Abnormal Secondary Growth and Histochemical Localization of Alkaloids in Root System of Aconitum flavum Hand.-Mazz.

[Objective] The purpose of this study was to clarify the structure,growth pattern and histochemical localization of alkaloids in root system of Aconitum flavum Hand.-Mazz.[Method] Paraffin sectioning and histochemistry were employed for performing the analysis in this study.[Result] The root system of Aconitum flavum Hand.-Mazz.consists of taproot,lateral root and adventitious root.The primary structure of root system is normal,but secondary structure shows abnormal.The cambium and the extra cambium of taproot form a ＂U＂-shaped secondary vascular bundle and tertiary bundle in abnormal secondary structure.The sieve tube group is made of little sieve tube group which is differentiated from primary phloem and cambium.Meanwhile,the secondary xylem in tuberous root also appears to be a ＂U＂ shape.Parenchyma cells of secondary phloem occupy most of the tuberous root.The sieve tube group of tuberous root is mainly differentiated from parenchyma cell of secondary phloem.[Conclusion] The difference in abnormal secondary structure of taproot and tuberous root are attributed to their varied cambium compose and activity pattern.Alkaloids are mainly accumulated in parenchyma cell of the inside cortex and between bundle in taproot,while parenchyma of secondary phloem and pith in tuberous root.

Dynamical changing patterns of glycogen and enzyme histochemical activities in rat liver graft undergoing warm ischemia injury

AIM:To investigate the changing patterns of glycogen and enzyme histochemical activities in rat liver graft under a different warm ischemia time (WIT) and to predict the tolerant time limitation of the liver graft to warm ischemia injury. METHODS: The rats were randomized into five groups, WIT was 0,15,30,45,60 min, respectively, and histochemical staining of liver graft specimens was observed. The recovery changes of glycogen and enzyme histochemistry activities were measured respectively 6 and 24 h following liver graft implantation. RESULTS: The activities of succinic dehydrogenase, cytochrome oxidase, apyrase (Mg++-ATPase) and content of glycogen were decreased gradually after different WIT in a time-dependent manner. The changes were significant when WIT was over 30 min. CONCLUSION: Hepatic injury is reversible within 30 min of warm ischemia injury. Glycogen and enzyme histochemistry activities of liver grafts and their recovery potency after reperfusion may serve as criteria to evaluate the quality of liver grafts.

Immunohistochemical Analysis of the Acid Secretion Potency in Gastric Parietal Cells

Gastric parietal cells are important in acid secretion, but it is unclear which cells throughout the gastric gland have the highest secretion potency. Here, we used immunohistochemical methods with anti-H+, K+-ATPase, phosphoryl ezrin and CD44 antibodies to study the distribution of gastric acid secretion activity. Stomach tissues from freely fed and starved rats were cryofixed for light microscopy or fixed by high-pressure freezing for electron microscopy. Parietal cells from freely fed animals corresponded to the active secretion phase and to the inactive resting phase from starved rats. Anti-H+, K+-ATPase and anti-phosphoryl ezrin labeling were observed on the membrane of the intracellular canaliculi and the tubulovesicle from freely fed rats, while cells from starved animals showed weak labeling with anti-phosphoryl ezrin antibody staining. Morphometrical analysis at the electron microscopic level was performed on active and inactive acid secretory phases between the upper and base regions of the gland. H+, K+-ATPase and CD44 were distributed on both sites of the microvillous and tubulovesicle membrane in the same cells, but phosphoryl ezrin localized predominantly on the microvillous membrane in active cells of the glandular neck and upper base. Therefore, the highest secreting potency appeared to be in cells of the glandular neck and upper base.

HISTOCHEMICAL AND IMMUNOHISTOCHEMICAL STUDY OF EXTRAMAMMARY PACET'S DISEASE

The histogensis of extramammary Paget's disease has not been solved and remained controversial. Eight cases of extramammary Paget's disease of geni-tocrural region were investigated by alcian blue and PAS stain and immunoreaction of anti-CEA and anti-keratin. It was found that the pattern and intensity of alcian blue, PAS staining were identical for Paget cells and secretory cells of apocrine sweat gland; and CEA immunoreactivity was uniformly observed in both Paget cells and eccrine sweat gland. The keratin immunoreaction was positive in keratinocytes, apocrine and eccrine sweat gland, whereas Paget cells were negative. These results suggested that Paget cells of extramammary Paget's disease could be derived from multipotential epidermal germ cells.

HISTOCHEMICAL EFFECT OF THE MULTIGLYCOSIDES OF TRIPTERYGIUM WILFORDII HOOK.F.(GTW)ON THE TESTIS AND EPIDIDYMIS OF RATS

SD Twenty adult male rats were equally divided into a contro1 and an experimcntal group.GTW,10 mg/kg per day,was given to the experimental rats through gastric gavage 6 days a week for 8 weeks.To the control rats,the same amount of vehicle was given The animals were sacrificed when they became infertile and the fol1owing parameters of the testis and epididymis were examined:l)DNA(Feulgen′s method),2)RNA(Brachet′s tech-uique),3)LDH-X(Lojda′s method),4)ATPase(Wachstein′s procedure),5)Succinate dehydrc-genase(SDH,Pearson's method),6)Non-specific esterase(NSE,Lojda's technique)and 7)PAS reaction.Routine examination of the epididymal spermatozoa was also carried out.

Anatomy, Histochemistry and Phytochemistry of Leaves in Aloe vera var. chinensis

A multidisciplinary approach-anatomy, histochemistry and phytochemistry-was used to investigate the leaf structure, the content and the storage location of barbaloin in the leaves of Aloe vera L. var. chinensis (Haw.) Berg. Xeromorphic characteristics including secondary thickened epidermal cell walls, thicker cuticle, ambiguous differentiation of spongy and palisade tissues in the chlorenchyma, and well-developed aquiferous tissue could be seen in the leaves. Several large parenchymatous cells were observed at the phloem pole of the first ring of vascular bundles. The secondary ring of vascular bundles in the leaf base and the stomata, which are surrounded by five cells, have some classification significance in this species. The density of vascular bundles, the content of barbaloin and the intensity of histochemical reaction differed among leaf numbers Ll (annual leaf), L2 (biennial leaf), L3 (triennial leaf) and L4 (quadrennial leaf), and in different parts of the leaf. These three factors were highest in the youngest leaf, Ll, and top parts of all the leaves and lowest in the basal parts and the oldest leaf, L4. The density of vascular bundles had a positive correlation to the content of barbaloin. The histochemical results revealed that the small sheath cells that surrounded the bundles might be the location of barbaloin synthesis and the large parenchymatous cells beneath the sheath might be the storage places of this metabolite.

THE ULTRASTRUCTURE AND ENZYME HISTOCHEMISTRY OF THE SUBSTANTIA GELATINOSA OF RAT SPINAL CORD UNDER THE INFLUENCE OF ELECTRO-ACUPUNCTURE ANALGESIA

Histochemistry and EM were used in this experiment to study the effect of EAon the spinal dorsal horn of rat. After 30 minutes EA, the pain threshold of EA rats were elevated400%, the acid phosphatase reaction in spinal dorsal horn of EA rats increased obviously, the roundagranular vesicles of the C-terminals of EA group was more than that of control’s. The results sug-gested that the histochemical components and numbers of synaptic vesicles in SG and their functionscould be regulated by acupuncture.

不同级别胶质瘤患者MEK、ERK、CHKα蛋白的阳性表达情况及临床意义

目的探讨不同级别胶质瘤患者胆碱激酶α(CHKα)、细胞外信号调节激酶(ERK)及丝裂原激活蛋白激酶(MEK)蛋白的阳性表达情况及临床意义。方法回顾性分析2019年1月至2022年12月长沙市第四医院神经外科收治的124例胶质瘤患者的临床资料。检测、统计不同级别胶质瘤患者MEK、ERK、CHKα蛋白的阳性表达情况。结果不同肿瘤最大径、年龄、WHO分级及预后胶质瘤患者MEK、ERE、CHKα蛋白的高、低表达情况比较,差异均有统计学意义(P<0.05)。Ⅰ、Ⅱ、Ⅲ、Ⅳ级胶质瘤患者MEK蛋白阳性表达率分别为9.62%、7.69%、94.08%和92.04%,ERK蛋白阳性表达率分别为9.57%、7.49%、94.52%和92.18%,CHKα蛋白阳性表达率分别为9.60%、7.53%、94.71%和92.23%,高、低级别胶质瘤患者MEK、ERK、CHKα蛋白阳性表达率比较,差异均有统计学意义(P<0.05)。随访结束后结果显示,124例患者中死亡52例,失访12例,获得随访112例,随访率为90.32%。结论不同级别胶质瘤患者MEK、ERK、CHKα蛋白的阳性表达情况有明显区别,并且与患者的疾病预后明显相关,对疾病的诊断和预后判断具有重要价值。

两栖植物香附子的解剖结构和组织化学研究

以分布在三峡水库消落区和江汉平原河漫滩香附子营养器官为试验材料,采用解剖镜下切片法、组织化学研究方法、光学和荧光显微镜下观察拍照记录试验结果,研究其叶、直立茎、根状茎、块茎和不定根适应两栖环境的解剖结构与组织化学特征。结果表明:1)香附子具有适应两栖环境的气腔和质外体屏障等典型结构特征,其中气腔包含根、根状茎及叶的溶生性通气组织和茎的裂生性通气组织;质外体屏障包括内皮层、外皮层、栓质化中柱和维管束鞘细胞等。2)香附子的不定根和茎具有栓质化中柱和维管束;叶片和茎的花环结构由内侧维管束鞘、中间维管束鞘和薄壁细胞维管束鞘3层组成。3)香附子的质外体屏障可能有助于香附子在淹没缺氧下保持氧气流通,同时控制水、离子与环境交换,气腔有利于保存和输送有氧呼吸所必需的氧气。香附子适应淹没-陆生两栖转换环境的结构特征表明其是长江流域生态恢复的重要植物资源。

冷诱导RNA结合蛋白联合IL-6的检测在妊娠期高血压疾病中的诊断价值

目的探讨冷诱导RNA结合蛋白(CIRBP)联合白细胞介素-6(IL-6)检测在妊娠期高血压疾病中的诊断价值。方法前瞻性选取2022年7月至2023年10月入上海市第四人民医院妇产科病区经剖宫产分娩的高血压产妇30例作为实验组,选取正常产妇30例作为对照组,分别采用免疫组织化学染色、Western blot、qRT-PCR和ELISA法检测两组研究对象胎盘组织及血浆中CIRBP、IL-6的表达并进行对比分析。根据疾病程度将实验组患者分为妊娠期高血压组(12例)和重度子痫前期(18例)。比较两组患者的血浆中CIRBP和IL-6的表达水平。采用ROC曲线分析CIRBP+IL-6联合检测对疾病的诊断效能。结果实验组患者胎盘组织中CIRBP和IL-6阳性表达率明显高于对照组(P<0.05);实验组患者胎盘组织中CIRBP和IL-6 mRNA和蛋白表达明显高于对照组(P<0.05);实验组患者中血浆中CIRBP和IL-6表达明显高于对照组(P<0.05);重度子痫前期组患者血浆中CIRBP和IL-6表达明显高于妊娠期高血压组(P<0.05)。结论妊娠高血压患者胎盘组织及血浆中CIRBP和IL-6均呈高表达,且随着病程的加重不断升高,对诊断高血压产妇有深远的影响。

番茄磷酸盐转运蛋白SlPT2基因启动子克隆及功能分析

为研究番茄磷酸盐转运蛋白SlPT2基因启动子的功能及活性,以番茄Micro-Tom为材料,利用荧光定量PCR检测SlPT2基因表达模式,利用PCR技术克隆SlPT2启动子(p SlPT2)序列,通过启动子在线分析网站Plant Care和New Place分析p SlPT2内部所含有的顺式作用元件,利用同源重组法构建p1300GN-p SlPT2重组载体,转化拟南芥,通过GUS组织化学染色研究SlPT2启动子表达模式及表达活性。荧光定量PCR结果表明,SlPT2主要在番茄根部表达。通过Plant Care和New Place分析发现,启动子(1 801 bp)内部除含有CAAT-Box与TATA-Box等核心启动子元件外,还包含与根特异性表达有关的元件(ROOTMOTIFTAPOX1、RAV1AAT、OSE1ROOTNODULE),以及参与光响应(Box 4、G-Box、TCTmotif)、茉莉酸甲酯反应(CGTCA-motif)、脱落酸反应(ABRE)、玉米醇溶蛋白代谢调控(O2-site)的顺式作用元件等。GUS组织化学染色结果表明,SlPT2启动子驱动GUS基因在拟南芥根部特异表达,将其初步鉴定为根特异性表达启动子。

间接免疫组化法检测鸭疫里默氏杆菌感染和抗原定位

应用鸭源血清1型鸭疫里默氏杆菌(RA)作为抗原,免疫兔制备兔抗RA的IgG,建立了检测雏鸭感染鸭疫里默氏杆菌的间接免疫酶组织化学法。该法检测大肠杆菌(O1、O8、O79、O138)、鸭沙门氏菌、多杀性巴氏杆菌(5∶A)感染发病死亡雏鸭组织,不出现阳性反应,而检测RA感染发病死亡雏鸭组织出现特异性阳性反应;检测RA人工发病死亡雏鸭,可在心脏、肝脏、肺脏、肾脏、十二指肠、盲肠、直肠、脾脏、法氏囊、胸腺、胰脏、脑和腺胃检测到RA抗原,RA抗原分布于细胞浆、组织间隙和血液;检测RA人工感染病例与RA肝脏分离符合率为100%,检测临床可疑病例的血清1型RA的阳性率(92.96%)比细菌分离率(75.12%)高。该法具有特异、直观和敏感的特点,可用于雏鸭RA人工感染和临床感染的诊断、检测及RA抗原定位和RA致病机理的研究。

日本血吸虫培养细胞SDH、AKP和ACP的组织化学观察

目的 探讨日本血吸虫成虫培养细胞琥珀酸脱氢酶 (SDH)、碱性磷酸酶 (AKP)和酸性磷酸酶 (ACP)的组织化学变化。方法 应用组织化学染色方法显示日本血吸虫成虫培养细胞SDH、AKP和ACP。结果 日本血吸虫成虫培养细胞均具SDH、AKP、ACP活性 ,体积较大的细胞在胞膜附近和鞭毛细胞的鞭毛部位SDH活性最强 ;圆颗粒细胞、三角扇形细胞及合胞体的AKP活性细胞质较细胞核弱 ,团簇状排列的细胞主要分布在细胞质 ,而鞭毛形细胞则集中于细胞核和鞭毛部位 ;ACP的活性均较强 ,尤其是体积较大的培养细胞 ,活性部位集中分布在核周或弥散分布在细胞质中。结论 日本血吸虫培养细胞SDH、AKP和ACP的组化染色可做为简便易行、快速敏感的细胞培养条件优劣的评价指标。

弹力纤维染色在评估肺癌胸膜侵犯中的意义

目的探讨胸膜弹力纤维(elastic fibers,EFs)染色在肺癌TNM分期应用中的价值。方法随机选取手术切除的肺癌标本200例,肉眼观察癌组织均紧贴胸膜。石蜡切片,行HE及EFs染色。按照WHO(2004)肺癌分类标准,对200例标本进行组织学类型分类、观察并分析HE及EFs染色结果。结果光镜下对200例标本进行组织学分类,其中162例为腺癌、29例为鳞癌、4例为腺鳞癌、5例为大细胞癌;其中肿瘤细胞未突破胸膜弹力层92例,突破胸膜弹力层108例(含破脏层胸膜20例);HE染色28例无法判断或判断不清,最终经EFs染色得出结论。结论应用胸膜EFs染色,能直观地观察肿瘤细胞是否到达或突破胸膜层,有助于准确评估肺癌的胸膜侵犯和TNM分期。

2种石斛属植物和小叶石仙桃多糖的组织化学定位

为对不同年龄的铁皮石斛(Dendrobium candidum)、铜皮石斛(D.moniliforme)及石斛伪品-小叶石仙桃(Pholidota chinensis)不同器官中的多糖进行组织化学定位,选择具有较高药用价值的石斛类药用植物和药用部位,确定铁皮石斛的最佳采收期,根据多糖的理化性质,材料经冰冻切片,用改良的高碘酸-席夫试剂(PAS)对多糖进行组织化学定位;结果表明,多糖类物质主要存在茎的薄壁细胞、根的皮层细胞、叶的叶肉细胞中。其总多糖含量有如下顺序:二年生的铁皮石斛茎>铜皮石斛茎≈>小叶石仙桃茎,铁皮石斛根≈铜皮石斛根>小叶石仙桃根,铁皮石斛叶≈小叶石仙桃叶>铜皮石斛叶。另外,在不同的器官中,铜皮石斛的多糖含量呈现“茎≈根>叶”和小叶石仙桃的多糖含量呈现“叶>茎≈>根”的规律,而二年生的铁皮石斛则为“茎>叶>根”。这表明在种间及同种植物的不同器官、不同年龄,其多糖的含量存在差异。而3种兰科植物中,以二年生铁皮石斛茎的多糖含量最高,表明其药用价值也最高。

芦荟属植物叶内蒽醌类物质的组织化学定位研究

蒽醌类物质是芦荟属 (Aloe)植物叶内贮存的重要次生代谢物质 ,主要是芦荟素 ,高那特芦荟素、芦荟苦素和芦荟宁等。根据蒽醌类物质的理化性质 ,用 2 %～ 5% Na OH和 5% Pb(CH3 COO) 2 溶液对材料进行处理 ,再辅以荧光和超薄切片观察等多种方法 ,对芦荟叶内的 4种主要蒽醌类物质进行了组织化学定位研究。结果表明 ,蒽醌类物质在芦荟叶内的贮藏是多位点的。芦荟素和高那特芦荟素主要位于韧皮部的大型薄壁组织细胞之中 ;而芦荟苦素和芦荟宁主要位于维管束鞘细胞以及贮水组织与同化薄壁组织之间的 1圈薄壁组织细胞之中 ,贮水组织细胞中不贮存蒽醌类物质。

金钗石斛生物碱的组织化学定位研究

利用组织化学定位方法对金钗石斛不同生长年限和不同营养器官的生物碱分布和积累进行比较分析。结果表明,金钗石斛茎生物碱主要分布于基本组织,三年生金钗石斛的生物碱含量高于一年生和二年生金钗石斛。金钗石斛根部生物碱含量同样较多,主要分布于皮层薄壁细胞中;金钗石斛叶片生物碱含量较低,存在于上下表皮细胞中,与铁皮石斛和铜皮石斛完全不同。说明三年生金钗石斛作为最适宜采收期,而且金钗石斛根部具有很好的开发利用价值。金钗石斛叶片表皮细胞中呈现生物碱的原因尚未清楚,需要进一步研究。

冰冻切片技术在植物显微结构和组织化学中的应用

介绍了冰冻切片法研究植物显微结构和组织化学的一般程序。结果表明,冰冻切片过程简单有效且图版清晰,在1d内可获得高质量图片,解决了利用普通石蜡切片观察植物样品需要进行脱水、浸透、包埋操作且耗时长的问题,在植物显微结构和组织化学研究中具有广阔的应用前景。

二种石斛属植物和小叶石仙桃生物碱的组织化学定位

目的对不同年龄的铁皮石斛、铜皮石斛及其石斛伪品———小叶石仙桃的不同器官进行生物碱组织化学定位,选择具有较高药用价值的石斛类药用植物和药用部位,确定铁皮石斛的最佳采收期。方法采用冰冻切片法,根据生物碱的理化性质,用碘化铋钾染色法对3种植物的生物碱进行组织化学定位。结果生物碱主要存在于3种植物茎的薄壁细胞、根的皮层细胞、叶的叶肉细胞中;其总生物碱含量顺序为:二生年的铁皮石斛≈铜皮石斛>小叶石仙桃。此外,在3种植物不同的器官中,其总生物碱的含量都呈现“叶>茎≈根”的规律。结论在种间及同种植物的不同年龄或不同器官中,其生物碱的含量存在差异。其中,以二年生铁皮石斛和铜皮石斛茎叶的生物碱含量较高,表明其药用价值也较高。

二甲基联苯胺检测植物叶片中H_2O_2方法的改良

对原二甲基联苯胺(3,3-diaminobenzidine,DAB)为染色剂检测植物叶片H2O2的组织化学方法进行改良研究。结果表明,与原方法相比,改良后的方法主要是在染色后不用体积分数为95%乙醇脱色,而用2%戊二醛+4%多聚甲醛溶液对1cm2左右叶片片段进行固定,固定24h后用冰冻切片机切成12μm薄片,显微镜下观察并拍照。用改良DAB方法对玉米叶片进行观察,结果显示,在水分胁迫4h时,玉米叶片的主脉及叶肉细胞叶绿体上均可观察到H2O2的积累,而原DAB方法观察不到叶肉细胞叶绿体上的H2O2积累。进一步用CeCl3染色的细胞化学方法验证,其结果与改良后的组织化学方法研究结果一致。研究表明,改良后的组织化学方法优于原方法,而且对植物组织中H2O2的化学定位可靠。