Effect of Icariin on Cyclic GMP Levels and on the mRNA Expression of cGMP-binding cGMP-specific Phosphodiesterase (PDE5) in Penile Cavernosum

Penile erection, which is characterized by the relaxation of arterial and cavernous smooth muscles, is mediated by nitric oxide (NO). After its release from endothelial cells and nerve endings within the penis, NO diffuses into the smooth muscle cells and activates the soluble guanyl cyclase, which then catalyzes the conversion of GTP to cGMP[1, 2]. Elevated cGMP concentrations activate pro- tein kinase G (PKG), which in turn phosphorylates gap junctions, potassium (K ) channels, and ca…     

Effect of Berberine on the mRNA Expression of Nitric Oxide Synthase （NOS） in Rat Corpus Cavernosum

Penile erection is a complex neurovascularprocess that involves relaxation of the corpus cav ernosum smooth muscle[1]. Relaxation of cavern osal smooth muscle leads to engorgement of bloodin the corpus cavernosum that results in an in crease in intracavernosal pressure and thus in tu mescence. Many studies have shown that nitric ox ide (NO) released from cavernous nerves and endo thelium is a key factor for penile erection. TheNO cGMP signaling pathway is…     

Effect of Antisense FosB and CREB on the Expression of Prodynorphin Gene in Rats with Levodopa-induced Dyskinesias

The effects of antisense FosB and CREB intra-striatum injection on the expression of prodynorphin (PDyn) gene in striatal neurons of Levodopa-induced dyskinesias (LID) rats with Parkinson disease (PD) were explored. PD model in rats was established by 6-OHDA microinjection stereotaxically. The rats were treated with chronic intermittent Levodopa celiac injection for 28 days to get the LED rats. Antisense FosB and cAMP response element-binding protein (CREB) were injected into striatum of all rats respectively. In situ hybridization was used to measure the changes in the expression of PDyn mRNA in striatum and behavior changes were observed. The results showed after administration of antisense FosB, abnormal involuntary movement (AIM) was decreased and the expression of PDyn mRNA in striatum was increased in LID rats as compared with sense FosB group (P<0.01, respectively). As compared with the control group, the expression of PDyn mRNA in striatum was decreased by antisense CREB-treated LID group (P<0.01) and compared with sense CREB treated LID group, antisense CREB-treated LID group showed no changes in AIM scores and the expressions of PDyn mRNA (both P>0.05). In conclusion, FosB protein, which replaced the CREG, could regulate the expression of PDyn mRNA and play critical role in the pathogenesis of LID.     

Expression and characterization of Gag protein of HIV-1 （CN） in Pichia pastoris

To express the core protein of HIV-1 of Chinese prevalent strata （HIV-1[CN]） in Pichia pastoris, the fulllength gag gene was inserted into the secretory expression vector pHILS1. Linearized recombinant plasmid pHILGAG by Sal I was electrotransformed into the yeast strain GS115, and the yeast transformants were identified by PCR. To induce the interest protein to be expressed, the PCR positive transformants were inoculated in the medium of BMGY and BMMY, mRNA of the strain was detected by RT-PCR, and the expressed protein was analyzed by SDS-PAGE, Western blotting and thin layer scanning. mRNA （1.3 kb） was amplified by RT-PCR. SDS-PAGE and Western blotting analysis showed that the molecular mass of the expressed protein was 55 kDa, which was similar to the expected value, and the expressed protein could react with McAb to HIV-1 p24. Thin layer scanning analysis demonstrated that the whole amount of the expressed protein was approximately 13% of the soluble protein in the supernatant. The recombinant yeast had good genetic stability. The optimal expression conditions of the engineering yeast were as follows： BMMY medium, 80%-90% of dissolved oxygen, 1% methanol, and 3-day-cultivation course. Gag proteins were expressed under the optimal expression condition and purified via gel filtration chromatography. The purity of the interest protein was up to 85%. After the purified proteins were inoculated into BALB/c mice, the anti-HIV-1 antibodies in the immunized mice could be detected by Western blotting.     

Influence of L-arginine on the Expression of eNOS and COX2 in Experimental Pulmonary Thromboembolism

The influence of L-arginine on endothelial nitric oxide synthase (eNOS) and cyclooxy-genase 2 (COX2) was observed in experimental pulmonary thromboembolism and the action mechanism on pulmonary thromboembolism was explored. Wistar rats were randomly divided into control group, model group and treatment group. Pulmonary thromboembolism models were established by auto-blood back transfusion, and L-Arg 100 mg/kg was intraperitoneally injected after successful model preparation. The animals were sacrificed at 3 h, 1 day, 3 days and 7 days after embolism. Plasma NO, TXB2 and 6-Keto-PGF1αwere detected. The expression of eNOS and COX2 protein and mRNA in pulmonary tissues was detected by immunohistochemistry and RT-PCR respectively. The results showed that pulmonary thrombosis could be seen post pulmonary embolism and inflammatory reaction was significant. Plasma NO was decreased (P<0.01), and the levels of TXB2, 6-Keto-PGF1αand T/P ratio were all elevated. The expression of eNOS protein and mRNA in the pulmonary tissue was down-regulated (P<0.05), while that of COX2 protein and mRNA was upregu-lated (P<0.01). In treatment group, the level of NO was increased, the levels of TXB2 and T/P ratio were decreased, but the level of 6-Keto-PGF1αwas increased. The expression of eNOS protein and mRNA in pulmonary tissue was upregulated (P<0.05), while that of COX2 protein and mRNA was down-regulated (P<0.05). In conclusion, L-arginine can educe the role of pulmonary tissue protection through up-regulating the expression of intra-pulmonary NOS and down -regulating COX2 in pulmonary thromboembolism.     

Relationship between the Apoptosis and Notch-1 Expression in Acute Pancreatitis

In order to investigate the expression of Notch-1 in rats with acute pancreatitis (AP) and its relation with apoptosis of pancreatic cells, mild AP (MAP) and severe AP (SAP) models were es-tablished by retrograde injection of different concentrations of sodium taurocholae into pancreatic duct. The apoptosis index and the expression of Notch-1 protein and mRNA were detected by using TUNEL, Western blot and real-time PCR in MAP and SAP at different time intervals. The results showed that in MAP group the apoptosis index was significantly elevated during 4 to 24 h after in-duction of pancreatitis, but there was no significant difference among different time intervals. In SAP group, the apoptosis index reached the peak at 4th h after induction of pancreatitis, then gradually de-clined. There was significant difference in apoptosis index between MAP and SAP groups. Starting from 4th after induction of pancreatitis, the expression of Notch-1 in both MAP and SAP group was increased at different time intervals, but that in SAP group was significantly higher than in MAP group (P<0.05). The expression of Notch in MAP and SAP groups reached the peak at 12th and 8th h respectively after induction of pancreatitis. It was concluded that there was significant difference in apoptosis index and the Notch-1 expression in different types of AP. The overexpression of Notch-1 could aggravate AP by inhibiting the apoptosis of pancreatic cells.     

Expression of TLR9 and Its mRNA in the Lesions of Lichen Planus

To investigate the role of toll-like receptor 9 (TLR9) in the pathogenesis of lichen planus, the expressions of TLR9 and its mRNA in the lesional skin of lichen planus were detected by immu- nohistochemical technique (SP) and RT-PCR. As control, normal skin of healthy volunteers was also tested. The immunohistochemical study showed that the expression of TLR9 in the lesional skin of lichen planus was significantly higher than that in the normal controls. The results of RT-PCR showed that both skin lesions and normal controls had TLR9 expression. In skin lesions, the expression level of TLR9 mRNA was 1.6075±0.0930, which was significantly higher than that in normal controls (P<0.001). These findings indicated that up-regulated expression of TLR9 and its mRNA might be involved in the pathogenesis of lichen planus.     

Changes of Expression of Stretch-activated Potassium Channel TREK-1 mRNA and Protein in Hypertrophic Myocardium

Cardiac hypertrophyis an adaptive response tochronic pressure or volume stress.Pressure over-load-induced concentric hypertrophy is believed tohave a compensatory function through reducingwall stress and oxygen consumption.At the sameti me,ventricular hypertrophy is associated withsignificantlyincreased risk of heart failure and ma-lignant arrhythmia[1].During the progression ofventricular hypertrophy,stretch-activated potassi-umchannel plays ani mportant role in signal trans-duction between m…     

Transfection of Antisense Oligodeoxynucleotide Inhibits Heparanase Gene Expression and Invasive Ability of Human Pancreatic Cancer Cell in vitro

Cancer cell invasion and metastasis are criticalsteps in pancreatic cancer progression that worsenthe prognosis.They are facilitated via disassemblyof the main structural elements of basement mem-branes(BM)and extracellular matrix(ECM)byenzymes produced by the cancer cells[1].Hepara-nase(HPSE)is an endoglycosidase responsible forthe degradation of the heparan sulfate proteoglycan(HSPG),a major component of ECM and BM.HPSE mediates tumor invasion pri marily throughthe direct cleavage of…     

疏肝益阳胶囊对动脉性勃起功能障碍大鼠一氧化氮合酶通路及5型磷酸二酯酶表达的影响

目的 研究中药疏肝益阳胶囊治疗大鼠勃起功能障碍(ED)的分子机理.方法 选取3月龄成年雄性SD大鼠,采用双侧髂内动脉结扎法制造勃起障碍模型.设假手术组,模型组,西地那非组[10.5 mg/(kg·d)],疏肝益阳小、大剂量治疗组[0.5、1g/(kg·d)],灌胃给药,疗程30 d.疗程结束后取阴茎海绵体组织,荧光定量...     

Influence of erectile dysfunction course on its progress and efficacy of treatment with phosphodiesterase type 5 inhibitors

Background Erectile dysfunction （ED） is a common impairment among older men, and the prevalence rates increase sharply after age of 60 years. Most studies have focused on the prevalence rate or dangerouse factors. The aim of this study was to investigate the basic epidemiologic data about ED patients with different ED courses. The purpose of this researth was to understand the therapeutic effect of phosphodiesterase type 5 inhibitor （PDE5-1） and see how and why the ED course impact the progress of ED and the therapeutic effect of PDE5-1 treatment. Methods From June 2008 to June 2009, 4252 questionnaires （Quality of Erection Questionnaire, QEQ） were gathered from 46 centers by urology or andrology doctors all around China. Patients with ED （age 〉 20 years） filled in first half of the questionnaires when they came for the first time, and then completed the second half 4 weeks after PDE5-1 therapy. Results ED courses of most patients were less than 5 years （〈5 years, 74.0%; 5-10 years 20.8%; 〉10 years, 5.2%）. As ED course increasing, the incidence of the risk factors of ED, such as smoking, drinking, hypertension, diabetes, heart disease and hyperlipidemia also increase （P 〈0.01）. PDES-I was effective in improving the quality of sexual activities （P 〈0.01）. Administration of PDE5-1 improves satisfaction, enjoyment and frequency of sexual activities. The longer the ED course, the worse the therapeutic effect （〈5 years, 96.1%; 5-10 years, 94.9%; 〉10 years, 89.0%） （P 〈0.01）. Conclusions The ED course greatly affected the therapeutic effect of PDE5-1, the patients with ED should consult doctor at early stage of the disease. Admistration of PDE5-1 effectively improves the penile erection and the quality of sexual life of the patients hence should be considered as first-line medicine in the treatment of ED.     

他达拉非半量隔日口服治疗ED的疗效观察

目的:评估他达拉非半量(10 mg)隔日口服治疗勃起功能障碍(ED)的疗效.方法:选择158例符合美国国立卫生研究院诊断标准的ED患者,随机分为他达拉非(希爱力)组和复方玄驹胶囊组两组,分别接受3个月的治疗,治疗后各组又随访3个月以上.采用勃起功能障碍症状评分(international index of erectile function-5,IIEF-5)评估治疗前后及继续随访3个月后各组的疗效,同时比较患者性生活日记中插入成功率和性交中保持勃起至完成性交的成功率.采用焦虑自评量表评价焦虑状态.结果:治疗前两组IIEF-5评分分别为(9.96±4.21)、(9.69±4.35)分,差异无统计学意义(P＞0.05).治疗3个月后IIEF-5评分分别为(20.38±4.18)、(16.08±3.26)分,随访至6个月时IIEF-5评分分别为(18.16±2.31)、(11.98±3.97)分,差异均有统计学意义(P＜0.01),而且两组间在治疗后和6个月随访时的评分差异也具有统计学意义(P＜0.05).药物治疗后,患者性生活日记中插入成功率和保持勃起的成功率明显提高,希爱力组高于复方玄驹胶囊组.焦虑评分值两组治疗后明显低于治疗前(P＜0.05).结论:他达拉非隔日半量口服可有效治疗ED,且疗效优于传统中药,可作为临床上的一线治疗方法.     

硝普钠阴茎海绵体注射治疗阳萎

目的 探讨ＮＯ供体－硝普钠对促使阴茎勃起的作用。方法 选择 ４２例阳萎患者,采用硝普钠进行阴茎海绵体内注射（ＩＣＩ）,并选择罂粟碱／酚妥拉胆进行对照。结果 硝普钠ＩＣＩ后：１．阴茎长度,周径等明显改变,增加值与对照相比无显著差异,２．Ｖｉｒａｇ硬度计点表明硝普钠与罂粟碱／酚妥拉明效果之间无显著差异。３．所有测试患者无１例出现低血压或局部不适等副反应,与罂粟碱／酚妥拉明相比各有优劣,但总体差异不大。     

一氧化氮合酶在大鼠糖尿病性勃起功能障碍中的作用研究

目的:研究一氧化氮合酶(NOS)在糖尿病性勃起功能障碍中的作用。方法:对正常对照组(n=10)、糖尿病组(DM组,n=10)和链脲佐菌素组(STZ组,n=10)大鼠于造模9周后进行阿朴吗啡阴茎勃起实验,用酶联免疫吸附试验(ELISA)检测阴茎组织中一氧化氮合酶(NOS)活性。结果:DM组大鼠阴茎勃起率较正常对照组及STZ组明显降低(P<0.01),NOS活性亦显著降低(P<0.01),但正常对照组与STZ组之间的差异均无统计学意义(P>0.05)。结论:糖尿病可引起勃起性功能障碍,NOS活性的下降可能参与了DM时勃起性功能障碍的发生。     

糖尿病性大鼠阴茎白膜弹性纤维改变对勃起功能的影响

目的 了解糖尿病大鼠白膜中弹性纤维改变对勃起功能的影响.方法 利用链脲佐菌素(STZ)制作糖尿病及糖尿病性勃起功能障碍模型,40只SPF级SD雄性大鼠,先按时间随机分为两组(每组20只),分别是注射STZ后7周组和4周组,再按处理因素即给予大鼠腹腔注射STZ(60mg/kg)是否成模,分为对照组(未注射STZ,5只)、糖尿病性勃起功能障碍模型组、糖尿病性非勃起功能障碍组、未成模组.利用维多利亚蓝-丽春红染色法对不同处理组的标本进行弹性纤维染色,利用彩色图文系统进行分析,以累积光密度(IOD)作为测量指标.结果 各处理组间I0D存在显著性差异(E=10.433,P＜0.001),糖尿病性勃起功能障碍组IOD均小于对照组、糖尿病非勃起功能障碍组及未成模组(P＜0.05),糖尿病非勃起功能障碍组的IOD小于对照组(P＜0.05),不同时间点间的IOD不存在显著性差异(F=0.685,P=0.415),不存在交互效应(F=0.905,P=0.452).结论 糖尿病可以导致白膜弹性纤维减少;白膜中的弹性纤维在勃起过程中起重要作用;白膜中弹性纤维的减少将导致勃起功能障碍.     

影响男性阻塞性睡眠呼吸暂停综合征患者阴茎勃起功能障碍的因素

目的：探讨引起男性阻塞性睡眠呼吸暂停综合征（OSAS）患者勃起功能障碍（ED）的因素及其可能机制。方法：经多导睡眠监测仪（PSG）及同步夜间阴茎勃起（NPT）测定,90例男性OSAS患者中合并ED患者48例（ED组）,无ED者42例（OSAS组）。所有研究对象于次日清晨测定血清睾酮（TT）、促泌乳素（PRL）、雌二醇（E2）、卵泡刺激素（FSH）、促黄体生成素（LH）、血清一氧化氮（NO）、一氧化氮合酶（NOS）浓度。结果：2组间BMI差异有统计学意义（P＜0.05）,而性激素水平差异均无统计学意义（P＞0.05）,但ED组患者的NO、NOS水平较OSAS组患者明显降低（P＜0.05）,且logistic回归分析发现,肥胖、TT和NO是影响男性OSAS勃起功能障碍的主要因素（P＜0.05）。其中肥胖是危险因素,而TT、NO则是保护因素。结论：男性OSAS患者发生ED可能与肥胖、TT和NO有关,OSAS患者体内NO水平可能在维持其勃起功能中起着关键性作用。     

Role of nitric oxide and inducible nitric oxide synthase in human abdominal aortic aneurysms: a preliminary study

Background Nitric oxide （NO） is an important mediator in the pathophysiology of many vascular diseases. However, the definite role of NO in human abdominal aortic aneurysm （AAA） formation is unclear. The aim of this study was to investigate production of NO and expression of inducible nitric oxide synthase （iNOS）, and their possible role in AAA. Methods A total of 28 patients with AAA, 10 healthy controls, and 8 patients with arterial occlusive disease were enrolled into this study. Standard colorimetric assay was used to examine NO concentration in plasma from patients with AAA and normal controls, and in cultured smooth muscle cells （SMCs）. Expression of iNOS in aortas and cultured SMCs were detected by immunochemistry. The correlation of iNOS expression with age of the patient, size of aneurysm, and degree of inflammation was also investigated by Cochran-Mantel-HaenszelX^2 test and Kendall＇ Tau correlation. Results Expression of iNOS increased significantly in the wall of aneurism in the patients with AAA compared to the healthy controls （P〈0.05） and the patients with occlusive arteries （P〈0.05）. iNOS protein and media NOx （nitrite＋nitrate） also increased in cultured SMCs from human AAA （n=4, P〈0.05）, while plasma NOx decreased in patients with AAA （n=25） compared to the healthy controls （n=20）. There was a positive correlation between iNOS protein and degree of inflammation in aneurismal wall （Kendall coefficient=0.5032, P=0.0029） Conclusions SMCs and inflammatory cells were main cellular sources of increased iNOS in AAA, and NO may play a part in pathogenesis in AAA through inflammation.