蕲蛇酶减轻大鼠局灶性脑缺血再灌注损伤的实验研究(Ⅱ)

目的 从病理形态学观察蕲蛇酶对大鼠局灶性脑缺血再灌注损伤的保护作用并探讨其机制。方法 采用线栓法制备大鼠大脑中动脉栓塞后再灌注模型，光镜及电镜观察缺血3h恢复血流再灌24h后脑组织形态学变化。TTC染色观察脑梗死面积，免疫组织化学方法检测脑组织碱性成纤维细胞生长因子（bFGF）的表达。用药组分别在缺血即刻或再灌即刻静脉给予不同剂量的蕲蛇酶，观察比较模型组和蕲蛇酶所有给药组之间上述指标的变化。结果 缺血即刻给蕲蛇酶4U／kg组较再灌即刻给蕲蛇酶同剂量组显著减小梗死灶（P〈0．001），减轻脑组织病理改变，并使脑组织中bFGF蛋白表达明显增多。结论 蕲蛇酶对脑缺血再灌注损伤的神经保护作用可能与脑组织中bFGF表达增高有关；提示该药早期使用更为有效。     

运动神经元病患者胸锁乳突肌肌电图的特征

目的探讨运动神经元病(MND)患者胸锁乳突肌(SCM)肌电图的特征。方法回顾性分析461例MND患者及349例非MND患者的临床和肌电图资料。结果MND组SCM肌电图异常率(60.3%)显著高于非MND组(4.6%)(P<0.01);确诊级MND患者SCM肌电图的异常率(77.4%)明显高于其他诊断级(均P<0.01);MND组中,SCM肌电图的异常率(60.3%)低于上、下肢体肌肉的肌电图(93.2%、84.4%)(均P<0.001);MND患者SCM肌电图异常以自发电位(42.5%)和轻收缩时运动单位电位时限增宽(43.2%)最为常见;MND组有延髓症状者SCM肌电图的异常率(71.7%)明显高于无延髓症状者(54.3%)(P<0.05)。结论MND患者SCM肌电图异常率及其特异性高,为延髓肌受累的指征,有助于MND的诊断与鉴别诊断。     

The herbal preparation STW5 (lberogast®) has potent and region-specific effects on gastric motility

Functional dyspepsia (FD) is amongst the most common functional gastrointestinal disorders. Symptomatic treatment includes the use of herbal preparations whose effects on gastric motility are unclear. The present study aimed at investigating the effects of STW 5 (Iberogast), a fixed combination of hydroethanolic herbal extracts, on gastric motility in vitro. Muscle strips from guinea-pig gastric fundus, corpus and antrum were set up in organ baths either in circular or longitudinal orientation. Addition of ethanol-free STW 5 to the organ baths (32-512 microg mL(-1)) dose-dependently evoked a sustained and reversible relaxation of circular and longitudinal fundus and corpus muscle strips without changes in phasic activity. In contrast, antral muscle strips responded to STW 5 with a significant increase in the contractile force of phasic contractions without changes in tone. All effects were resistant to tetrodotoxin (0.5 micromol L(-1)), atropine (1 micromol L(-1)), omega-conotoxin GVIA (0.5 micromol L(-1)), capsaicin (1 micromol L(-1)) or L-NAME (100 micromol L(-1)), suggesting that neither nerves nor nitric oxide pathways were involved. These data demonstrate that STW 5 profoundly alters gastric motility in a region-specific but not layer-specific manner and thus implicates Iberogast in the treatment of FD patients suffering from motility disorders with impaired fundus accommodation and/or antral hypomotility.     

氯胺酮对脂多糖诱导下人脐静脉内皮细胞活化的影响

目的 观察氯胺酮和N-甲基-D-天门冬氨酸(NMDA)受体非竞争性拮抗剂MK-801对脂多糖(LPS)刺激下人脐静脉内皮细胞(HUVECs)胞间粘附分子-1(ICAM-1)表达及核因子-kappa B(NF-kB)易位表达的影响。方法 采用Jaffe方法培养的HUVECs随机分为10组:对照组(C组,RPMI-1640),LPS组(L组,LPS1μg/ml),氯胺酮组(K组,依浓度不同分为KⅠ、KⅡ、KⅢ、KⅣ亚组,即氯胺酮12.5、25.0、100、300μmol/L LPS1μg/ml),MK-801组(M组,依浓度不同分为MⅠ、MⅡ、MⅢ、MⅣ亚组,即MK-801 1.25、2.50、10、30μmol/L LPS1μg/ml)。在37℃、5％CO_2中孵育18h后,用流式细胞仪检测ICAM-1的表达阳性率。NF-kB易位表达的测定分组处理同上,在LPS1μg/ml刺激2h后,用免疫组化(SP)方法测定内皮细胞中NF-kB p65亚基的表达。结果 KⅡ、KⅢ、KⅣ亚组可抑制LPS作用下HUVECs表面ICAM-1的表达和细胞内部NF-kB的易位表达(P<0.05),且两者的变化呈正相关(r=0.985,P<0.01)。M组各亚组对LPS作用后HUVECs表面ICAM-1的表达和NF-kB的易位表达无明显影响(P>0.05)。结论 氯胺酮对炎症反应中内皮细胞的活化具有抑制作用,但并非通过NMDA受体途径。     

Effects of mycophenolic acid on IL-6 expression of human renal proximal and distal tubular cells in vitro.

BACKGROUND: Interleukin-6 (IL-6) is a multifunctional cytokine which regulates immune responses and host defence mechanisms. IL-6 has been found to be increased in certain inflammatory conditions of the kidney, in which tubular epithelial cells play a pivotal role. Human renal tubular cells express IL-6. Until now no data about the effect of the immunosuppressant drug mycophenolic acid (MPA) on IL-6 expression were available. METHODS: Proximal and distal tubular epithelial cells (PTC/DTC) have been isolated immunomagnetically. Confluent monolayers were stimulated with interleukin-1beta (IL-1beta; 25 U/ml), IL-1beta+ MPA (0.25-50 micro M) or MPA alone for 48 h. Release of IL-6 protein into the supernatant was evaluated with an enzyme immunoassay, IL-6 mRNA expression was evaluated using the Quantikine mRNA kit. RESULTS: After IL-1beta stimulation, a highly significant 2.6- (PTC) and 3.8-fold (DTC) upregulation of IL-6 expression was detectable. IL-6 mRNA was upregulated by IL-1beta [1.57- (PTC) and 2.03-fold (DTC)]. MPA inhibited this cytokine-induced IL-6 expression in a dose-dependent manner. Incubation with the lowest MPA concentration had no effect on the stimulated upregulation, whereas all higher doses significantly decreased IL-6 expression. Dexamethasone significantly inhibited the cytokine-induced IL-6 protein release in PTC, but not in DTC. CONCLUSIONS: In this study we demonstrated for the first time an inhibitory effect of MPA on the stimulated IL-6 expression of renal tubular epithelial cells. In contrast to older data, which showed a synergistic upregulation of the expression of a CC-chemokine by a combination of cytokines and MPA, in the present study we could demonstrate an immunosuppressive effect of MPA on the expression of an important cytokine.     

乳猪肝细胞的分离、培养及功能检测

目的 为构建生物人工肝进行肝细胞的准备。方法 采用胶原酶半原位灌流法分离单个乳猪肝细胞,并对其活力及单层和聚集培养后的白蛋白、尿素合成功能进行检测。结果 采用本方法从每头乳猪中分离到的单个肝细胞数为(3.1±1.5)×10~(10),活性超过95％。在加入激素和生长因子的培养基中单层培养时,肝细胞功能良好,可维持2周左右。而在未加入激素和生长因子的培养中肝细胞虽能存活1周,但功能于24h后即丧失。球形聚集培养可实现肝细胞的大量培养,且生物学活性较单层培养显著提高。结论 采用胶原酶半原位灌注法所得单个乳猪肝细胞基本能满足构建生物人工肝对肝细胞数量的要求。聚集培养接种密度大,细胞生物学活性高,可用于构建生物人工肝。     

Rosiglitazone down-regulates lipopolysaccharide-induced expression of CD40 and intercellular adhesion molecule 1 in rat peritoneal mesothelial cells through a NF-κB dependent mechanism

Objective To investigate the effect and mechanism by which PPARγ ligand, rosiglitasone, regulates the expression of CD40 and intercellular adhesion molecule 1 (ICAM-1) in the rat peritoneal mesothelial cells (RPMCs) induced by lipopolysaccharide (LPS). Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were randomly divided into groups as follows: medium, LPS (5 mg/L), LPS (5 mg/L)+BAY11-7085(5 μmol/L, NF-κB inhibitor), rosiglitazone (10 μmol/L or 20 μmol/L, peroxisome proliferator-activated receptor γ activator), LPS (5 mg/L)+rosiglitazone (10 μmol/L)+GW9662 (3 μmol/L, peroxisome proliferator-aetivatcd receptor γ antagonist), and LPS (5 mg/L)+vehicle (DMSO 0.2 ml/L). The expressions of CD40 and ICAM-1 RNA in RPMCs were examined by RT-PCR after 3 hour treatment, and the protein expressions of CD40, ICAM-1, p-NF-κB p65 and p-IκBα were examined by Western blot or immunofluorescence after 24 hour treatment. Results Following treatment with LPS, both the expressions of CD40 and ICAM-1 protein in RPMCs were up-regulated significantly (P<0.05), and the phosphoralation of p65 was increased greatly (1.10±0.17 vs 0.55±0.06, P<0.05). BAY11-7085 (5 μmol/L) significantly decreased the protein expression of p-p65 (0.22±0.11 vs 1.10±0.17, P<0.01), CD40 (0.34±0.02 vs 0.50±0.06, P<0.05) and ICAM-1 (0.35±0.16 vs 0.74±0.03, P<0.05). Pretreated with rosiglitazone for 3 h then added with LPS for 1 h, the levels of p-p65, CD40 and ICAM-1 in RPMCs were significantly decreased compared with those of LPS group (0.77±0.08 vs 0.90±0.10, P相似文献   

Pycnogenol® as an Adjunct in the Management of Childhood Asthma

A randomized, placebo-controlled, double-blind study involving 60 subjects, aged 6-18 years old, was conducted over a period of 3 months to determine the effect of Pycnogenol® (a proprietary mixture of water-soluble bioflavonoids extracted from French maritime pine) on mild-to-moderate asthma. After baseline evaluation, subjects were randomized into two groups to receive either Pycnogenol® or placebo. Subjects were instructed to record their peak expiratory flow with an Assess® Peak Flow Meter each evening. At the same time, symptoms, daily use of rescue inhalers (albuterol), and any changes in oral medications were also recorded. Urine samples were obtained from the subjects at the end of the run-in period, and at 1-, 2-, and 3-month visits. Urinary leukotriene C₄/D₄/E₄ was measured by an enzyme immunoassay. Compared with subjects taking placebo, the group who took Pycnogenol® had significantly more improvement in pulmonary functions and asthma symptoms. The Pycnogenol® group was able to reduce or discontinue their use of rescue inhalers more often than the placebo group. There was also a significant reduction of urinary leukotrienes in the Pycnogenol® group. The results of this study demonstrate the efficacy of Pycnogenol® as an adjunct in the management of mild-to-moderate childhood asthma.     

再生障碍性贫血造血细胞Fas抗原表达与T淋巴细胞功能的研究

目的 研究造血细胞凋亡与T淋巴细胞免疫在再生障碍性贫血(再障，Aplastic anemia，AA)发病机制中的作用及两者相关性。方法 采用流式细胞仪测定40例AA患者和15例非血液、免疫系统疾病对照者骨髓单个核细胞(BMMNC)的总CD34^ 、CD34^ Fas^ 、CD34^ Fas^-、CD3^ 、CD8^ 、CD3^ 、CD3^ CD25^ 标记值。结果 (1)与对照组比较，AA组总CD34^ 细胞％明显减少而其Fas表达率(以占总CD34^ 细胞％为计)明显增高。(2)AA组CD34^ 细胞％数与其Fas抗原表达率无明显负相关。(3)AA组T细胞％明显增多，且以CD8^ 细胞和CD3^ CD25^ 细胞增多为主。(4)AA组CD34^ 细胞％数与其T细胞活化状态无显著负相关。(5)AA组CD34^ 细胞Fas表达率与其T细胞活化状态无显著正相关。结论 AA骨髓存在着造血细胞数量减少和T淋巴细胞亚群数量、功能的异常，造血细胞数量减少还可能与Fas以外途径诱导的凋亡过度有关。骨髓造血细胞凋亡过度可能有活化T淋巴细胞免疫以外的途径诱导Fas途径或活化T淋巴细胞可以通过Fas之外的途径诱导造血细胞凋亡。