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.

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.

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 studies of distribution of acetylcholinesterase-positive fibers and peptidergic terminals in the nasal mucosa of rats

Objective To further investigate the mechanism of nasal secretion closely related to the innervation patterns in nasal mucosa with emphasis on the acetylcholinesterase (AChE) positive fibers and peptidergic terminals in nasal mucosa as well as trigeminal ganglion (TG) cells. Methods Histochemical demonstration of AChE positive fibers, immunohistochemical study of the distribution patterns of multiple peptidergic terminals, double labelling of AChE and substance P (SP) and somatostatin (SOM) mRNA in situ hybridization were carried out in nasal mucosa and trigeminal ganglion (TG) in rats. Results AChE positive terminals were mainly distributed in the mid to posterior one third of septal nasal mucosa, with greater staining density on the walls of small vessels and glands. There were fewer such terminals in turbinate mucosa. Tachykinins ergic terminals, including substance P(SP) , neurokinin A (NKA) , neurokinin B(NKB) and calcitonin gene related peptide (CGRP) ergic terminals, had an extensive localizations in nasal mucosa, involving the following areas: between epithelial cells, submucosa, the walls of small vessels, glands and venous sinusoids in both septal and turbinate nasal mucosa. Septal mucosa had the greater density. There were overlaps in the distribution of these peptidergic terminals. There were also vasoactive intestinal peptide (VIP) , neuropeptide Y (NPY) and galanin (GAL) ergic terminals in nasal mucosa. But no neurotensin (NT) and somatostatin (SOM) ergic terminals were found. In situ hybridization revealed SOMmRNA expression in TG cells. AChE and nine neuropeptides existed in the cytoplasms of TG cells. Besides, AChE and SP could exist simultaneously in cytoplasms of TG cells. Conclusions AChE positive (corresponding to parasympathetic nerves) and peptidergic terminals have different distribution patterns in the nasal mucosa of rats, although an overlap does exist, indicative of their different physiological effects on the regulation of nasal secretion and other functions; AChE and multiple neuropeptides in the cytoplasm of TG cells might play a role in modulating the nasal secretion in response to stimuli in the nasal mucosa.

LOCALIZATION OF THE SUPERIOR SALIVATORY NUCLEUS OF RAT——A STUDY USING COMBINATION OF AChE AND HRP HISTOCHEMICAL TECHNIQUES

Using acetylcholinesterase (AChE) histochemical technique, Shute and Lewis (1960) traced nervus intermedius, which innervated submandibular gland and sublingual gland, to two strongly AChE positive nuclei situated respectively at the medial and lateral sides of the genu of the facial nerve and regarded these two nuclei as the salivatory nuclei. However, recently, using retrograde transportation of horseradish pe-

Histopathological,histochemical and biochemical postmortem changes in induced fatal hypothermia in rats

Reaching a postmortem diagnosis of hypothermia is challenging in forensic practice.Therefore,this study was conducted to detect the histopathological,histochemical and biochemical changes that occur in adult albino rats following exposure to induced fatal hypothermia.Twenty-four adult albino rats were divided into the negative control,moderate hypothermia,severe hypothermia and hypoxia groups.Rats in the control group were euthanized when those in the moderate hypothermic group died.Blood samples were collected via heart puncture,and the cerebrum,heart,suprarenal gland,kidney,liver and skeletal muscle were removed to investigate the biochemical,histochemical and histopathological changes.Postmortem assessment depicted significant changes in lipid peroxidation,represented by increased malondialdehyde levels in the studied organs of the rats in hypothermic and hypoxia groups.Histopathological examination of the rats’organs revealed degeneration and necrosis in the hypothermia and hypoxia groups.Sections taken from the severe hypothermic rats revealed a loss of normal cardiac tissue architecture,necrotic changes in the pyramidal cells in the cerebral cortex,and massive necrosis,mainly in the tubules of the renal cortex and medulla.These findings suggest that histological changes might be used as biochemical markers for postmortem diagnosing of fatal hypothermia,particularly in severe hypothermic conditions.

Comparative histochemical and immunohistochemical study of benign and malignant colorectal tissues associated with Schistosoma japonicum infections

Abstract Objective Our previous studies have demonstrated a high percentage of mucinous carcinomas in colorectal cancer associated with schistosomiasis (CCS). This is a further study to explore whether there are special immunohistochemical and histochemical characters in CCS or its subtypes. Methods A comparative study was carried out in benign and malignant colorectal tissues with and without schistosomiasis. The study included a quantitative determination of peanut lectin (PNA) binding sites and proliferating cell nuclear antigen (PCNA) expression and histochemical detection of mucin changes. These methods have often been applied to the study of colorectal cancer but have not been recorded in the study of CCS. 133 cases were investigated, of which 70 cases were of CCS and 63 cases were of colorectal carcinoma without schistosomiasis (CC). The two groups were subdivided into mucin containing (MC) and non mucin containing (NMC) subtypes. Results Non tumorous mucosa of the CCS group showed 47% PNA positive cases, significantly higher than 25% in the CC group (P<0.01). The difference was mainly found in MC subtype of CCS. 65% of non tumorous mucosa adjacent to MC of the CCS group exhibited PNA binding, and in MC of the CC group, only 31% (P< 0.05). Moreover, the non tumorous mucosa in the cases of MC of the CCS group also showed a high percentage of sialo mucin predominant secretion (69% vs 38% in MC of the CC group, P<0.05).Department of Consistently, the presence of PNA binding sites in MC tumors of the CCS group was increased, compared with that in the same subtype in the CC group (65% and 31%, respectively, of strong positivity for PNA, P<0.05). However, no differences in expression of PNA and mucin changes were demonstrated in the surrounding mucosa and tumorous tissues of NMC between CCS and CC. The expression of PCNA was not different between CCS and CC and their subtypes. Conclusions Because PNA has an ability to bind to specific sugar molecules that have been proved to be closely related to early malignant change in colonic epithelia, and a predominance of sialomucins (50% or more) is regarded as “abnormal mucin pattern” valuable in identifying early development of epithelial dysplasia and pre malignant changes, the high frequency of PNA binding sites in non tumorous and tumorous tissues of the CCS group and its MC subtype and the increase of sialomucins in non tumorous mucosa of MC of the CCS group could be considered as a stronger tendency of malignant transformation in the colorectal epithelia influenced by schistosomiasis. If these malignant changes in the schistosomal colon are postulated to result in MC subtype tumors, it could explain the significantly high proportion of MC in the CCS group (P<0.05). With a review of current knowledge on the association of schistosomiasis and colorectal cancer, we propose that schistosomiasis japonica might have a close relation with mucin containing types of colorectal cancer.

Histochemical Localization of Nitric Oxide Synthase Positive Nerves in Chicken Oviduct

Nitric oxide(NO)is an important inhibitory neurotransmitter,synthesized by the enzyme nitric oxide synthase(NOS)locally in many tissues.The histochemical technique using nicotinamide adenine dinucleotide-diaphorase/nitric oxide synthase(NADPH-d/NOS)is often employed to determine the presence of this enzyme in the neuronal as well as other cell populations.We

Immunohistochemical classification and prognosis of diffuse large B-cell lymphoma in China

Objective To study the immunohistochemical classification and prognosis of diffuse large B-cell lymphoma(DLBCL).Methods A total of 148 cases of DLBCL were classified into germinal center B-cell-like(GCB)and non-GCB/activated B-cell-like(ABC)subtypes by Hans,Choi and Tally immunohistochemical stain algorithms.The clinical features and survival data of

Invasive breast carcinoma with osteoclast-like stromal giant cells:A case report

BACKGROUND Invasive breast carcinoma with osteoclast-like stromal giant cells(OGCs) is an extremely rare morphology of breast carcinomas.To the best of our knowledge,the most recent case report describing this rare pathology was published six years ago.The mechanism controlling the development of this unique histological formation is still unknown.Further,the prognosis of patients with OGC involvement is also controversial.CASE SUMMARY We report the case of a 48-year-old woman,who presented to the outpatient department with a palpable,growing,painless mass in her left breast for about one year.Sonography and mammography revealed a 26.5 mm ×18.8 mm asymmetric,lobular mass with circumscribed margin and the Breast Imaging Reporting and Data System was category 4C.Sono-guided aspiration biopsy revealed invasive ductal carcinoma.The patient underwent breast conserving surgery and was diagnosed with invasive breast carcinoma with OGCs,grade Ⅱ,with intermediate grade of ductal carcinoma in situ(ER:80%,3+,PR:80%,3+,HER-2:negative,Ki 67:30%).Adjuvant chemotherapy and post-operation radiotherapy were initiated thereafter.CONCLUSION As a rare morphology of breast cancer,breast carcinoma with OGC occurs most often in relatively young women,has less lymph node involvement,and its occurrence is not racedependent.

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.

HISTOCHEMICAL CHANGES OF HOST CELLS IN TISSUE CULTURE INOCULATED WITH RICKETTSIA TSUTSUGAMUSHI

Investigation of metabolic changes of host cells in tissue culture caused by Rickettsia tsutsugamushi helps to elucidate the pathogenesis and mechanism of immunity in scrub typhus. We used the histochemical methods for observing the changes of nucleic acids in the rabbit testicular monolayer tissue culture inoculated with R. tsutsugamushi. The method of preparation and inoculation of the tissue culture was the same as already reported. In the present experiment a glass slide was first inserted into the tube. before introduction of tissue cells. Examination was made once every 4 days after the cells had been inoculated with rickettsia. On the days

肿瘤标记物的分类、筛选及其在肿瘤病理诊断中的应用

随着免疫组织化学研究的进展,各种新的肿瘤标记物的发现对肿瘤的来源和罕见肿瘤的诊断、以及转移瘤原发部位的确定,提供了客观而可靠的依据。但是免疫组化还面临不少问题。越来越多的肿瘤标记物的相对特异性和交叉反应,未分化或低分化肿瘤不表达有关抗原,给诊断带来困难。尤其是软组织肿瘤,由于组织起源的多样性和多向分化的潜能。

Rice Repetitive DNA Sequence RRD3:a Plant Promoter and Its Application to RNA Interference

Previously, a moderately repetitive DNA sequence （RRD3） was cloned from rice （Oryza sativa L.） by DNA renaturation kinetics. Sequence analysis revealed several conserved promoter motifs, including four TATA-boxes and a CAAT-box, and promoter activity was shown in Escherichia coli and mammalian expression systems. Here, we inserted the RRD3 fragment into the plant promoter-capture vector, pCAMBIA1391Z, and examined whether the RRD3 fragment has promoter activity in plants. Transgenic tobacco and rice calli both showed β-glucuronidase （GUS） activity, indicating that RRD3 can act as a promoter in both monocot and dicot plants. Based on the promoter characteristic of RRD3, we designed a plant universal binary vector, pCRiRRD3, which is suitable for performing researches on plant RNA interference. This vector has two multiple cloning sites to facilitate sense and antisense cloning of the target sequence, separated by an intron fragment of 200 bp. The efficiency of the vector for gene silencing was assayed by histochemical and quantitative fluorometric GUS assays in transgenic tobacco. These research results suggested that this plant RNAi vector pCRiRRD3 can effectively perform gene silencing researches on both monocot and dicot plants.

Improved germination of threatened medicinal Prunus africana for better domestication: effects of temperature, growth regulators and salts

Prunus africana is an endangered medicinal species and has been classified as a priority for domestication in Cameroon.However,the seeds rapidly lose their viability during storage at room temperature after 2–3 months.This study aimed to improve seed germination of P.africana by germinating at different temperatures(4℃,28℃)using different concentrations of growth stimulators(sodium nitrate and gibberellic acid)and different concentrations of salts.P.africana seeds had 91.7%germination at 4C after 1 month of storage.Growth regulators considerably influenced germination after 6 months and reached 66.0%with 10 mM gibberellic acid and 100%with 10 mM sodium nitrate.Approximately three shoots per seed were developed,an indication of polyembryony.Histochemical analyses revealed the presence of protein-like bodies close to the embryo axis and accumulations of starch after 7 days of germination.After 14 days,amyloplasts and dark protein bodies of various sizes were observed.The outcome of this study will contribute to improve the germination of P.africana for better domestication and conservation.

Detection of Ca^(2+)-dependent acid phosphatase activity identifies neuronal integrity in damaged rat central nervous system after application of bacterial melanin

The study aims to confirm the neuroregenerative effects of bacterial melanin （BM） on central nervous system injury using a special staining method based on the detection of Ca^2＋-dependent acid phosphatase activity. Twenty-four rats were randomly assigned to undergo either unilateral destruction of sensorimotor cortex （group I; n = 12） or unilateral rubrospinal tract transection at the cervical level （C3-4） （group II; n = 12）. In each group, six rats were randomly selected after surgery to undergo intramuscular injection of BM solution （BM subgroup） and the remaining six rats were intramuscularly in）ected with saline （saline subgroup）. Neurological testing confirmed that BM accelerated the recovery of motor function in rats from both BM and saline subgroups. Two months after surgery, Ca^2＋-dependent acid phosphatase activity detection in combination with Chilingarian＇s calcium adenoside triphosphate method revealed that BM stimulated the sprouting of fibers and dilated the capillaries in the brain and spinal cord. These results suggest that BM can promote the recovery of motor function of rats with central nervous system injury; and detection of Ca^2＋-dependent acid phosphatase activity is a fast and easy method used to study the regeneration-promoting effects of BM on the injured central nervous system.

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.

Cloning and functional analysis of the promoter of allergen gene Ara h 1 from peanut

Peanut seeds are ideal bioreactors for the production of foreign recombinant proteins or secondary metabolites.Seed-specific promoters(SSPs)can direct the expression of genes specifically in seeds to avoid undesirable effects associated with constitutive expression.However,few SSPs have been identified in peanut.Previous studies have shown that some allergen-encoding genes encode seed storage proteins or exhibit seed-specific/preferential expression.In this study,we characterized allergen-encoding genes from across the genomes of Arachis species to explore seed-specific genes.We found that at least 9 out of 16 identified peanut allergen-encoding genes were expressed specifically in the seeds or were preferentially expressed.A 1493-bp promoter fragment of allergen gene Ara h 1(we named it AHSSP6)was isolated from cultivated peanut genome.cis-element analysis showed that three RY repeat elements which usually exsisted in seed or embryo specific promoter sequence were also present in AHSSP6 sequence.Histochemical analysis showed AHSSP6 could drive the expression of aβ-glucuronidase(GUS)reporter gene specifically in the seeds or cotyledon tissue of transgenic Arabidopsis,while not in other tissues.These findings indicated that these promoters of allergen genes were candidate SSPs,and AHSSP6 was a novel SSP which could be potentially utilized in peanut improvement.