宫颈癌组织中的PTEN, VEGF和CD105的表达及临床意义

目的:探讨宫颈癌组织中的人第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)、血管内皮生长因子(VEGF)和内皮因子(CD105)的表达及临床意义。方法:收集2010年1月至2016年2月我院收治的宫颈癌患者组织标本320例和同期正常宫颈组织标本100例,采用SP免疫组化法检测宫颈癌和正常宫颈组织中的PTEN、VEGF和CD105,分析各指标在宫颈癌、正常宫颈组织及不同类型宫颈癌组织中的表达,并采用Sperman分析三者相关关系。结果:宫颈癌组织VEGF阳性率、CD105-MVD高于正常宫颈组织,而PTEN阳性率低于正常宫颈组织,差异均有统计学意义(P0.05);宫颈癌组织FIGO分期Ⅰ~Ⅱ期的VEGF阳性率、CD105-MVD低于Ⅲ~Ⅳ期,而PTEN阳性率高于Ⅲ~Ⅳ期;宫颈癌组织伴有淋巴结转移的VEGF阳性率、CD105-MVD高于无转移,而PTEN阳性率低于无转移,差异均有统计学意义(P0.05);Sperman相关性显示VEGF与PTEN,CD105-MVD与PTEN呈现明显的负相关关系(rs=-0.564,-0.893,P0.05),VEGF与CD105-MVD呈现正相关关系(rs=0.825,P0.05)。结论:PTEN在宫颈癌组织表达降低,CD105和VEGF升高,三个指标在宫颈癌的发生、发展、转移及预后中具有重要作用。     

彩色能量多普勒超声阻力指数与宫颈癌患者VEGF、MVD以及临床病理特征的关系

目的:研究彩色能量多普勒超声阻力指数与宫颈癌患者血管内皮生长因子(VEGF)、微血管密度(MVD)以及临床病理特征的关系。方法:以2015年2月-2018年2月我院收治的宫颈癌患者50例为研究对象,记为宫颈癌组。另取同期于我院接受治疗的宫颈上皮内瘤变患者50例记为宫颈上皮内瘤变组,取同期于我院体检的健康志愿者50例记为对照组。采用经阴道彩色能量多普勒超声检测三组受试者阻力指数,采用免疫组织化学法检测三组受试者VEGF表达以及MVD值。分别比较三组受试者阻力指数、VEGF表达及MVD值,并分析宫颈癌患者阻力指数与临床病理特征的关系,同时采用Spearman相关性分析宫颈癌患者阻力指数与VEGF、MVD的相关性。结果:宫颈癌组阻力指数低于宫颈上皮内瘤变组和对照组,且宫颈上皮内瘤变组低于对照组(P0.05);而宫颈癌组VEGF表达与MVD值高于宫颈上皮内瘤变组和对照组,且宫颈上皮内瘤变组高于对照组(P0.05)。病灶直径40 mm、临床分期Ⅰ-Ⅱ期、中高分化、无淋巴结转移宫颈癌患者阻力指数均高于病灶直径≥40 mm、临床分期Ⅲ-Ⅳ期、低分化、有淋巴结转移宫颈癌患者(P0.05),而阻力指数与宫颈前后径、组织学类型无关(P0.05)。经Spearman相关性分析可得:宫颈癌患者阻力指数与VEGF以及MVD均呈负相关(P0.05)。结论:彩色能量多普勒超声可有效反映宫颈病变形态结构特征、血管生成状况以及浸润情况,临床工作中可联合彩色能量多普勒超声与VEGF、MVD的测定,有助于判断宫颈癌患者病情严重程度,为临床治疗方案的制定以及预后评估提供参考依据。     

VEGF-D和CD105在胰腺癌中的表达及意义

目的:探讨VEGF-D,CD105在胰腺癌发生、发展及转移过程中的意义.方法:采用免疫组织化学S-P法,检测VEGF-D,CD105在53例胰腺癌组织(胰腺癌组)及10例正常胰腺组织(对照组)中的表达,计数MVD,并与临床病理学资料进行分析比较.结果:胰腺癌组织中VEGF-D的阳性表达明显高于正常胰腺组织(P＜0.05).VEGF-D的表达与胰腺癌的临床分期及淋巴结有无转移有关(P＜0.05).胰腺癌中,CD105标记的微血管密度(MVD)明显高于非肿瘤性胰腺组织.CD105的表达水平与胰腺癌临床分期及病理分级有关.胰腺癌组织中CD105抗体标记的微血管密度(MVD)结果显示:在有淋巴结转移的癌组织中,其MVD值高于无淋巴结转移组有显著统计学意义(P＜0.05).VEGF-D阳性表达组的MVD显著高于VEGF-D表达阴性组的MVD(P＜0.05):CD105在胰腺癌中的表达与VEGF-D的表达呈正相关(P＜0.05).结论:VEGF-D的表达与胰腺癌的血管生成有关.MVD值有可能作为判断胰腺癌生物学行为的潜在指标.     

胃癌及相应癌旁组织中VEGF、CD34、CD44v6的检测及临床意义

目的:探讨血管内皮生长因子(Vascular endothelial growth factor,VEGF)、CD34和CD44v6在胃癌及相应癌旁组织中的表达及其与临床病理意义.方法:应用免疫组化技术检测60例胃癌以及相应癌旁组织中VEGF、CD34和CD44v6的表达.结果:胃癌组织VEGF阳性率为71.67%(43/60)明显低于癌旁组织88.34%(53/60),两组间有显著性差异(P=0.022);VEGF阳性表达与胃癌病理分级、浸润深度、淋巴结转移和临床分期密切相关(P＜0.05).癌旁组织中微血管密度(MVD)明显高于癌组织MVD(P=0.000),有淋巴结转移癌组织中MVD高于无淋巴结转移者(P=0.043),并随浸润深度、临床分期MVD升高(P=0.046,P=0.000).癌旁组织中CD44v6阳性率为48.34%(29/60)低于癌组织的61.67%(37/60),两者无明显差别.CD44v6的阳性率随胃癌浸润的加深而升高,与淋巴结转移和TNM分期呈正相关.VEGF、CD34及CD44v6三指标间两两相关(P＜0.05).结论:VEGF、CD34和CD44v6三者联合检测有助于判断胃癌的浸润、转移及预后情况.     

术前介入栓塞化疗对局部晚期宫颈癌的疗效及VEGF表达的影响

目的:探讨局部晚期宫颈癌术前介入化疗的疗效及对VEGF的表达。方法:评定47例局部晚期宫颈癌介入化疗效果并应用RT-PCR方法测定化疗前后组织中VEGF表达化疗近期。结果:介入栓塞化疗的总有效率87.2%,ⅠB2期、ⅡA期、ⅡB期宫颈癌介入化疗效果无明显差异(P>0.05),ⅢA期及ⅢB期宫颈癌介入治疗效果无明显差异(P>0.05)且治疗有效率明显低于前三组(P<0.05),治疗前局部晚期宫颈癌组织中VEGF的表达与肿瘤的临床分期、淋巴结转移、病理类型、组织学分级均明显相关(P<0.05);介入治疗后高分化宫颈癌及无淋巴结转移的宫颈癌中VEGF表达无明显下降(P>0.05);不同临床分期、有淋巴结转移、不同病理类型及中、低分化的局部晚期宫颈癌介入治疗后VEGF表达均明显降低(P<0.05)。结论:术前介入栓塞化疗对局部晚期宫颈癌有良好近期效果,是一种可行、有效的辅助治疗手段。VEGF可作为判断治疗效果的指标。     

紫杉醇联合奈达铂新辅助治疗局部晚期宫颈癌的临床效果研究

目的:探讨紫杉醇联合奈达铂治疗局部晚期宫颈癌患者的近远期疗效。方法:选择2010年3月～2011年8月在我院进行诊治的局部晚期宫颈癌患者67例,按住院序号分为观察组33例和对照组34例。对照组单纯给予腹腔镜宫颈癌根治手术治疗,观察组联合给予新辅助化疗(静脉注射紫杉醇135 mg/m2以及奈达铂75 mg/m2)。比较两组的临床治疗效果,术后的宫旁累及阳性率、阴道切缘阳性率、淋巴结转移阳性率以及脉管浸润阳性率。对两组患者进行3年的随访,比较两组的复发率、转移率以及3年生存率。结果:观察组的近期治疗有效率为93.94%(31/33),明显高于对照组的70.59%(24/34)(P0.05);与对照组相比,观察组术后宫旁累及阳性率、阴道切缘阳性率、淋巴结转移阳性率、脉管浸润阳性率均明显较低(P0.05);经过3年的随访,观察组的复发率以及转移率明显低于对照组(P0.05),3年生存率明显高于对照组(P0.05)。结论:紫杉醇联合奈达铂新辅助治疗局部晚期宫颈癌的近期疗效以及远期疗效明显优于单纯给予腹腔镜宫颈癌根治手术治疗。     

姜黄素对晚期宫颈癌患者Bcl-xL、Bcl-2、EphA2及EphrinA1表达的影响

目的:探究姜黄素对宫颈癌晚期放疗患者Bcl-x L、Bcl-2、Eph A2与Ephrin A1表达的影响。方法:收集160例晚期宫颈癌放疗患者的宫颈癌石蜡标本,分为姜黄素组与对照组,均给予放射治疗,姜黄素组放疗期间加服姜黄素片剂,采用免疫组化等实验方法,观察两组病例标本Bcl-x L、Bcl-2、Eph A2与Ephrin A1的表达情况,比较两组病理标本化疗前后的凋亡细胞指数(AI)与微血管密度(MVD)。结果:姜黄素组的Bcl-x L、Bcl-2、Eph A2与Ephrin A1的表达阳性率分别为8.3%、43.33%、46.67%、15.0%,均显著低于对照组(P0.05);放疗后姜黄素组的AI显著高于对照组(P0.05),MVD显著低于对照组(P0.05)。结论:姜黄素可抑制Bcl-x L、Bcl-2的表达以促进肿瘤细胞凋亡,同时降低Eph A2与Ephrin A1的表达以减少肿瘤微血管形成,有显著的抗肿瘤与放疗增敏作用。     

食管鳞状细胞癌中HDGF、VEGF表达与微血管形成的关系

目的:研究食管鳞状细胞癌中肝癌衍生生长因子(HDGF)、血管内皮生长因子(VEGF)的表达及其与微血管形成的关系。方法:通过免疫组化SABC法检测和比较68例食管鳞癌、20例切缘正常组织中HDGF、VEGF的表达和CD34标记的微血管密度(MVD),分析HDGF和VEGF表达之间的关系及其与食管鳞癌患者临床病理因素和食管癌组织MVD值的关系。结果:食管鳞癌组织中HDGF(63.2%)和VEGF(72.1%)的阳性表达率均明显高于切缘正常粘膜组织(15.0%、20.0%)(P0.05),食管鳞癌组织和切缘正常粘膜组织中的MVD值分别为35.48±5.75和13.50±2.1(P0.05)。食管鳞癌组织HDGF的阳性表达率仅与其临床分期明显相关(P0.05),而VEGF的阳性表达率与其淋巴结转移、临床分期均显著相关(P0.05),二者在食管鳞癌组织中的表达呈显著正相关(P0.05)。食管鳞癌组织中HDGF、VEGF阳性表达组MVD值均明显高于HDGF、VEGF阴性表达组(P0.05)。结论:HDGF可能通过诱导VEGF的产生,从而促进血管生成,参与食管鳞癌的发生、发展及转移。     

PF化疗同步联合时辰放疗与常规放疗治疗局部晚期鼻咽癌的随机对照研究

目的:比较局部晚期鼻咽癌分别接受顺铂联合氟尿嘧啶方案(PF)化疗联合时辰放疗以及PF化疗联合常规放疗时的疗效和毒副反应。方法:将我科45例初治局部晚期鼻咽癌患者随机分为时辰放疗组和常规放疗组,分别为22例和23例。两组均采用相同放射治疗方法和治疗剂量,同步给予PF方案化疗。时辰放疗组放疗时间在20:00-22:00,而常规放疗组在8:00-10:00。结果:两组患者在放疗结束3个月和6个月进行时比较发现,时辰放疗组患者的鼻咽部及颈部的肿瘤完全消退率(CR率)均比常规放疗组高(P<0.05)。而时辰放疗组的各种急性副作用发生率较常规放疗组低,但无统计学差异。治疗后时辰组CD4/CD8升高,常规组CD4/CD8降低(P<0.05)。结论:时辰放疗联合PF同步化疗针对局部晚期鼻咽癌患者疗效好,毒副作用轻,同时可能有利于改善患者的免疫功能。     

血管紧张素Ⅱ受体1、VEGF和MVD在乳腺癌中的表达及相关性研究

目的:研究AT1受体、VEGF和CD34在乳腺癌中的表达、并探讨其相关性及临床意义。方法:该实验采用免疫组化方法(SP法),对102例乳腺癌患者石蜡包埋切片中的AT1受体、VEGF和CD34进行检测。结果:AT1受体在乳腺癌中阳性表达率为48%,在乳腺癌中有腋淋巴结转移组为61．8%,无腋淋巴结转移组为31．9%,两者差别有统计学意义(P＜0．05)。VEGF在乳腺癌中表达率为40．2%,有腋淋巴结转移组44．4%,无腋淋巴结转移组为23．5%,两者差别有统计学意义(P＜0．01)。AT1R阴性表达组的VEGF阳性表达率为11．3%(6/53),AT1R阳性表达组的VEGF阳性表达为71．4%(35/49),两者差别有统计学意义(P＜0．05); AT1R阴性表达组的MVD值为17．35±5．67,AT1R阳性表达组的MVD值为20．37±7．30,两组差别有显著性(P＜0．05)。VEGF阴性表达组的MVD表达率为17．14±5．78,VEGF阳性表达组的MVD表达为21．27±7．14,两组差别有统计学意义(P＜0．05)。结论:乳腺癌组织中AT1受体的阳性表达率为48%(49/102)AT1受体表达与患者年龄、肿瘤大小、ER、PR无相关性,而与腋淋巴结转移成正相关;乳腺癌组织中VEGF的阳性表达率为40．2%(41/102),VEGF表达与患者年龄、肿瘤大小、ER、PR无相关性,而与腋淋巴结转移成正相关;AT1受体表达与VEGF表达成正相关。在乳腺癌组织中MVD和AT1R和VEGF表达均成正相关。     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

放牧对牧草光合作用、呼吸作用和氮、碳吸收与转运的影响

研究放牧对草地植物生理活动的影响,对于揭示草地放牧演替的生理机制有重要意义.大量研究表明,家畜放牧对牧草光合作用、呼吸作用以及C和N吸收与转运的影响,可以分为生理伤害和生理恢复2个阶段.放牧通过改变草地冠层结构影响牧草光合作用,净光合作用速率短期内迅速下降,随着叶面积指数增加又逐渐上升,呼吸作用有相似的变化趋势.牧草放牧后再生长所需的C和N最初主要来自根系和留茬中的贮藏物质,此后随着牧草生长恢复逐渐由同化作用供给,C代谢与土壤N水平负相关.放牧后牧草生理活动变化与牧草遗传特性、种间竞争、家畜放牧特征、非生物环境等因素密切相关.     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000     

Coiled-coil nanomechanics and uncoiling and unfolding of the superhelix and alpha-helices of myosin

The nanomechanical properties of the coiled-coils of myosin are fundamentally important in understanding muscle assembly and contraction. Force spectra of single molecules of double-headed myosin, single-headed myosin, and coiled-coil tail fragments were acquired with an atomic force microscope and displayed characteristic triphasic force-distance responses to stretch: a rise phase (R) and a plateau phase (P) and an exponential phase (E). The R and P phases arise mainly from the stretching of the coiled-coils, with the hinge region being the main contributor to the rise phase at low force. Only the E phase was analyzable by the worm-like chain model of polymer elasticity. Restrained molecular mechanics simulations on an existing x-ray structure of scallop S2 yielded force spectra with either two or three phases, depending on the mode of stretch. It revealed that coiled-coil chains separate completely near the end of the P phase and the stretching of the unfolded chains gives rise to the E phase. Extensive conformational searching yielded a P phase force near 40 pN that agreed well with the experimental value. We suggest that the flexible and elastic S2 region, particularly the hinge region, may undergo force-induced unfolding and extend reversibly during actomyosin powerstroke.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。     

Synthesis of hapten and conjugates of coumestrol and development of immunoassay

3-O-Carboxymethylcoumestrol was prepared as the hapten for immunoassay by a partial alkylation of coumestrol with ethyl chloroacetate in acetone alkalized with potassium carbonate. 3-O-Ethoxycarbonylmethylcoumestrol was separated by column chromatography and finally was hydrolyzed with formic acid. 1H and 13C NMR data (APT, COSY, HMQC, and HMBC) revealed that the reaction was regioselective, as 3-O-ethoxycarboxymethylcoumestrol was the only monosubstituted derivative. The hapten was then conjugated to bovine serum albumin and used for immunization of rabbits. A radioimmunoassay (RIA) system was established based on the polyclonal antiserum and a 125I-labeled hapten-tyrosine methyl ester conjugate as the radioligand. Parameters of the RIA: sensitivity: 12 pg per tube, 50% intercept: 140 pg per tube, working range: 20-4000 pg per tube. The cross-reactivity of a panel isoflavonoid and lignan phytoestrogens was either negligible (e.g. formononetin 0.07%; biochanin A 0.06%) or not detectable at all. The major immunoreactive peak in HPLC fractions from an alfalfa extract had the same retention time as coumestrol standard and represented 94.8% of the signal. The remaining 5.2% of immunoreactivity was distributed between five minor peaks. We conclude that after the validation for particular matrices, the method will be a useful tool for analysis of coumestrol, especially in low volume and low concentration samples.     

Morphology and putative function of the colon and cloaca of marine and freshwater snakes

Among tetrapods, evidence for postrenal modification of the urine by the distal digestive tract (including the colon and cloaca) is highly variable. Birds and bladderless reptiles are of interest because the colon and cloaca represent the only sites from which water and ions can be reclaimed from the urine secreted by the kidney. For animals occupying desiccating environments (e.g., deserts and marine environments), postrenal modification of the urine may directly contribute to the maintenance of hypo‐osmotic body fluids. We compared the morphology and distribution of key proteins in the colon, cloaca, and urogenital ducts of watersnakes from marine (Nerodia clarkii clarkii) and freshwater (Nerodia fasciata) habitats. Specifically, we examined the epithelia of each tissue for evidence of mucus production by examining the distribution of mucopolysaccharides, and for evidence of water/ion regulation by examining the distribution of Na⁺/K⁺‐ATPase (NKA), Na⁺/K⁺/Cl^? cotransporter (NKCC), and aquaporin 3 (AQP3). NKCC localized to the basolateral epithelium of the colon, urodeal sphincter, and proctodeum, consistent with a role in secretion of Na⁺, Cl^?, and K⁺ from the tissue, but NKA was not detected in the colon or any compartment of the cloaca. Interestingly, NKA was detected in the basolateral epithelium of the ureters, suggesting the urothelium may play a role in active ion transport. AQP3 was detected in the ureters and coprodeal complex, consistent with a role in urinary and fecal dehydration or, potentially, in the production of the watery component of the mucus secreted by the coprodeal complex. Since no differences in general cloacal morphology, production of mucus, or the distribution of ion transporters/water channels were detected between the two species, cloacal osmoregulation may either be regulated by proteins not examined in this study or may not be responsible for the differential success of N. c. clarkii and N. fasciata in marine habitats. J. Morphol. 2011. © 2011 Wiley Periodicals, Inc.