组比较,大鼠神经功能缺损评分、脑梗死面积比及p-VEGF蛋白表达均无差异(P0.05)。结论针刺大椎、百会、人中穴能明显减轻脑缺血再灌注损伤,且其机制可能与上调VEGF蛋白磷酸化表达,进而促进血管新生有关。     

针刺大椎、百会、人中穴对大鼠脑缺血再灌注损伤后不同时间点海马脑红蛋白表达的影响

目的观察针刺大椎、百会、人中穴对大鼠脑缺血再灌注损伤后不同时间点海马脑红蛋白(neuroglobin,Ngb)表达的影响,探讨其脑保护作用的部分作用机制。方法采用线栓法制备MCAO模型,将40只雄性SD大鼠随机分为假手术组、模型组、对照点组、穴位组4大组,每组再根据再灌注后时间分为24 h、72 h组,每组5只。完成治疗后,先行神经功能缺损评分再处死大鼠,再采用Western Blot法检测大鼠缺血侧海马Ngb的表达水平。结果神经功能缺损评分:与假手术组比较,模型组、对照点组及穴位组神经功能缺损评分均升高,差异有统计学意义(P0.05,P0.01);模型组、对照点组、穴位组三组神经功能缺损评分差异无统计学意义(P0.05)。与24 h组比较,72 h模型组神经功能缺损评分差异无统计学意义(P0.05);72 h对照点组及穴位组神经功能缺损评分均下降,差异有统计学意义(P0.05)。Ngb:与假手术组比较,模型组Ngb的表达水平明显降低(P0.01);与模型组比较,对照点组及穴位组Ngb的表达水平均明显升高(P0.01);与对照点组比较,穴位组Ngb的表达水平有升高,但差异无统计学意义(P0.05)。与24 h组比较,72 h假手术组、模型组及对照点组Ngb的表达水平均有下降,但差异无统计学意义(P0.05);72 h穴位组Ngb的表达水平下降明显,差异有统计学意义(P0.05)。结论针刺能降低脑缺血再灌注损伤大鼠神经功能缺损评分,并可上调海马Ngb的表达水平,从而实现脑保护作用。     

中医临床专业学位研究生培养实践与探讨

基于中医临床专业学位研究生的培养现状探讨行之有效的培养对策。（1）重视经典学习,加强中医文化素养;（3）加强传统文化教育,培养良好医德医风;（3）加强临床能力考核,充分发挥考核的杠杆作用。     

应用蛋白芯片技术筛选针刺对脑缺血再灌注损伤大鼠的凋亡相关蛋白

目的应用蛋白芯片技术筛选针刺对缺血再灌注损伤（CIRI）大鼠脑组织凋亡相关蛋白。方法 SD健康大鼠40只随机分为4组：假手术组、模型组、针刺对照点组（均取穴左侧旁开0.3 cm的非经非穴点）、针刺穴位组（针刺大椎、百会、人中）,每组10只,各组大鼠6次处理后取左侧海马脑组织,采用720磷酸化抗体蛋白芯片检测处理后的脑组织细胞信号蛋白磷酸化的变化,然后筛选各组上调、下调显著（磷酸化水平变化上调≥1.5倍,下调≤0.67倍）且与凋亡相关的蛋白。结果与假手术组比较,模型组上调≥1.5倍与凋亡相关的蛋白8种,下调≤0.67倍的蛋白4种;与模型组比较,对照点组上调≥1.5倍与凋亡相关的蛋白5个,下调≤0.67倍的蛋白8个;穴位组上调≥1.5倍与凋亡相关的蛋白2个,下调≤0.67倍的蛋白11个,其中下调模型组中上调的蛋白3种。结论针刺大椎、百会、人中（穴）对CIRI大鼠通过调整多个凋亡相关的蛋白表达变化抑制凋亡实现脑保护作用。     

温针百会及颈夹脊穴治疗牙龈出血验案1则

正牙龈出血是指牙龈自发性或因轻微刺激而导致的出血现象,临床上常见的诱发因素有牙菌斑、牙石等局部刺激引发的牙周组织炎症,凝血机制障碍、血流动力学改变以及心脑血管疾病。该病属中医学"牙衄"范畴,根据其病因病机主要分为气血亏虚、脾胃湿热、脾气虚弱、肾阴亏损等型。治疗上,西医主要采取局部压迫止血、去除局部刺激因素、     

电针不同穴组对溃疡性结肠炎模型大鼠血清IL-6、IL-10含量的影响

目的:观察"俞募配穴"、"合募配穴"两种针刺配穴法治疗大肠腑病的效应差异,探讨"合治内府"之"合"的含义。方法:40只Wistar大鼠随机分为5组:空白组、模型组、曲池天枢组、天枢上巨虚组、天枢大肠俞组。酶联免疫吸附法(ELISA法)测定各组大鼠血清白介素6(IL-6)、白介素10(IL-10)含量的变化。结果:与空白组比较,模型组大鼠血清IL-6明显升高,IL-10明显降低,差异有统计学意义(P<0.01);模型组比较,各治疗组IL-6明显降低、IL-10明显升高,差异有统计学意义(P<0.01);与曲池天枢组比较,其它治疗组IL-6明显降低,IL-10明显升高,差异有统计学意义(P<0.05或<0.01);天枢大肠俞组与天枢上巨虚组比较,IL-6含量明显降低,差异有统计学意义(P<0.05)。结论:各治疗组对UC大鼠均有一定治疗作用,其中天枢配上巨虚组效果最佳,即下合穴配募穴治疗大肠腑病的效应优于俞募配穴及上合穴配募穴;"合治内府"之合穴应指"下合穴"。     

Effect of acupuncture plus mild hypothermia on MAPK/ERK pathway of brain tissues in rats with cerebral ischemia-reperfusion injury

Objective

To observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury (CIRI), and the influence on protein expression levels of phosphorylated Raf-1, MEK-2 and ERK1/2 in the mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinases (ERK) pathway, and to explore the mechanism of acupuncture plus mild hypothermia therapy for the ischemic stroke.

Methods

Ninety Sprague-Dawley (SD) rats were randomly divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, 15 rats in each group. Except the rats in the blank control group, the remaining rats were used to prepare the middle cerebral artery occlusion (MCAO) models according to the modified occlusion method using lines, while only the occlusion lines were inserted without blocking the brain arteries of rats in the sham operation group. When the vital signs of rats were stable, rats in the blank control group did not receive any intervention; rats in the sham operation group and the model group received fastening without treatment; rats in the acupuncture group, the mild hypothermia group, and the acupuncture plus mild hypothermia group were treated with the corresponding therapeutic methods. 72 h later, observed neurologic injury score, evaluated infarction area ratio by 2,3,5-tripheyl tetrazolium chloride (TTC) staining, determined apoptosis by TUNEL assay, and measured the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels in rat ischemic hippocampal tissues by Western blot assay.

Results

Compared with the blank control group and the sham operation group, after modeling, the neurologic injury score, infarction area ratio and apoptotic cells were increased, and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly increased in the model group; the differences were statistically significant (P<0.05 or P<0.01). Compared with the model group, after acupuncture or mild hypothermia therapy, neurologic injury score and infarction area ratio were decreased; apoptotic cells and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly decreased; the differences were statistically significant (P<0.05 or P<0.01). Compared with the acupuncture group, neurologic injury score and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were decreased in the acupuncture plus mild hypothermia group; differences between the groups were statistically significant (P<0.05 or P<0.01). Compared with the mild hypothermia group, phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels decreased in the acupuncture plus mild hypothermia group, and differences were statistically significant (P<0.01).

Conclusion

Acupuncture or mild hypothermia therapy can improve neurologic injury, reduce infarction area and apoptosis, which brought about protective effect on the brain tissues, in the MCAO model. The protective effect of acupuncture plus mild hypothermia group is the strongest. The mechanism may involve the MAPK/ERK pathway, by reducing the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels.

     

Effects of acupuncture on the number of associated protein phosphorylation in brain tissues of MCAO rats based on protein microarray technique

Objective

To investigate the effects of acupuncture on the number of associated phosphorylated proteins in brain tissues of middle cerebral artery occlusion (MCAO) rats, based on the protein microarray technique.

Methods

The MCAO model was prepared according to the modified occlusion method using occlusion lines. Forty healthy Sprague-Dawley (SD) rats were randomly divided into 4 groups using the lottery method: a sham operation group, a model group, a control point group and an acupoint group, with 10 rats in each group. Rats in the sham operation group and the model group only received binding without acupuncture. Rats in the acupoint group received acupuncture at Dazhui (GV 14), Baihui (GV 20) and Shuigou (GV 25); rats in the control point group received acupuncture at non-acupoint control points. The needle was twisted once for 1 min after insertion and another time in the middle of the 30 min needle retaining. Acupuncture was conducted once every 12 h for 6 consecutive times. At the end of the experiment, the neurological impairment score was collected, and cells of the ischemic brain tissues were extracted. The protein phosphorylation of the related signaling was detected using the 720 phosphorylated antibody microarray technique, and the differentially expressed proteins between groups were screened.

Results

The neurological impairment scores after 72 h of treatment: compared with the sham operation group, the scores of the model group, the control point group and the acupoint group were significantly increased (P<0.01); compared with the model group, the scores of the acupoint group and the control point group were significantly decreased (P<0.01, P<0.05); the score of the acupoint group was better than that of the control point group (P<0.05). The results of the protein microarray: compared with the sham operation group, 48 proteins showed up-regulated phosphorylation (≥1.5 times) in the model group and the down-regulated was 28; compared with the model group, 35 proteins showed up-regulated phosphorylation in the control point group, and the down-regulated was 24. There were 29 proteins showing up-regulated phosphorylation in the acupoint group and the down-regulated was 51. The numbers of proteins involved in the function and signal transduction pathways were also different.

Conclusion

Acupuncture at Dazhui (GV 14), Baihui (GV 20) and Shuigou (GV 25) can effectively repair brain injury. The ischemic injury of brain tissue may be caused by imbalance of a variety of proteins, and acupuncture can promote brain tissue repair by multi-functional and multi-channel regulation of the protein disorders.