MMP-9、TIMP-1在曲张大隐静脉中的表达及意义

目的：研究基质金属蛋白酶9（MMP-9）与基质金属蛋白酶组织抑制剂1（TIMP-1）在曲张大隐静脉中的表达及其与发病机制的关系。方法：收集佳木斯大学附属第一医院2012-11～2013-09,血管外科手术治疗的大隐静脉曲张患者31例（40条肢体）,其中男19例,女12例,年龄35～71岁,平均51.25岁。标本均取自大隐静脉主干曲张最显著部位。根据CEAP临床分级法,将40条患肢分为C1～C3组（24条肢体）和C4～C6组（16条肢体）。对照组选择同期佳木斯大学附属第一医院血管外科和骨外科,因外伤或动脉硬化闭塞症（排除下肢静脉曲张和静脉曲张家族史）而截肢的正常大隐静脉10例。其中男7例,女3例,年龄25～67岁,平均48.10岁。实验组与对照组均采用免疫组织化学染色法检测MMP-9、TIMP-1蛋白在大隐静脉中的表达。结果：MMP-9可在正常大隐静脉壁中少量表达,而在曲张大隐静脉壁中表达明显增高（P〈0.05）,C4～C6组和C1～C3组表达率无统计学意义（P〉0.05）。TIMP-1也可在正常大隐静脉壁中表达,而在曲张大隐静脉壁中表达明显增高（P〈0.05）,C4～C6组和C1～C3组表达率无统计学意义（P〉0.05）。结论：MMP-9、TIMP-1的异常表达可能参与了大隐静脉曲张的血管重塑过程。     

Deletion of the vaccinia virus F13L gene results in a highly attenuated virus that mounts a protective immune response against subsequent vaccinia virus challenge

Vaccinia virus F13L encodes the envelope protein p37, which is the target of the anti-pox virus drug ST-246 (Yang et al., 2005) and that is required for production of extracellular vaccinia virus. The F13L (p37)-deleted (and ST-246 resistant) vaccinia virus recombinant (Vac-ΔF13L) produced smaller plaques than the wild-type vaccinia (Western Reserve vaccinia). In addition, Vac-ΔF13L proved, when inoculated either intravenously or intracutaneously in both immunocompetent and immunodeficient (athymic nude or SCID) mice, to be severely attenuated. Intravenous or intracutaneous inoculation of immunocompetent mice with the ΔF13L virus efficiently protected against a subsequent intravenous, intracutaneous or intranasal challenge with vaccinia WR (Western Reserve). This was corroborated by the observation that Vac-ΔF13L induced a humoral immune response against vaccinia following either intravenous or intracutaneous challenge. In conclusion, F13L-deleted vaccinia virus may have the potential to be developed as a smallpox vaccine.     

兔主动脉球囊损伤后TIMP～1的mRNA动态表达

目的 探讨基质金属蛋白酶组织抑制因子-1(TIMP-1)mRNA在球囊损伤后的改变及可能作用。方法 在兔腹主动脉球囊损伤的模型上，应用RT—PCR的方法，观察TIMP-1 mRNA在球囊损伤后不同时间的表达情况。结果 TIMP-1 mRNA球囊损伤后第1天表达开始逐渐升高，第4天达高峰。结论 TIMP-1 mRNA球囊损伤后表达逐渐升高，第4天达高峰可能与再狭窄中、后期新生内膜形成及血管重塑有关。     

Inhibition of HSV-1 and vaccinia virus replication by cephalosporin derivatives

Derivatives of beta-lactam antibiotics of the cephalosporin type at 0.02-1 mM concentrations interfered with in vitro replication of two DNA-containing viruses, herpes simplex I and vaccinia, but showed no effects on two RNA-viruses, lymphocytic choriomeningitis virus and vesicular stomatitis virus, or on cell viability. The exact structure of the active compounds remains unknown, but opening of the beta-lactam ring appears to be a prerequisite for their formation. Whereas cephalosporin derivatives were most active, no active products were obtained from penicillins and monobactams. The potential of these unexpected antiviral effects of widely used beta-lactam antibiotics remains subject of further study.     

Effects of cidofovir treatment on cytokine induction in murine models of cowpox and vaccinia virus infection

Cytokine profiles during cowpox and vaccinia (WR strain) virus infections were characterized in intranasal (i.n.) and intraperitoneal (i.p.) models in BALB/c mice. The time-course of induction and effects of cidofovir treatment on interferon (IFN)-gamma, IFN-gamma inducible protein (IP)-10, interleukin (IL)-6, and monocyte chemoattractant protein (MCP)-1 were determined. The four mouse infection models have distinct patterns of cytokine induction. Cowpox virus i.p. and vaccinia virus i.n. infections showed increased induction throughout the time studied. Cowpox virus i.n. infection resulted in delayed induction of IFN-gamma and IP-10. Cytokine levels were fairly constant during vaccinia virus i.p. infections. Cidofovir treatment (100mg/kg/day i.p. for 2 days) significantly suppressed certain cytokine (IFN- gamma, IL-6, IL-10, IL-11, IP-10, LIF, MCP-1, MCP-3, MCP-5, MIP-1 gamma, and TIMP-1) levels to near normal relative to uninfected animals, as well as prevented mortality and reduced virus titers significantly. Characterization of cytokine responses has implications for understanding the immune responses and pathogeneses of viral infections in these mouse models.     

Effects of cidofovir on the pathogenesis of a lethal vaccinia virus respiratory infection in mice

Intranasal infection of BALB/c mice with the WR strain of vaccinia virus leads to pneumonia, profound weight loss, and death. Although the major sites of virus replication are in the lungs and nasal tissue, dissemination of the virus to other visceral organs and brain occurs via the blood. In this report the effects of cidofovir on the pathogenesis of the infection was studied. Mice were infected intranasally with virus followed 1 day later by a single intraperitoneal treatment with cidofovir (100 mg/kg) or placebo. Placebo-treated mice were dead by day 8, whereas all cidofovir-treated animals survived through 21 days. Cidofovir treatment did not prevent profound weight loss from occurring during the acute phase of the infection, but the mice gained weight quickly after the 8th day. Significantly higher arterial oxygen saturation levels, as determined by pulse oximetry, were seen in cidofovir-treated animals compared to placebos on days 4-7. Cidofovir treatment markedly improved lung consolidation scores and prevented lung weights from increasing during the infection. Virus titers in lungs and nasal tissue were high starting from the first day of the infection, whereas the titers in liver, spleen, brain, and blood was low for 3 days then markedly rose between days 4 and 6. Lung and nasal virus titers were reduced 10-30-fold by cidofovir treatment on days 2, 4 and 6. Virus titers in the other tissues and blood at their peak (day 6) were 30- to >1000-fold less than in tissues of placebos. These results illustrate the ability of a single cidofovir treatment to control the pathogenesis of an acute lethal infection in various tissues during the vaccinia virus infection in mice.     

The antiviral activity exerted by vaccinia virus on the growth of herpes simplex virus in BS-C-1 cells

The growth of herpes simplex virus type 2 (HSV-2) in BS-C-1 cells, was inhibited following super-infection with vaccinia virus. This inhibition was efficiently induced by both the intracellular mature virus (IMV) form of vaccinia virus and the extracellular enveloped virus (EEV), containing an additional external viral membrane. Treatment of vaccinia IMV with the detergents NP-40, Brij-58 or n-octyl-alpha-D-glucopyranoside, abolished its ability to inhibit the growth of HSV-2. Ultraviolet irradiation of vaccinia virus, that completely inactivated the infectivity of the virus, resulted in partial loss of the capability to inhibit the growth of HSV-2: 16-fold more irradiated virus was needed for the inhibition. Electron microscopy showed that the irradiated vaccinia virus adsorbed and penetrated into the HSV-infected cells but remained morphologically intact within the cells for at least 22 h. When the steps in the growth of HSV affected by the irradiated vaccinia virus were followed, it was found that while the synthesis of HSV DNA was partially decreased, the synthesis of HSV proteins was very strongly inhibited and virus particles were not formed.     

Surgical trauma and vein graft failure: further evidence for a role of ET-1 in graft occlusion

The saphenous vein (SV) is the most commonly used conduit for coronary artery bypass surgery. However, using traditional techniques, the occlusion rate for the SV is high, with over 50% of grafts failing within 10 years. In conventional coronary artery bypass surgery the SV is exposed to considerable damage during preparation for grafting. Recently, an increased graft patency has been described using a 'no-touch' technique, whereby the vein is prepared with minimal vascular trauma. There is evidence that the success of this form of coronary artery bypass surgery is a result, at least in part, of the retention of tissue-derived nitric oxide. We have examined the effects of conventional SV harvesting on vessel morphology, cell proliferation, endothelin-1 and its receptors. Considerable damage was observed in veins prepared using conventional surgery compared to 'no-touch' veins. The vessel wall exhibited evidence of surgical trauma, with regions of denudation of the luminal endothelium caused by distension. Endothelin-1 and endothelin-A receptors were present at subintimal regions of conventional SV segments where proliferating cells were identified. Endothelial endothelin-B receptors were also revealed that were absent at areas of distension-induced damage to the endothelium. These results suggest that endothelin-1 plays a role in vein graft failure, predominantly via the endothelin-A receptor.     

Effects of atorvastatin therapy on protein oxidation and oxidative DNA damage in hypercholesterolemic rabbits

ObjectiveOur aim was to clarify the effects of hypercholesterolemic diet and administeration of atorvastatin on lipid peroxidation, protein oxidation and oxidative DNA damage in male New Zealand white rabbits.MethodsWe determined malondialdehyde (MDA), protein carbonyl (PCO) and total thiol (T-SH) levels in plasma and liver tissue, glutathione (GSH) levels in erythrocyte and liver tissue, and 8-hydroxy-2-deoxyguanosine (8-OHdG) levels in plasma. Twenty rabbits were randomly divided into two groups and fed with a high-cholesterol diet (fortified with 1% cholesterol) for 4 weeks. Such rabbits were subjected to either (Group 1) a high-cholesterol diet non-supplemented with atorvastatin (n = 10) or (Group 2) a high-cholesterol diet supplemented with atorvastatin (0.3 mg atorvastatin per day/kg body weight) for 4 weeks (n = 10). A control group (n = 5) (Group 3) was fed a cholesterol free diet for 4 weeks. Colorimetric methods were used to determine the level of the oxidative stress markers, except 8-OHdG, which was measured by ELISA.ResultsRabbits were fed with the high-cholesterol diet alone (Group 1) showed higher levels of lipid profile and oxidative protein and DNA damage than compared with dose of the control group (Group 3). Atorvastatin therapy has substantially beneficial effects on oxidative protein and DNA damage in hypercholesterolemic rabbits.ConclusionsThe current findings will, we hope, lead to a new insight into the pathogenesis of atherosclerosis. On the other hand inhibition of protein oxidation and DNA oxidation in the plasma by atorvastatin may be one of the pleiotropic effects of statins, and thus the underlying mechanism needs to be further clarifications.     

Effects of four antiviral substances on lethal vaccinia virus (IHD strain) respiratory infections in mice

Intranasal infection of BALB/c mice with the IHD strain of vaccinia virus was found to cause pneumonia, profound weight loss and death. Cidofovir, hexadecyloxypropyl-cidofovir (HDP-CDV), the diacetate ester prodrug of 2-amino-7-[(1,3-dihydroxy-2-propoxy)methyl]purine (HOE961), and ribavirin were used to treat the infections starting 24h after virus exposure. Single intraperitoneal (i.p.) cidofovir treatments of 100 and 30 mg/kg led to 90-100% survival compared with no survivors in the placebo group, whereas a 10 mg/kg dose was ineffective. The 100 mg/kg treatment reduced lung and snout virus titres on day 3 of the infection by 20- and 8-fold, respectively. Mean arterial oxygen saturation levels in these two cidofovir treatment groups were significantly higher than placebo on days 4 through 6 of the infection, indicating an improvement in lung function. Effects of cidofovir on viral pathogenesis were studied on days 1, 3 and 5 of the infection, and demonstrated statistically significant reductions in lung consolidation scores, lung weights, lung virus titre and snout virus titres on days 3 and 5. Cidofovir treatment also reduced virus titres in other tissues and body fluid, including blood, brain, heart, liver, salivary gland and spleen. HDP-CDV was given by oral gavage at 100, 50 and 25mg/kg doses one time only, resulting in 80-100% survival. Lower daily oral doses of 10 and 5mg/kg per day given for 5 days protected only 30% of animals from death. Oral doses (100, 50 and 25 mg/kg per day) of HOE961 for 5 days protected all animals, whereas equivalent oral doses of ribavirin were completely ineffective. The rapidity of recovery from weight loss during the infection was a function of dose of compound administered. These data indicate the utility of parenteral cidofovir, oral HDP-CDV and oral HOE961 in treating severe respiratory infections caused by this virus.     

云芝多糖B对大鼠单核细胞趋化蛋白—1基因表达的影响

目的:探究野生云芝多糖水溶性新组分CVPS-B对大鼠脾细胞单核细胞趋化蛋白-1(MCP-1)基因表达的影响.方法:以β-actin为内标准物,用逆转录聚合酶链式反应(RT-PCR)检测CVPS-B分别对正常情况下以及脂多糖(LPS)诱导下大鼠脾细胞MCP-I基因表达的影响,并对RT-PCR产物进行测序,以证实其特异性.结果:(1)正常情况下大鼠脾细胞MCP-1 mRNA的表达(MCP-1/β-actin的比值)生理盐水对照组为1.4±0.3;CVPS-B三个剂量组(10、30和50mg·kg~(-1)·d~(-1),ip,连续4d)分别为:1.6±0.4、1.7±0.5和1.5±0.4,后三组与对照组无显著差异(P>0.05);(2)大鼠腹腔给药LPS(10μg·kg~(-1)可使脾细胞MCP-1 mRNA的表达增加114％.(3)CVPS-B4个剂量组(5、10、30和50mg·kg~(-1)·d~(-1),ip,连续4d)可使LPS(10μg·kg~(-1),ip)诱导的脾细胞MCP-1 mRNA的表达分别减低51％,70％,84％和99％(n=6).结论:CVPS-B可预防性抑制LPS对大鼠脾细胞MCP-1基因表达的诱导作用,且呈剂量依赖性,但对正常情况下大鼠脾细胞MCP-1 mRNA的表达则无明显影响.     

Endothelin-1 (ET-1) and vein graft failure and the therapeutic potential of ET-1 receptor antagonists

Despite the exploration of a large number of disparate drugs in animal models and clinical trials, no pharmacological intervention, with the exception of aggressive lipid lowering therapy has reduced late vein graft failure in man. The importance of devising more effective strategies is exemplified by the enormous economic consequences of vein graft failure. Worldwide, there are currently more than 1,000,000 coronary artery bypass graft surgery (CABG) operations a year, the same number of patients undergoing infrainguinal bypass for vascular diseases of the lower limb. The pathophysiology of vein graft failure is complex, involving disparate factors that include adhesion of platelets and leukocytes, rheological forces, metalloproteinase expression, proliferation and migration of vascular smooth muscle cells, neointima formation, oxidative stress, hypoxia and neural re-organisation. Although this diverse etiology may seem to preclude any single drug type as being effective in mediating vein graft failure: one factor that is involved in every facet of vein graft pathobiology is endothelin-1 (ET-1). As such a single drug type (ET_A antagonist) may prove to be the magic bullet in this scenario. Thus, in this review, we will consider the etiology of vein graft disease in relation to ET-1 and will then present an argument (with evidence) that specific ET_A receptor antagonists constitute a potentially effective means of preventing vein graft failure.     

基质金属蛋白酶组织抑制因子-1、2在宫颈鳞癌中的表达及意义

目的:探讨基质金属蛋白酶组织抑制因子(TIMP)-1和TIMP-2在宫颈鳞癌组织中的表达及其与宫颈癌临床病理特征的关系。方法:采用免疫组织化学MaxvisionTM法,检测46例宫颈鳞癌,34例宫颈原位癌(CINⅢ)和20例正常宫颈上皮(NCE)组织中TIMP-1、TIMP-2的表达,分析其表达与宫颈鳞癌的临床分期、病理类型及有无淋巴结转移的关系。结果:宫颈鳞癌、原位癌和正常宫颈上皮中TIMP-1的阳性表达率分别为54.3%(25/46)、73.5%(25/34)和20.0%(4/20)。TIMP-2为60.9%(28/46)、50.0%(17/34)和20.0%(4/20)。TIMP-1和TIMP-2蛋白阳性表达率在鳞癌组及原位癌组明显高于对照组,差异有统计学意义(P<0.01);在不同的临床分期和病理类型宫颈鳞癌患者中,TIMP-1和TIMP-2的阳性表达率差异无统计学意义(P>0.05);TIMP-2在宫颈鳞癌无淋巴结转移患者的阳性表达率较有转移患者高(P<0.05),而TIMP-1未发现此现象。结论:TIMP-1和TIMP-2在子宫颈鳞癌浸润和转移中起重要的作用。     

辛伐他汀对自体移植静脉内膜增生的抑制作用

目的 研究辛伐他汀对自体移植静脉内膜增生的影响.方法 48只健康成年家兔随机均分为辛伐他汀治疗组(A组)和生理盐水对照组(C组).建立自体静脉旁路移植模型,术后3、7、28 d切取移植静脉,观察组织病理变化,测量新生内膜厚度及面积,检测静脉壁增殖细胞核抗原(PCNA)表达,RT-PCR检测静脉壁基质金属蛋白酶(MMP)2、MMP-9 mRNA表达,明胶酶谱法检测MMP-2、MMP-9活性.结果 与C组比较,A组术后内膜增生、移植静脉壁阳性细胞数以及MMP-2、MMP-9 mRNA表达和活性均明显减少(P<0.05).结论 辛伐他汀可抑制自体移植静脉内膜增生,其作用机制可能是通过抑制血管平滑肌细胞(VSMC)增殖并限制其向内膜迁移而实现的.