人参皂甙Rg1对炎性痛大鼠吗啡耐受时脊髓背角细胞凋亡的影响

目的 探讨人参皂甙Rg1对炎性痛大鼠吗啡耐受时脊髓背角细胞凋亡的影响.方法 雄性SD大鼠,体重280～320 g,采用右踝关节腔内注射完全弗氏佐剂的方法制备炎性痛模型.取模型制备成功的24只大鼠,采用随机数字表法,将其随机分为4组(n=6):生理盐水对照组(C组)、吗啡耐受组(M组)、人参皂甙Rg1组(G组)及吗啡复合人参皂甙Rg1组(MG组).致炎后3d时M组、G组及MG组分别鞘内注射吗啡10 μg、人参皂甙Rg1 100μg及吗啡10μg+人参皂甙Rg1 100 μg,C组给予等容量生理盐水,吗啡2次/d,人参皂甙Rg1 1次/d,连续7d.分别于致炎前(T1)、鞘内给药前1d(T2)、给药1、3、5、7 d(T3-6)时测定机械缩足阈值(PWT).最后一次测定痛阈后处死大鼠,取L3-5节段脊髓组织,采用TUNEL染色法测定细胞凋亡情况,计算脊髓背角细胞凋亡率.结果 与T1时比较,4组T2-6时PWT降低(P＜0.01);与T2时比较,M组T3.4时PWT升高,G组和MG组T3～6时PWT升高(P＜0.05);与C组比较,M组和MG组PWT和脊髓背角细胞凋亡率升高(P＜0.05),G组上述指标差异无统计学意义(P＞0.05);与M组比较,MG组PWT升高,脊髓背角细胞凋亡率降低(P＜0.05).结论 人参皂甙Rg1预防吗啡耐受形成的机制与降低炎性痛大鼠脊髓背角细胞凋亡有关.     

The effects of dopamine receptor agonists on BK Ca channels and signal transduction mechanism in corpus cavernosal smooth muscle cells

In this study, we investigated the effect of dopamine receptor agonists on potassium channels' activity and their signal transduction pathway in corporal smooth muscle cells. We used cultured human corporal smooth muscle cells. The whole cell and cell-attached configuration of the patch-clamp technique were used for electrophysiological recordings, and enzyme immunoassay was used for measuring cyclic AMP (cAMP) and cyclic GMP levels. Extracellular application of 10 microM dopamine and apomorphine significantly increased whole-cell K(+) currents by 283.5+/-55.7% (at +60 mV; n=12, P<0.001), 292.4+/-58.8.0% (at +60 mV; n=9, P<0.005), respectively. We confirmed that the increase in whole-cell currents was mainly due to activation of the tetraethylammonium-sensitive large conductance Ca(2+)-activated K(+) channels (BK(Ca) channels). Enzyme immunoassay indicated that dopamine and apomorphine stimulates cAMP levels in corporal smooth muscle cells in a concentration-dependent fashion. The activation of BK(Ca) channels by dopamine receptor agonists in corporal smooth muscle cells might be one of the mechanisms in inducing penile erection.     

术后采用人参皂甙Rg3联合丝裂霉素加呋喃尿嘧啶方案对进展期胃癌的疗效

目的探讨人参皂甙Rg3(Rg3)联合丝裂霉素加呋喃尿嘧啶(MF)化疗方案(Rg3 MF)对进展期胃癌术后的治疗效果。方法将71例进展期胃癌术后患者随机分为对照组(33例)和观察组(38例),前者用MF方案,后者用Rg3加MF方案治疗。检测血清血管内皮生长因子(VEGF)水平,并对预后进行对比分析。结果进展期胃癌患者血清VEGF含量[(297.8±129.6) pg/ml]明显高于正常组[(212.3±67.5)pg/ml](P<0.01),且与胃癌患者肿瘤的浸润深度、淋巴结转移、肿瘤大于4cm及TMN分期有关(均P<0.05)。术后14周时检测血清VEGF水平,观察组已明显低于术前(P<0.05)、而接近正常组水平,对照组仅降至术前水平。观察组、对照组中位生存时间分别为40和25个月。观察组的术后累计生存率高于对照组(P=0.047)。结论进展期胃癌患者术后采用MF方案联合Rg3治疗可明显降低血清VEGF含量,提高生存率。     

Effects of nitric oxide on the Ca2+-activated potassium channels in smooth muscle cells of the human corpus cavernosum

Relaxation of the corpus cavernosum smooth muscle is an absolute prerequisite of penile erection. Potassium channels play a role in the physiologic regulation of corporal smooth muscle tone. Among the several subtypes of potassium channels, Ca2 +-activated potassium channel (KCa channel) subtypes are thought to be the most physiologically relevant in the regulation of corporal smooth muscle tone. The purpose of this study was to investigate the effects of nitric oxide (NO) and sildenafil on the KCa channels and elucidate the mechanisms of action on the KCa channels in smooth muscle cells of the human corpus cavernosum. The conventional patch-clamp technique was applied to short-term cultured smooth muscle cells of the human corpus cavernosum. Single-channel currents were recorded in cell-attached or inside-out patches, and whole-cell currents were recorded in perforated-patches. In cell-attached patches, sildenafil alone did not activate the KCa channels but sildenafil enhanced the NO-induced activation of KCa channels. The open probability of KCa channels was increased significantly after application of NO donor, SIN-1 (100 microM) (47 +/- 7.1%, n = 10, P=0.002). The application of sildenafil (100 nM) with SIN-1 (100 microM) markedly increased the open probability of KCa channels (148 +/- 24%, n = 8, P < 0.001). The activation by SIN-1 or sildenafil with SIN-1 was completely blocked by pretreatment of the soluble guanylate cyclase inhibitor, ODQ (10 microM). In inside-out patches. SIN-1 or sildenafil with SIN-1 failed to activate KCa channels at pCa 7.5 (n=5). SIN-1 increased the whole cell outward K+ currents in all holding potential. The increased IK by SIN-1 was inhibited by charybdotoxin (CTX) about 70%. These data provide compelling evidence consistent with the involvement of the KCa channel subtype in modulating NO-induced relaxation responses in human corporal smooth muscle. Furthermore, the activation of KCa channels is thought to be mediated by activation of soluble guanylate cyclase, leading to increased intracellular levels of cyclic GMP and the subsequent activation of protein kinase rather than direct NO effect.     

人参皂苷Rg3对人前列腺癌细胞EphB4及抗凋亡蛋白bcl-xl表达的影响

目的：研究人参皂苷Rg3对人前列腺癌细胞株PC-3中EphB4及抗凋亡蛋白bcl—xl表达的影响。方法：用浓度为0、5、10、20和40μmol／L的人参皂苷Rg3处理PC-3细胞24h,然后采用四甲基偶氮唑盐（MTT）方法检测人参皂苷Rg3对PC-3细胞增殖的抑制作用,用倒置显微镜和流式细胞术观察人参皂苷Rg3对PC-3细胞凋亡的诱导作用,用RT—PCR和Western blot方法检测经不同浓度人参皂苷Rg3处理后PC-3细胞中EphB4和bcl-x1的表达情况。结果：5、10、20和40μmol／L的人参皂苷Rg3对PC3细胞增殖的抑制率分别为20．93％、31．32％、51．63％、65．43％。5～40μmol／L的人参皂苷Rg3处理细胞呈明显的凋亡形态学改变,40umol／L人参皂苷Rg3处理PC-3细胞24h后,凋亡细胞占（12．10±1．2）％,处理组比正常对照组（3．18±2．1）％凋亡明显增加,差异有统计学意义（P〈0．05）。结论：不同浓度人参皂苷Rg3处理PC-3细胞24后,EphB4的表达随人参皂苷Rg3浓度的增加而逐渐减弱,而且bcl—xl的表达随人参皂苷Rg3浓度的增加逐渐减弱。     

急性胆囊炎腹腔镜手术时机的选择

目的探讨急性胆囊炎腹腔镜胆囊切除术（laparoscopic cholecystectomy,LC）的手术时机。方法回顾分析2006年7月～2011年3月186例急性胆囊炎行LC的临床资料,根据病程分为3组：病程72 h内为组1（n=81）,病程72 h～1周为组2（n=67）,病程1～2周为组3（n=38）;比较3组中转率、手术时间、术中出血量等。采用四孔法,前后结合解剖Calot三角,顺逆结合切除胆囊,并遵循疑难复杂胆囊结石LC的其他基本原则。结果 LC成功158例,中转开腹28例,中转率15.1%（28/158）。无手术并发症,无死亡病例。组1中转开腹率6.2%（5/81）显著低于组3 36.8%（14/38）（χ2=18.133,P=0.000）,但与组2 13.4%（9/67）无统计学差异（χ2=2.257,P=0.133）;组2中转开腹率13.4%（9/67）显著低于组3 36.8%（14/38）（χ2=7.768,P=0.005）。组1手术时间（42.6±11.4）min显著短于组2（77.4±12.6）min（q=24.863,P〈0.05）和组3（113.9±12.1）min（q=42.784,P〈0.05）,组2手术时间显著短于组3（q=21.206,P〈0.05）。组1出血量中位数20 ml（5～45 ml）显著少于组2 55 ml（30～90 ml）（Z=-6.819,P=0.000）和组3 110 ml（60～145 ml）（Z=-8.367,P=0.000）,组2出血量显著少于组3（Z=-5.306,P=0.000）。组1住院时间（6.9±2.2）d显著短于组2（11.3±2.9）d（q=14.762,P〈0.05）和组3（18.4±2.6）d（q=32.403,P〈0.05）,组2住院时间较组3显著缩短（q=19.370,P〈0.05）。结论急性胆囊炎72 h以内行LC最佳,手术时间较短,出血量较少,住院时间缩短,中转开腹率较低。     

术前输注头孢呋辛钠后不同靶控浓度丙泊酚对妇科手术患者心室复极的影响

目的观察术前预防性输注头孢呋辛钠后不同靶控浓度丙泊酚对妇科手术患者心室复极的影响。方法选择择期妇科手术患者59例,年龄18~65岁,ASAⅠ或Ⅱ级,采用随机数字表法分为三组。入室补液并静滴头孢呋辛钠2.5g(溶于100ml生理盐水中)后,分别泵注丙泊酚血浆靶控浓度为2μg/ml(P2组,n=20)、3μg/ml(P3组,n=19)、4μg/ml(P4组,n=20)。分别在抗生素输注前(T_0)、抗生素输注完毕(T_1)、及丙泊酚达到靶控浓度时(T_2),测量并计算QT间期、QTc间期、T波峰值至终点时间(T_p-e间期)及Tp-e/QT。结果与T0时比较,T_1时P4组QTc间期[(469.9±34.0)ms vs.(451.2±24.9)ms]、Tp-e间期[(107±25)ms vs.(94±20)ms]、Tp-e/QT(0.260±0.058vs.0.236±0.043)明显延长(P0.05);与T_1时比较,T_2时P2组[(437.4±24.4)ms vs.(453.3±28.0)ms]和P4组[(438.8±29.9)ms vs.(469.9±34.0)ms]QTc间期明显缩短(P0.05)。结论丙泊酚可改善头孢呋辛钠引起的心室复极不均一性。     

血清胰岛素样生长因子-1对小鼠乳腺癌组织凋亡相关基因表达的影响

目的 在肝特异性胰岛素样生长因子1(IGF-1)基因缺失小鼠(LID小鼠)及正常小鼠体内建立原发性乳腺癌模型,比较不同血清IGF-1水平下凋亡相关基因的表达情况.方法 LID小鼠及正常小鼠各50只,用化学致癌剂7,12-二甲基苯蒽诱导原发性乳腺癌;分别随机选取25只小鼠使用人参皂甙Rg3进行干预治疗.比较各组肿瘤发生情况,应用流式细胞术及基因芯片技术分析凋亡相关基因的表达情况.结果 未管饲人参皂甙Rg3的正常小鼠乳腺癌发生率为66.7%,高于其他各组(P＜0.05);而管饲人参皂甙Rg3的LID小鼠乳腺癌发生率为12.0%,低于其他各组(P＜0.05).未管饲人参皂甙Rg3的正常小鼠肿瘤细胞凋亡比例为(2.7±0.7)%,而管饲人参皂甙Rg3的LID小鼠凋亡比例为(14.0±1.7)%.基因芯片显示正常小鼠与LID小鼠相比,LTA、LTB、TNF-α、TRAIL、TRANCE、BLK、BOK、CASP8、TRAF5、APAF1等基因表达下调,LTBR、TRAF4等基因表达上调.而应用人参皂甙Rg3治疗可改变以上基因的表达情况.结论 降低血清IGF-1水平可影响一系列凋亡相关基因的表达,从而促进细胞凋亡,对乳腺肿瘤的发生发展具有抑制作用.人参皂甙Rg3对这一效应具有协同作用.     

Characterization of ATP-sensitive potassium channels in human corporal smooth muscle cells.

Potassium (K) channels play a significant role in modulating human corporal smooth muscle tone, and thus, erectile capacity. Recent pharmacological studies indicate that the metabolically-regulated K channel (KATP) may be an important modulator of human penile erection with significant therapeutic potential. The goal of these initial studies, therefore, was to utilize patch clamp techniques to characterize the putative KATP subtype(s) present in cultured and freshly isolated human corporal smooth muscle cells. In the cell-attached patch mode, two distinct unitary K+ currents were identified whose respective conductance values were similar in cultured and freshly isolated smooth muscle cells. In cultured myocytes, the measured conductance values in symmetric KCl (140 mM) solutions were 59.1 +/- 2.7 pS and 18.4 +/- 2.1 pS (n = 5 cells). Under identical experimental conditions in freshly isolated myocytes, corresponding conductance values were 59.2 +/- 3.7 pS and 18.5 +/- 2.4 pS, respectively (n = 4 cells). I-V curves constructed during step depolarization (-60 to +80 mV), revealed a linear I-V relationship for both unitary conductances. Single channel records documented that both conductances were reversibly inhibited by the application of ATP (1-3 mM) to the bath solution in the inside-out attached patch configuration. The unitary activity of both K channel subtypes was significantly increased by the application of pinacidil (10 microM) and levcromakalim (10 microM). Whole cell patch recordings documented a glibenclamide-sensitive, pinacidil- and levcromakalim-induced increase in the whole cell outward K+ current during step depolarization (-70 mV to +130 mV) of 105 +/- 37%, 139 +/- 42%, respectively. These data confirm and extend our previous observations, and provide the first evidence for the presence of KATP channel subtypes in human corporal smooth muscle cells.     

Ginsenoside Rb1 blocks Homocysteine-induced endothelial dysfunction

Background. Homocysteine (HCY) is an independent risk factor for atherosclerosis. This study is to investigate the effect of ginsenoside Rb1, a major constituent of ginseng, on HCY-induced endothelial dysfunction and molecular changes in porcine coronary arteries. Methods. Coronary arteries were harvested from pig hearts and cut into 5-mm rings, which were divided into 6 groups including control, HCY alone (50 μM), low-dosed (1 μM), or high-dose (10 μM) Rb1 alone, and HCY plus low-dose or high-dose Rb1. After 24 h incubation, the rings were analyzed for vasomotor function in response to U46619, bradykinin, and sodium nitroprusside (SNP), respectively. In addition, superoxide anion was assessed by lucigenin-enhanced chemiluminescence analysis. Endothelial nitric oxide synthase (eNOS) expression was studied by real-time RT-PCR and Western blot. Results. Endothelium-dependent relaxation (bradykinin) was significantly reduced in rings treated with HCY alone as compared to the control (49.80% versus 71.77%, n = 8, P < 0.05), while either high-dose or low-dose Rb1 alone did not affect endothelium-dependent relaxation. Low-dose Rb1-HCY combined group had partially improved endothelium-dependent relaxation (54.44%), while high-dose Rb1-HCY combined group showed complete recovery of endothelium-dependent relaxation (72.89%). There was no substantial difference in maximal contraction induced by U46619 or endothelium-independent relaxation by SNP among all groups (P > 0.05). Moreover, superoxide anion was markedly increased by 137% in HCY-treated group as compared to the control, but there was no significant changes from the control in all other groups (P > 0.05). Last, eNOS mRNA and protein levels were substantially reduced in HCY-treated group, but not in Rb1-HCY combined groups. Conclusions. This is the first study to demonstrate that ginsenoside Rb1 can effectively block HCY-induced endothelial dysfunction and superoxide anion production as well as eNOS down regulation in porcine coronary arteries. This study suggests that ginseng and its active constituents may have potential clinical applications in controlling HCY-associated vascular injuries.     

Intracavernosal injections of vascular endothelial growth factor protects endothelial dependent corpora cavernosal smooth muscle relaxation in the hypercholesterolemic rabbit: a preliminary study.

Atherosclerosis is a major risk factor for erectile dysfunction, and loss of endothelium-dependent vasodilation appears early in the development of this disorder. Nitric oxide (NO) appears to be the principle mediator of erectile function and is generated in part by the sinusoidal endothelium. Vascular endothelial growth factor (VEGF) is an angiogenic growth factor and an endothelial cell-specific mitogen and the actions of VEGF are coupled to NO. In this preliminary study, we investigated whether VEGF could be used to protect endothelial dependent cavernosal relaxation from the atherosclerotic injury induced by a hypercholesterolemic diet.Two groups of New Zealand white adult male rabbits received a 1% cholesterol diet for four weeks, and two groups consumed normal rabbit chow. Half of the rabbits consuming the 1% cholesterol diet received weekly penile injections of 0.3 mg VEGF (n=8), and half injections of normal saline (n=8). Rabbits fed normal chow followed a similar protocol, half received weekly penile injections of 0.3 mg VEGF (n=6) and half were given weekly penile injections of normal saline (n=6). Isometric tension studies (with norepinephrine, acetylcholine, sodium nitroprusside and histamine) were performed on isolated strips of corpora cavernosa. The degree of corporal smooth muscle relaxation in response to ACH and SNP administration was recorded and compared.Significant elevation in serum total cholesterol levels occurred in rabbits receiving 4 weeks of the 1% cholesterol diet (727+/-75.6 mg/dl vs 38.7+/-5.53 mg/dl) P<0.01. There were no significant differences in cavernosal contraction in any group, while cavernosal smooth muscle from rabbits on normal chow retained the ability to relax in response to ACH and SNP in tissue bath. The hypercholesterolemic rabbits receiving VEGF had a significantly higher maximal per-cent relaxation to ACH (111+/-28.9) compared to the hypercholesterolemic rabbits that received NS (77+/-23.1, P<0.001). This difference in percent maximal relaxation to SNP was also present for hypercholesterolemic/VEGF rabbits (129.4+/-24) versus the hypercholesterolemic/NS rabbits (115.0+/-18, P=0.033).In conclusion, intracavernosal injections of VEGF appear to protect corporal endothelium from hypercholesterolemia induced injury, thus preserving endothelial dependent corporal smooth muscle relaxation in hypercholesterolemic rabbit.     

腹腔注射舒芬太尼对SD大鼠气道反应性的影响

目的观察腹腔注射舒芬太尼对SD大鼠气道反应性的影响。方法将32只雄性SD大鼠按随机数字表法分为4组,正常对照为A组(n=8,腹腔注射生理盐水,3 ml);实验组,B1组(n=8,腹腔注射舒芬太尼60μg/kg,3 ml)、B2组(n=8,腹腔注射舒芬太尼90μg/kg,3 ml)和B3组(n=8,腹腔注射舒芬太尼120μg/kg,3 ml)。第1天对4组大鼠进行气道阻力和气道反应性应用非侵入性气道阻力仪测定并作为参考基线。第15天按照分组要求对4组大鼠进行腹腔注射干预措施,测定腹腔注射后1、2、3、5、10 min气道阻力(specific airway resistances,Raw)(cmH2O·s),第30天按照分组要求对4组大鼠进行腹腔注射干预措施,3 min后进行气道反应性测定。采用t检验和Fisher确切概率法对实验数据进行分析。结果实验第1天,与对照组A组比较,B1组、B2组、B3组大鼠腹腔注射前基线气道阻力(3.27±0.74比3.35±0.65、3.30±0.81比3.35±0.65、3.41±0.68比3.35±0.65)差异无统计学意义(t=0.230,P>0.05、t=0.136,P>0.05、t=0.180,P>0.05);实验第15天,与对照组A组干预后1、2、3、5、10 min的气道阻力比较,B1组在1、2 min气道阻力(4.15±1.27比3.95±0.89、4.74±1.05比3.78±0.85)差异无统计学意义(t=0.365,P>0.05、t=2.010,P>0.05),在3、5 min气道阻力(4.92±1.33比3.55±0.69、4.89±1.43比3.47±0.71)差异有统计学意义(t=2.586,P<0.05、t=2.516,P<0.05),在10 min气道阻力(3.86±1.28比3.41±0.69)差异无统计学意义(t=0.875,P>0.05);B2组在1 min气道阻力(4.77±1.15比3.95±0.89)差异无统计学意义(t=1.595,P>0.05),在2、3、5 min气道阻力(6.01±1.47比3.78±0.85、6.95±1.68比3.55±0.69、5.75±1.52比3.47±0.71)差异有统计学意义(t=3.715,P<0.05、t=5.295,P<0.05、t=3.844,P<0.05),在10 min气道阻力(3.95±1.18比3.41±0.69)差异无统计学意义(t=1.117,P>0.05);B3组在1、2、3、5 min气道阻力(5.14±1.25比3.95±0.89、7.91±1.35比3.78±0.85、8.15±1.48比3.55±0.69、7.69±1.23比3.47±0.71)差异有统计学意义(t=2.194,P<0.05、t=7.322,P<0.05、t=7.968,P<0.05、t=8.404,P<0.05),在10 min气道阻力(4.16±1.08比3.41±0.69)差异无统计学意义(t=1.655,P>0.05);实验第30天,4组大鼠腹腔注射后气道反应性阳性率为,与对照组A组(0%)比较,B1组37.5%差异无统计学意义(P>0.05)、B2组75%差异有统计学意义(P<0.05)和B3组87.5%差异有统计学意义(P<0.05)。结论腹腔注射舒芬太尼对大鼠呼吸系统的影响表现在气道阻力增加和气道反应性增高,并呈剂量依赖性。但这种变化在短时间内逐渐增加并达到高峰后,进而逐渐减弱。     

罗哌卡因复合不同剂量舒芬太尼硬膜外分娩镇痛对胎心率的影响

目的研究罗哌卡因复合舒芬太尼硬膜外分娩镇痛对胎心率(FHR)的影响。方法足月产妇132例,随机均分为四组,硬膜外负荷量分别为0.1%罗哌卡因10ml(S0组),0.1%罗哌卡因10ml复合舒芬太尼5μg(S1组)、10μg(S2组)、15μg(S3组);维持剂量均为0.1%罗哌卡因持续背景输注10ml/h,每15分钟追加3ml。记录镇痛前后90min的FHR,FHR加速的数量,早期、变异、迟发FHR减速的数量和持续时间,小振幅FHR变异持续总时间。结果四组产妇分娩过程中硬膜外注药后均未发生心率变化和呼吸抑制情况,对镇痛效果均表示满意。S2组和S3组各有1例产妇在镇痛后出现短暂低血压,输注复方乳酸钠纠正。低血压时S2组出现短于60s的胎心变异减速,S3组出现长于60s的胎心变异减速。四组镇痛后0～30、30～60min时间段单个胎心减速平均值、胎心减速持续总时间较镇痛前30～0min明显延长(P<0.01),且S3组单个胎心减速时间明显长于其它三组(P<0.01)。S3组有2例各出现1次与血压无关且长于60s的变异减速,S3组有3例出现长于60s的变异减速。结论 0.1%罗哌卡因10ml复合5或10μg舒芬太尼作为硬膜外镇痛负荷量对FHR改变是可逆的。     

人参皂苷Rg1对七氟醚麻醉所致幼鼠远期认知功能障碍的保护作用及机制

目的 观察人参皂苷Rg1(ginsenoside Rg1,Rg1)对七氟醚麻醉所致幼鼠远期认知功能障碍的改善作用及其机制.方法 72只新生6日龄C57BL/6J小鼠采用随机数字表法分为4组(每组18只):对照+生理盐水组(Con+NS组)、对照+Rg1组(Con+Rg1组)、七氟醚麻醉+生理盐水组(Sev+NS组)、七氟醚麻醉+Rg1组(Sev+Rg1组).Sev+NS组和Sev+Rg1组分别在出生后6~8 d每天接受3%七氟醚+100%氧气麻醉2 h,Con+NS组及Con+Rg1组小鼠在相应日龄吸入相同时间100%氧气.麻醉前30 min各组分别进行生理盐水(1 ml·kg-1·d-1)或Rg1(10 mg·kg-1·d-1)腹腔注射.各组取12只小鼠于第31~37天行水迷宫实验,行为学测试结束后取海马行Western blot检测突触后密度蛋白95(postsynaptic density 95,PSD-95)含量,其余小鼠(每组6只)于第8天麻醉手术后即刻行ELISA检测海马ATP及活性氧簇(reactive oxygen species,ROS)水平.结果 与Con+NS组比较,Sev+NS组第35~37天水迷宫实验逃避潜伏期[(35.6±4.5)、(28.3±3.5)、(21.9±2.4)s比(45.7±8.1)、(41.9±8.8)、(35.1±12.4)s]明显延长(P<0.05),平台次数[4(8,2)次比2(6,0)次]明显减少(P<0.05),PSD-95水平[(100±6)%比(77±6)%]明显降低(P<0.05),ROS水平[(100±4)%比(121±11)%]明显升高(P<0.05),ATP水平[(100±6)%比(82±7)%]明显降低(P<0.05).Sev+Rg1组与Con+Rg1组比较,31~37 d水迷宫实验逃避潜伏期、 穿越平台次数、PSD-95水平、ROS水平及ATP水平差异均无统计学意义(P>0.05).结论 Rg1可改善七氟醚麻醉所致的幼鼠远期认知功能障碍,其机制可能与抑制氧化应激保护线粒体功能并增强突触可塑性有关.     

真核始动因子4E反义寡核苷酸对人膀胱癌BIU-87细胞中eIF4E和肝素酶表达的影响

目的 探讨真核始动因子4E(eIF4E)反义寡核苷酸(ASODN)对人膀胱癌BIU-87细胞株中eIF4E及肝素酶(HPA)蛋白、mRNA表达的影响. 方法 采用脂质体介导方法分别将2.5、5.0及7.5 μg/ml eIF4E ASODN转染膀胱癌BIU-87细胞,并设立细胞对照组(不转染)、空白对照组(转染空脂质体)及无关对照组(转染无关对照ASODN),分别转染24、48及72 h,采用原位杂交、免疫细胞化学方法分别检测各组BIU-87细胞中eIF4E及HPA蛋白、mRNA表达. 结果 eIF4E ASODN各转染组细胞中eIF4E蛋白及mRNA表达量均较对照组降低(蛋白:2.5 μg/ml组分别为3.55±0.52、3.52 ±0.51、3.22 ±0.44;5.0 μg/ml组分别为3.43±0.47、3.41±0.46、3.19±0.41;7.5 μg/ml组分别为2.36±0.39、2.33±0.37、2.05±0.30.mRNA:2.5 μg/ml组分别为3.19±0.41、3.13±0.39、2.90±0.38;5.0μg/ml组分别为3.07±0.39、2.94 ±0.38、2.27±0.37;7.5 μg/ml组分别为2.22±0.37、2.06±0.30、1.95 ±0.29),与细胞对照组、空白对照组和无关对照组相应时间段、相应浓度组两两比较差异均有统计学意义(P＜0.05),且具有时间和浓度依赖性.eIF4E ASODN各转染组细胞中HPA蛋白及mRNA表达量均较对照组降低(蛋白:2.5 μg/ml组分别为4.44±0.55、4.40±0.54、3.99±0.52;5.0μ g/ml组分别为4.41 ±0.55、4.21±0.53、3.77±0.50;7.5 μg/ml组分别为4.02±0.52、3.98±0.52、2.32±0.37.mRNA:2.5μg/ml组分别为4.12±0.51、3.75±0.50、3.63±0.45;5.0 μg/ml组分别为4.00 ±0.51、3.71±0.50、3.54 ±0.44;7.5 μg/ml组分别为3.87±0.52、3.61 ±0.49、2.08 ±0.30),与细胞对照组、空白对照组和无关对照组两两比较差异均有统计学意义(P＜0.05),且具有时间和浓度依赖性.eIF4E ASODN对HPA蛋白、mRNA表达的抑制效应和对eIF4E蛋白、mRNA表达的抑制效应呈正相关关系(HPA蛋白r=9.23,mRNA r=9.59,P值均＜0.01). 结论eIF4E ASODN可下调膀胱癌BIU-87细胞中eIF4E、HPA蛋白及mRNA的表达,抑制肿瘤细胞的生长、浸润及转移.     

Prostaglandin E1 activates the large-conductance KCa channel in human corporal smooth muscle cells.

The large conductance calcium-sensitive potassium channel (KCa or maxi-K) is an important modulator of human corporal smooth muscle tone, and therefore, erectile capacity. The goal of this investigation was to evaluate the actions of prostaglandin E1 (PGE1), the most widely used and effective drug for the treatment of impotence, on the activity of the KCa channel, a prominent K+ current present in human corporal smooth muscle. Whole-cell patch clamp studies conducted on short-term cultured and enzymatically dissociated human corporal smooth muscle cells, revealed mean resting potentials of -50.8 +/- 2.1 mV (n = 8) and -34 +/- 4 mV (n = 8), respectively. In the attached-patch configuration, the corresponding single-channel slope conductance values for the KCa channel subtype were 173 +/- 4 pS (n = 8) in cultured cells, and 190 +/- 13 pS (n = 3) in freshly isolated myocytes. Furthermore, voltage clamp experiments revealed that relative to control values, the application of PGE1 to cultured cells (3.3 or 33 microM) elicited an apparent increase in both the open probability (Po; ranging from 1.2-23 fold), and the mean open time (5-6 fold) of the KCa channel at membrane potentials of +90 mV and +110 mV. PGE1-induced alterations in KCa channel activity were also observed in freshly isolated corporal myocytes. In the whole cell-recording mode, statistically significant, Charybdotoxin-sensitive (100 nM) 2-3 fold increases in the outward K+ currents were observed in both cultured and freshly isolated corporal myocytes. The presence of a PKA inhibitor (fragment 6-22 amide; 10 microM) in the pipette tip was also associated with a nearly complete ablation of the observed PGE1-induced whole cell K+ currents. Taken together, these data confirm and extend our previous observations, and indicate that PGE1-induced relaxation of human corporal smooth muscle is related, at least in part, to activation of the KCa channel subtype resulting in cellular hyperpolarization.     

Two in one: patient-controlled epidural analgesia (PCEA) to prevent erection and control pain in adult hypospadias-surgery patients.

Following penile surgery, erections are painful and may prejudice the result, because the sutures may not withstand a rigid erection. Therefore, prevention of erection and management of pain are extremely important following hypospadias repair, especially in adult patients. In this prospective study, we aimed to achieve these goals by using an epidural block with patient-controlled analgesia. We allocated 20 adult patients scheduled for hypospadias repair randomly either to receive or not to receive epidural analgesia. Postoperative pain was scored according to a standardised scoring system, based on a 10 point visual analogue scale. In group I (n = 10), analgesia was provided by a 3 ml h(-1) continuous infusion of fentanyl (2 microg) and bupivacaine solution (0.125%) in 1 ml saline via an epidural catheter for the first 3 days. Patient-controlled epidural analgesia was administered with an additional 5 ml of the same solution when the pain score was high (> 4). After 3 days, fentanyl was excluded from the treatment protocol, and analgesia was maintained with bupivacaine (0.125%). In group II (n = 10, control group), an epidural catheter was not inserted, and analgesia was maintained with pethidine (1 mg kg(-1)). Pain management was found to be more effective in group I. No erections occurred in group I, but the erection rate in group II was mean +/- s.d. = 1.7 +/- 0.2. The differences were found to be statistically significant (P<0.05). We highly recommend the technique described here, which offers efficient analgesia and control of erection in adult hypospadias patients.