层层静电自组装多聚电解质共混膜与组织工程心脏瓣膜细胞的黏附

背景:组织工程心脏瓣膜研究中的核心和难点是种子细胞在支架材料上的黏附.层层静电自组装技术能够将多聚阴阳离子组装成多层膜状结构增加材料表面的生物相容性.目的:拟对猪去细胞瓣膜在体外进行层层静电自组装成多聚电解质共混膜,并且检测共混膜列种子细胞黏附的效果.设计时间及地点:细胞学体外观察性实验,于2008-09/2009 02在解放军第二军医大学长海医院胸心外科实验室和复旦大学高分子科学系实验室完成.材料:新鲜猪心脏瓣膜取自上海五丰上食肉联厂,采用酶消化法将猪主动脉瓣叶行脱细胞处理.密度梯度离心法从骨髓中获取骨髓单个核细胞,体外扩增培养后鉴定.方法:通过层层静电自组装技术构建20,10,5,0层的壳聚糖,透明质酸共混膜履盖在猪去细胞瓣膜支架上.将骨髓基质干细胞在涂膜支架上种植培养并检测支架对种子细胞的黏附效果.主要观察指标:扫描电镜观察、细胞计数、MTT检测组织工程心脏瓣膜上种子细胞的形态和细胞黏附效果.结果:扫描电镜观察20层壳聚糖,透明质酸多层膜样品上细胞生长情况比较好,样品上可以铺满细胞.而在组装层数为5层和10层的样品上,细胞生长的比较少.构建层数为20层的瓣叶,其细胞计数及吸光值高于10层和5层构建的瓣叶(P<0.01).构建层数为10层和5层的瓣叶与0层比较差异无显著性意义(P>0.05).结论:通过层层静电自组装技术构建的共混膜能够促进种子细胞的黏附,而且有一定的量效关系.     

隔药灸对慢性炎性内脏痛大鼠下丘脑SP、5-HT及c-Fos的影响

目的：本研究主要观察隔药灸对慢性炎性内脏痛大鼠下丘脑P 物质、5-羟色胺和c-Fos含量的影响,探讨隔药灸治疗慢性炎性内脏痛的镇痛机制。方法：雄性SD大鼠随机分为正常组、模型组、隔药灸组和假隔药灸组。采用三硝基苯磺酸合50%乙醇灌肠的方法制备慢性炎性内脏痛大鼠模型。隔药灸组选取双侧天枢穴、气海穴进行隔药饼灸治疗;假隔药灸组仅在穴位上放置药饼和艾炷,不点燃艾炷;模型组和正常组均不进行治疗,只做与隔药灸组相同的固定。治疗结束后,采用腹壁撤回反射评分（AWR）、机械性缩足反射阈值（MWT）、热缩足潜伏期（TWL）观察大鼠痛行为的变化,采用ELISA技术检测各组大鼠下丘脑P物质、5-羟色胺、c-Fos的含量。结果：与正常组比较,模型组大鼠AWR评分显著升高（P<0.05）,MWT、TWL评分均显著降低（P<0.01）;与模型组、假隔药灸组比较,隔药灸组大鼠AWR评分降低（P<0.05）,MWT、TWL 评分均显著升高（P<0.01）;假隔药灸组与模型组比较,AWR、MWT、TWL评分差异均无统计学意义（P>0.05）。与正常组比较,模型组大鼠下丘脑P物质、5-羟色胺、c-Fos含量均显著升高（P<0.01）;与模型组、假隔药灸组比较,隔药灸组大鼠下丘脑P物质、5-羟色胺、c-Fos含量均显著降低（P<0.01）;假隔药灸组与模型组比较,下丘脑P 物质、5-羟色胺、c-Fos含量差异均无统计学意义（P>0.05）。结论：隔药灸能降低慢性炎性内脏痛大鼠下丘脑P物质、5-羟色胺、c-Fos的含量,该作用可能是隔药灸治疗慢性炎性内脏痛发挥镇痛作用的重要中枢机制。     

艾灸对慢性萎缩性胃炎大鼠外周血基因表达谱的影响

目的 探讨隔药饼灸对慢性萎缩性胃炎大鼠外周血基因表达谱的影响，为后续研究提供研究方向。方法 将雄性Wistar大鼠随机分成正常组、模型组、隔药饼灸组、电针组和西药组，均采用MNNG诱导CAG大鼠模型，取穴中脘、气海，隔药饼灸每次每穴1壮，每日1次，共4周；电针频率100 Hz，强度1-3 mA，20 min，每日1次，共4周；西药采用叶酸悬浊液1.0 mL/100 g灌胃，每天1次，持续4周。采用Trizol法提取各组CAG大鼠的外周血总RNA，质控后建立文库，采用RNA-seq高通量测序，结合生物信息学的方法深入分析隔药饼灸逆转的差异表达基因。结果 与正常组比较，CAG组大鼠外周血有1 542个差异表达基因，涉及的通路主要有癌症、免疫疾病、能量代谢、消化系统、免疫系统通路。隔药饼灸干预CAG大鼠的外周血差异表达基因2 956个，涉及的通路与分子相互作用、信号转导、癌症、免疫疾病、氨基酸代谢、能量代谢、脂质代谢、维生素代谢、核苷酸代谢、消化系统、免疫系统、神经内分泌系统等相关；电针干预CAG外周血的差异表达基因3 136个，涉及的通路与癌症、免疫疾病、氨基酸代谢、能量代谢、脂质代谢、维生素代谢、核苷酸代谢、消化系统、免疫系统、神经内分泌系统等相关；西药干预CAG外周血差异表达基因1 308个，涉及的通路与蛋白翻译、信号转导、癌症、免疫系统、消化系统等相关。经RT-qPCR验证，各组外周血Foxo3、Uba52、S100a1、Nod2的结果和RNA-seq测序结果一致。结论 隔药饼灸、电针和西药干预CAG的外周血差异表达基因的数量不同，西药较针灸改变CAG的差异基因数目少。3种干预方法均能影响CAG大鼠外周血免疫相关、肿瘤相关的基因表达谱，隔药饼灸和电针干预的差异基因还涉及能量、氨基酸、脂质、维生素代谢。     

针刺对阻塞性睡眠呼吸暂停低通气综合征患者深浅睡眠的调节作用

目的:观察针刺对阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者深浅睡眠结构及相关因素的影响。方法:30例OSAHS患者,选取上廉泉、哑门、风池、迎香、列缺、隐白、照海等穴位,每次留针30min,每周治疗3～5次,10次为1个疗程,治疗3个疗程。分析治疗前后脑电图的睡眠相关指标及嗜睡评估。结果:与治疗前相比,针刺后浅睡眠%、深睡眠%有明显的改善,统计学分析显示有非常显著性差异(P0.01)。结论:针刺治疗可使浅睡眠比例减少,增加深睡眠比例,调整睡眠结构,提高睡眠质量。     

基于《中医针灸信息库》对针灸治疗抑郁症多因素分析

目的回顾2004年至2013年针灸文献,探索提高针灸治疗抑郁症的临床疗效。方法根据《中医针灸信息库》,采用计量分析、单因素方差分析和t检验对针灸治疗抑郁症的常用穴位、常用治疗方法、治疗次数、HAMD值、疾病程度、不同病因进行多因素分析。结果经多因素分析证实中度抑郁症是针灸治疗的适宜证;治疗次数以11~20次为宜;百会、内关应用频次为最高,百会配内关、太冲,百会配内关、三阴交可作为最佳配穴;治疗方法以针刺为主,单纯电针疗法和单纯针灸疗法临床疗效优于单纯氟西汀,针灸配合氟西汀可提高临床疗效。结论针灸治疗抑郁症有较好的疗效。     

隔药灸和电针对大鼠克罗恩病肠纤维化血清学指标的影响

目的:观察隔药灸和电针对克罗恩病(Crohn's Disease,CD)肠纤维化大鼠血清透明质酸(Hyaluronic Acid,HA)、Ⅲ型前胶原(Procollagen Ⅲ,PC Ⅲ)、Ⅳ型胶原(Collagen Ⅳ,C Ⅳ)的影响,探讨针灸抗CD大鼠肠纤维化的作用与机制.方法:SD大鼠随机分为正常组、模型组、隔药灸组、电针组和药物组.采用三硝基苯磺酸制备大鼠CD模型,应用Masson染色光镜下观察结肠胶原纤维增生情况,采用放射免疫分析技术检测大鼠血清HA、PC Ⅲ、CⅣ含量.结果:与正常组比较,模型组大鼠结肠组织胶原纤维增生,血清HA、PCⅢ、CⅣ含量增加;与模型组比较,隔药灸组、电针组及西药组胶原纤维表达减少,血清中HA、PCⅢ、CⅣ含量降低,与电针组、西药组比较,隔药灸组胶原纤维表达减少,血清CⅣ含量降低.结论:隔药灸和电针均可改善CD大鼠肠纤维化病理状态,降低血清HA、PCⅢ、CⅣ含量.

Abstract：

Objective:To observe the impact of herb-partitioned moxibustion and electroacupuncture on hyaluronic acid(HA),procollagen Ⅲ(PC Ⅲ)and collagen Ⅳ(CⅣ)of intestinal fibrosis in rats with Crohn's disease(CD),and discuss the effect and the mechanism of acupuncture on treating intestinal fibrosis of rats.Methods:SD rats were randomly divided into control group,model group,herb-partitioned moxibustion group,electroacupuncture group and medicine group.The model of CD was developed by trinitrobenzene sulfonic acid.Masson stain was used to observe the collagen fibers hyperplasia in colon and radioimmunoassay was adopted to detect the content of HA,PC Ⅲ and C Ⅳ in serum.Results:Compared with the control group,the proliferation of colonic collagen fibers,as well as the HA,PC Ⅲ and CⅣ in serum increased in the model group.Compared with the model group,the expression of collagen fibers,serum HA,PC Ⅲ and C Ⅳ decreased in the herb-partitioned moxibustion group,electroacupuncture group and medicine group.The expression of collagen fibers serum HA,PC Ⅲ and C Ⅳ in the herb-partitioned moxibustion group were lower than in the electroacupuncture group and medicine group.Conclusion:Herb-partitioned moxibustion and electroacupuncture effectively improved the pathological state of intestinal fibrosis in rats with CD and reduced the content of HA,PC Ⅲ and C Ⅳ in serum.     

Clinical observation of Zhen’ai needling method in <Emphasis Type="Italic">Nei Jing</Emphasis> (<Emphasis Type="Italic">Classic of Internal Medicine</Emphasis>) on improving quality of life in patients with allergic rhinitis

Objective

To observe the effect of Zhen’ai needling method in Nei Jing (Classic of Internal Medicine) on quality of life in patients with allergic rhinitis (AR).

Methods

A total of 99 patients with AR who met the inclusion criteria were randomized into a conventional needling group (51 cases) and a Zhen’ai needling group (48 cases). The conventional needling group was treated with acupuncture at Zusanli (ST 36), Hegu (LI 4), Juliao (ST 3), Yingxiang (LI 20), Yintang (GV 29), Shangxing (GV 23) and Baihui (GV 20). The Zhen’ai needling group added acupoints of Zhen’ai needling method {Lieque (LU 7), Shanglianquan [Extra, located at 1 cun above Lianquan (CV 23)] and Tianrong (SI 17)} in addition to points in the conventional needling group. The needles were retained for 30 min. The treatment was performed twice a week. All patients were assessed by total nasal symptom score (TNSS), total ocular symptom score (TOSS) and sino-nasal outcome test-20 (SNOT-20) before and after 15 treatments to evaluate the quality of life. The clinical efficacy was evaluated after 15 treatments.

Results

During the treatment, 8 cases dropped out in the conventional needling group and 7 cases in the Zhen’ai needling group. There were no significant differences in TNSS, TOSS and SNOT-20 scores between the two groups before treatment (all P>0.05). After treatment, the total effective rate of the Zhen’ai needling group was 92.7%, versus 88.4% in the conventional needling group. There was no significant difference between the two groups (P>0.05). After treatment, there were no significant differences between the two groups in TNSS and TOSS scores (both P>0.05). The SNOT-20 score of the Zhen’ai needling group was lower than that of the conventional needling group, and the difference was statistically significant (P<0.05). There were statistical differences between the two groups in the scores of cough and nasal reflux (to the throat) of the NOT-20 scale (both P<0.05).

Conclusion

Both Zhen’ai needling method and conventional acupuncture are effective in the treatment of AR. Zhen’ai needling method has a better effect than conventional acupuncture in improving cough and nasal reflux (to the throat) in AR patients.

     

艾灸对克罗恩病大鼠结肠NF-κB p65及TNF-α、IL-1β调节作用的研究

目的：通过观察隔药灸对克罗恩病大鼠结肠NF-κB p65及炎症因子TNF-α、IL-1β表达的影响,探讨艾灸治疗克罗恩病的效应机制。方法：清洁级雄性SD大鼠随机分为4组,分别为正常组、模型组、隔药灸组和西药组,采用三硝基苯磺酸（TN-BS）合乙醇溶液灌肠制备克罗恩病大鼠模型,模型制备成功后,隔药灸组取天枢穴（双）、气海穴进行隔药灸治疗,西药组采用美沙拉秦灌胃治疗。治疗结束后,采用HE染色,光镜下观察各组大鼠结肠组织病理学变化,应用免疫组织化学技术观察各组大鼠结肠NF-κB p65与TNF-α、IL-1β的表达,并进行图像分析。结果：与正常组比较,模型组大鼠结肠组织呈现裂隙状溃疡及炎症表现,部分大鼠结肠在黏膜下层可见肉芽肿形成,结肠NF-κB p65、TNF-α、IL-1β的表达均显著增高（均P〈0.01）;与模型组比较,隔药灸组、西药组大鼠结肠形态结构改善,肠道炎症减轻,结肠NF-κB p65与TNF-α、IL-1β的表达均显著降低（均P〈0.01）;相关性分析结果显示,TNF-α、IL-1β的表达与NF-κB p65的表达均呈正相关性。结论：隔药灸能下调克罗恩病大鼠结肠NF-κB p65、TNF-α、IL-1β的表达;艾灸（隔药灸）可能通过抑制克罗恩病大鼠结肠NF-κB p65的表达,进而减少其下游炎症因子TNF-α、IL-1β的表达,从而减轻肠道炎症,改善结肠组织形态结构,发挥治疗作用。     

Influence of electroacupuncture on histomorphology of lacrimal glands and ocular surface in experimental dry eye syndrome

Objective

To observe the influence of electroacupuncture (EA) on histomorphologies of lacrimal glands, cornea and conjunctiva in experimental dry eye syndrome, and to explore the repair effects of EA on lacrimal glands and ocular surface damage.

Methods

Twenty-four healthy male New Zealand rabbits were randomly divided into a normal group, a model group, an EA group and a medication group, 6 rabbits in each group. Experimental dry eye syndrome models were prepared in rabbits by using 0.1% benzalkonium chloride for eye drops. Tear secretion volume, break-up time of tear film (BUT) and corneal fluorescein staining score were observed before and after the treatment. Periodic acid Schiff (PAS) staining method was used to observe the changes of conjunctival goblet cells in rabbits. After hematoxylin eosin (HE) staining, morphological changes of rabbit cornea, conjunctiva and lacrimal gland tissues were observed under light microscope.

Results

Compared with the normal group, tear secretion volume and BUT were significantly reduced (both P<0.01), while the corneal fluorescein staining score was significantly increased (P<0.01) in the model group. Compared with the model group, tear secretion volume and BUT were significantly increased, while the corneal fluorescein staining score was significantly decreased in the EA group and the medication group (all P<0.01). Compared with the normal group, the number of conjunctival goblet cells in the model group was significantly reduced; compared with the model group, the numbers of conjunctival goblet cells were all relatively higher in the EA group and the medication group. Pathological lesions of cornea, conjunctiva and lacrimal glands all showed improvement by HE staining in the EA group and the medication group after the intervention.

Conclusion

EA can improve tear secretion and tear film stability of rabbit dry eye syndrome, and repair the pathologic lesions of conjunctival goblet cells, corneal epithelia, cornea, conjunctiva and lacrimal glands.

     

Clinical Research of Rolling Manipulation in Treating Knee Osteoarthritis

Knee Osteoarthritis （KOA） is also called degenerative osteoarthritis. With its incidence reaching 13.2 % in men and 28.3 % in women, it has become the most common joint disease and also one of the major reasons for old people to be disabled. The feature of the disease is progressive degeneration in cartilage tissue of the joint, osteophyma formed in the border of the joint and reactive change in the sclerotin of the cartilage. The patients can experience pain and weakness of the knee joint, difficulty in walking, standing and climbing stairs, which severely disturb the daily life of the patients.