Intestinal metaplasia,dysplasia and gastric cancer:a study of mucohistochemistry,immunohistochemistry and cell DNA quantitative analysis

One hundred and sixty-eight specimens of intestinal metaplasia(IM)with variousaccompanying lesions in gastric mucosa were studied with mucohistochemical and ABCimmunohistochemical staining,The quantitative analysis of cell DNA was done withflowcytometry for 36 specimens.The results indicated that the incidence of type Ⅱb IM wassignificantly higher in the groups of dysplasia(34.6％)and mucosa adjacent to gastric cancer(GC)(51.7％)than in the chronic atrophic gastritis(CAG)group(16.0％)(P【0.01).The expres-sion rate of monoclonal antibody MG7 related antigen(MG7-Ag)in type Ⅱb IM(473％)wasalso significantly higher than those in type la(29.7％),Ib(26.1％)and Ⅱa IM(28.3％)(P【0.05).Expression rate of MG7-Ag,DNA aneuploid rate and percentage of S phase cell werestatistically higher in the type Ⅱb IM with dysphsia(62.5％,62.5％ and 143±32)than in typeⅡb 1M without dysplasia(47.3％,12.5％and12.7±2.9)(P【0.05 and P【0.01).These findingssuppor the supposed progressive process:CAG→type Ⅱb IM→dysplasia→GC,andtype Ⅱb IM with dysplasia is closely associated with GC.

High-efficiency somatic embryogenesis and morphohistology and histochemistry of somatic embryo development in Larix leptolepis Gordon

A high-efficiency somatic embryogenesis protocol of Japanese larch （Larix leptolepis Gordon） has been established in our investigation. Calli were induced from immature zygotic embryos of female cones ofL. leptolepis and then subcultured regularly on to a modified Gupta and Durzan （DCR） basal medium for 5 years. Embryogenic tissues showed distinct morphological changes dur- ing somatic embryo development when they were transferred to a maturation medium supplemented with abscisic acid （ABA） com- pared with the morphology in a medium lacking ABA. Histological observations indicated that polyembryony was a characteristic feature during early embryogeny and somatic embryos at later stages showed normal histodifferentiation. In addition, histochemical analysis revealed that abundant starch granules and proteins accumulated in mature embryos, indicating that they played important roles in the development and regeneration of normal plantlets from somatic embryos on hormone-free germination media

Histochemistry of microinfarcts in the mouse brain after injection of fluorescent microspheres into the common carotid artery

The mouse model of multiple cerebral infarctions,established by injecting fluorescent microspheres into the common carotid artery,is a recent development in animal models of cerebral ischemia.To investigate its effectiveness,mouse models of cerebral infarction were created by injecting fluorescent microspheres,45–53μm in diameter,into the common carotid artery.Six hours after modeling,fluorescent microspheres were observed directly through a fluorescence stereomicroscope,both on the brain surface and in brain sections.Changes in blood vessels,neurons and glial cells associated with microinfarcts were examined using fluorescence histochemistry and immunohistochemistry.The microspheres were distributed mainly in the cerebral cortex,striatum and hippocampus ipsilateral to the side of injection.Microinfarcts were found in the brain regions where the fluorescent microspheres were present.Here the lodged microspheres induced vascular and neuronal injury and the activation of astroglia and microglia.These histopathological changes indicate that this animal model of multiple cerebral infarctions effectively simulates the changes of various cell types observed in multifocal microinfarcts.This model is an effective,additional tool to study the pathogenesis of ischemic stroke and could be used to evaluate therapeutic interventions.This study was approved by the Animal Ethics Committee of the Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences(approval No.D2021-03-16-1)on March 16,2021.

Effects of Natural Radiation, PAR and Artificial Ultraviolet Radiation-B on the Ultrastructure and Histochemistry of Leaf of Oryza sativa L.

Ultraviolet radiation-B (UVBR) affects plants in many important ways, including reduced growth rate, reduction of primary productivity and changes in ultrastructure. The rice (Oryza sativa) is one of the most cultivated cereal in the world along with corn and wheat, representing over 50% of agricultural production. In this study, we examined O. sativa exposed to natural radiation denominated which “ambient samples”, plants cultivated which photosynthetically active radiation (PAR), denominated with PAR-only and plants cultivated with PAR + UVBR for 2 h per day during 30 days of cultivation in vitro. The samples were processed for electron microscopy and histochemistry analysis. PAR + UVBR caused changes in the ultrastructure of leaf of O. sativa, mesophyll cells, which included increased thickness of the cell wall and plastoglobuli, reduced intracellular spaces, changes in the cell contour, and destruction of chloroplast and mitochondria internal organization. The exposure to PAR + UVBR led to changes in guard and subsidiary cells, and the stomata and papillae were with irregular shape. The reduction of epicuticular wax that covered the leaf, was observed. Taken together, these ?ndings strongly suggested that PAR + UVBR negatively affects the ultrastructure and morphology and growth rates, of leaf of O. sativa and, in the long term, their economic viability.

A Study of Histology and Enzymatic Histochemistry on Rabbit's Retina in Acute Ocular Hypertension

The changes of activities of enzymes relating to energy metabolism in rabbit's retina in acute ocular hypertension were observed. The activities of succinate dehydrogenase and adenosine triphosphatase were found to be reduced, while the activities of the lactatic dehydrognease and glucose-6-phosphatase increased. The results revealed the metabolic disturbance of energy in retina after acute ocular hypertension might be the underlying factors relating to the defects of the functions and structures of the...

Enzymatic Histochemistry of Retina with Experimental Intraocular Pressure Elevation in Rabbits

To investigate the pathogenesis of retina lesions caused by intraocular pressure elevation, activities and distribution of enzymes in retina including lactic dehydrogenase (LDH), succinate dehydrogenase (SDH), adenosinetriphosphatase (ATPase), acid phosphatase (ACP), cholinesterase (ChE), cytochrome oxidase(CCO),nucleotidase (5'-Nase) and glucose-6-phosphatase (G6Pase) were determined histochemically in 30 rabbits. It was found that 1) in the early stage of intraocular pressure elevation, the activities of LDH, SDH, ATPase, ACP, and ChE in retina were increased, while the activities of CCO,5'-Nase decreased;2)in thelate stage of intraocular pressure elevation, the activities of all these enzymes but ACP, which showed a reduced activity, were close to the normal level; 3) in superoxide dismutase.(SOD-CCE) treated group, except the slight increase of LDH and G6Pase activities,the activities of the remaining enzymes were near to normal. Our results suggest that the various histochemical changes in retina induced by intraocular pressure elevation were cornpensatory in the early stage and were beneficial to the supply of energy needed in retinal tissue andcellular metabolism;while in the late stage, the lesion of retina cells developed due to decompensation.SOD-CCE could alleviate the retinal lesions caused by intraocular pressure elevation, and can be used as auxiliary drug for the treatment of intraocular pressure elevation.

The 12th China-Japan Joint Seminar on Histochemistry and Cytochemistry

01 Differentiation-dependent changes in DNA methylation and histone H3 acetylation: possible roles of these epigenetic factors in mouse spermatogenesis Takehiko Koji Department of Histology and Cell Biology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8523, Japan Mammalian spermatogenesis is a well-ordered process, consisting of spermatogonial mitosis, spermatocytic meiosis and spermiogenesis.The process is also characterized by successive chromatin condensation and a high frequency of spermatogenic cell apoptosis.Therefore, epigenetic factors can be implicated in the regulation of spermatogenesis and any epigenetic disorder may lead to the induction of apoptosis. When we analyzed global level of DNA methylation by immunohistochemistry with anti-5-methylcytosine in paraffin-embedded sections of mouse testis, there was no difference among any stages of germ cells.

ACETYLCHOLINESTERASE HISTOCHEMISTRY OF THE THALAMUS IN THE PRIMATE

Objective To observe the distribution of acetylcholinesterase activity in the thalamus of the monkey.Methods Histochemical method was used to detect the acetylcholinesterase activity in the thalamus.Results Acetylcholinesterase was found to be inhomogeneous distribution in the primate thalamus and to reveal previously uncovered inhomogeneity within certain thalamic nuclei and their subdivisions. The medial, ventral and posterior nuclear groups displayed markedly uneven acetylcholinesterase reaction.In the mediodorsal nucleus,three distinct sbudivisions were revealed by acetylcholinesterase histochemistry, medial magnocellular part, ventral sector of central parvicellular part and dorsolateral sector of lateral pars multiformity showed weak, moderate and strong acetylcholinesterase activity, respectively. In the ventral nuclear group, acetylcholinesterase histochemistry was strong in the medial part of ventral posterior nucleus, moderate in the magnocellular part of ventral anterior, caudal, medial, oral and pars postrema parts of ventral lateral nucleus, as well as lateral part of ventral posterior nucleus, poor and weak in the inferior part of ventral posterior nucleus, par compacta of the medial part of ventral posterior nucleus and parvicellular part of ventral anterior nucleus. In the pulvinar nucleus, acetylcholinesterase reaction ranged from weak, moderate to strong in the parts of the oral, medial and lateral, as well as inferior of this nucleus, respectively. Regional variations of acetylcholinesterase activity within the thalamic nuclei and their subdivisions can help to identify them by acetylcholinesterase histochemistry. In addition, the dark patches of strong acetylcholinesterase activity contrasting with a lighter surrounding matrix were revealed within the parvicellular part and pars multiformis of mediodorsal nucleus, paracentral nucleus, central lateral nucleus, pars postrema part of ventral lateral nucleus and medial habenula nucleus, as well as medial part of pulvinar necleus, respectively.Conclusion Acetylcholinesterase histochemistry can serve as useful marker for delineating the thalamic nuclei and their sbudivisions. The different distribution of acetylcholinesterase histochemistry implies the activity of thalamic nuclei may be modulated by cholinergic systems.

STUDIES OF NUCLEOLAR ORGANIZER REGIONS AND MUCIN HISTOCHEMISTRY IN ULCERATIVE COLITIS

The argyrophil method for nucleolar organizer regions (AgNOR) known as the AgNor technique and mucin histochemical stain were applied to Investigate the dysplasia and cancaration in ulcerative colitis Including 58 cases of biopsy specimens and three cases operative specimens. The numbers of AgNOR have been gradully increased with the grades of dasplasls. Similarly the percentage of sialomucin having vicinal diols on side chain of sialic acid also gradully Increased with the grades of dysplasia. The AgNOR reflects rDNA transcriptional activity responsible for degrees of differentiation of cell. Epithelial cells secreting a heterogeneous mucin, could be taken as a signal of abnormal cellular differentiation. AgNOR and mucln chages might be assumed as a criteria In representing malignant transformation.

Myelin histology:a key tool in nervous system research

The myelin sheath is a lipoprotein-rich,multilayered structure capable of increasing conduction velocity in central and peripheral myelinated nerve fibers.Due to the complex structure and composition of myelin,various histological techniques have been developed over the centuries to evaluate myelin under normal,pathological or experimental conditions.Today,methods to assess myelin integrity or content are key tools in both clinical diagnosis and neuroscience research.In this review,we provide an updated summary of the composition and structure of the myelin sheath and discuss some histological procedures,from tissue fixation and processing techniques to the most used and practical myelin histological staining methods.Considering the lipoprotein nature of myelin,the main features and technical details of the different available methods that can be used to evaluate the lipid or protein components of myelin are described,as well as the precise ultrastructural techniques.

A study of histochemistry on response of ventricular myocardium to radiofrequency catheter ablation

Abstract Using histochemistry technique, the effects of radiofrequency catheter ablation (RFCA) on the activities of LDH, SDH, CCO, and Ca^2+ ATPase of guinea pig ventricular myocytes were examined. In the meanwhile, the histological changes were observed as Department of Cardiology, Tongji Hospital, Tongji Medical University. Wuhan 430030, China (Wang YG, Lu ZY) Department of Anatomy, Tongji Medical University, Wuhan 430030, China (Xiong XK) control. Radiofreqency energy (500 kHz) delivered was 20W×10s. The resultes were as follows: RFCA resulted in significant impairments in all the four kinds of enzymses but without statistical differences among their areas involved in this energy level, and there are no statistical significant differences when compared with those of histological lesion area. These findings showed consistency in areas of the histological and histochemical lesions resulted from RFCA.

Histological assessment in peripheral nerve tissue engineering

The histological analysis of peripheral nerve regeneration is one of the most used methods to demonstrate the success of the regeneration through nerve conduits. Nowadays, it is possible to evaluate different parameters of nerve regeneration by using histological, histochemical, immunohistochemical and ultrastructural techniques. The histochemical methods are very sensible and are useful tools to evaluate the extracellular matrix remodeling and the myelin sheath, but they are poorly specific. In contrast, the immunohistochemical methods are highly specific and are frequently used for the identification of the regenerated axons, Sehwann cells and proteins associated to nerve regeneration or neural linage. The ultrastructural techniques offer the possibility to perform a high resolution morphological and quantitative analysis of the nerve regeneration. However, the use of a single histological method may not be enough to assess the degree of regeneration, and the combination of different histological techniques could be necessary.

CLINICOPATHOLOGIC FEATURES AND DIAGNOSISOF COMBINED HEPATOCELLULAR ANDCHOLANGIOCARCINOMA

Hepatoma cases(N=130) were analyzed through histochemical and immunohistochemical staining. There were 99 cases of hepatocellular carcinoma(HCC), 15 cholangiocarcinoma(CC), and 16 combined HCC and CC (HCC+CC). The clinical features and the cases accompanied with hepatitis and/or liver cirrhosis in the non-tumor liver tissue of HCC+CC Group were between HCC GrouP and CC Group. Histologically, there were 4 cases with trabecular, 4 with pseudoglandula, 3 with solid type in HCCWC Group. In these 11 cases, the CC area was less than 10% of the neoplasm- The cases were classified as HCCWC type I. There was no obvious stroma fibrosis. The rest 5 cases of HCC+CC showed tubular carcinoma. The CC area took up over 10% of the tumor. These cases were designed as HCC+CC type II. There was significant fibrosis in the stroma so that its CC area is indistinguishable from that of CC cases. The CC area of all HCC+CC cases was positive to mucin and EMA staining, the same to that of CC cases. Near 70% of the HCC+CC cases had intracytoplasmic glycogen in the HCC area. The HCC area was mucin and EMA negative, similar to that of HCC cases- We also observed transition areas between HCC and CC in both of the type I and type II HCC+CC cases. The areas were mucin negative but EMA positive. We concluded that HCC+CC bad HCC and CC area. with the characteristics of HCC of CC respectively. Histochemical mucin staining and immunohistochemical EMA staining were valuable in detection and diagnosis of the HCC+CC.

Stain Materials’Role in Biological Research:A Tool in Heath Care

Stains and staining methods significantly assist in diagnoses in medical research and health care.Certain color of a dye can identify the location of a tumor within a specimen.Applying histochemical-staining enabled morphological identification of fibrin in the lymphoid tissue during cancer progression.Staining methods in combination with the LSFM(light-sheet fluorescence microscopy)allowed tracing the drug penetration,development and spread of tumors.The Curcumin dye is in use for labelling and imaging of Aβplaques in post-mortem brain tissue.Immunofluorescent staining methods are employed in detection of some important proteins in early diagnostic changes relevant to heart damage.The methods are developed in medical research to include stem cells and tissue engineering,cell culturesproperties and capabilities,connective tissues and extracellular matrix,nervous system,musculoskeletal system;respiratory system,liver and gastrointestinal tract,and male and female reproductive systems.

Effects of AIBL on Oncomelania hupensis,the intermediate snail host of Schistosoma japonicum:An enzyme histochemical study

Objective:To explore the effect of AIBL on Oncomelania hupensis,the intermediate snail host of Schistosoma japonicum.Methods:The enzyme histochemical profiles of cholineslerase, cytochrome oxidase,lactate dehydrogenase,nitric oxide synthase,and succinate dehydrogenase in the soft tissues of Oncomelania hupensis,the intermediate host snail of Schistosoma japonicum, were analyzed before and after treatment with the active ingredient of Buddleia lindleyana(AIBL), a potent and safe plant molluscicide.Results:Treatment with AIBL induced a notable decrease in the activities of the five enzymes(P【0.01).Conclusions:The results indicate that AIBL impairs the activities of the enzymes,thereby influencing the transfer of neurotransmitter and energy supply in Oncomelania hupensis and ultimately harming their various physiological functions, which are considered to cause death of the species.

Arylsulfatase,β-galactosidase and lysozyme in gastric cancer cells and its relationship to invasion

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Cytomegalovirus Infection in Children's Liver Tissues

Immunoperoxidase histochemical assay with monoclonal antibody against human cytomaglovirus (HCMV) was used to detect immediate early antigen (IEA) and early antigen (EA) of HCMV infection in liver tissue of 72 podiatric cases (34 autopsies and 38 biopsies). The HCMV antigen was positive in 25 % (18/27). Among them, 12 cases were both HCMV-IEA and EA positive; 4 were HCMV-IEA positive and 2 HCMV-EA positive only. Liver HCMV infection rate in neonates, the infants with the age <1 year and >l year was 8. 0 %,60. 0 %, and 14. 8 %, respectively, indicating that liver HCMV infectioh occurred at various ages. The liver HCMV infection rate in different diseases was 50. 0 % in infantile hepatitis syndrome; 70. 0 % in extrabiliary malformation, and 12. 5 % in other hepatopathies, suggesting that infantile hepatitis syndrome and extrabiliary malformation were related with HCMV infection in liver tissues.

Black soldier fly and gut health in broiler chickens:insights into the relationship between cecal microbiota and intestinal mucin composition

Background:The relationship between diet and intestinal microbiota and mucin composition appears to be fundamental for poultry gut health.The effects of insect meal(whose role as alternative feed ingredient is now well recognized)on gut microbiota and mucin composition have recently been reported in Tenebrio molitor-fed free-range and broiler chickens,but no data are currently available for Hermetia illucens(HI)-fed broilers.The present study evaluated the effects of dietary HI meal inclusion on cecal microbiota and intestinal mucin composition of broiler chickens.Results:A total of 256 male broiler chickens were allotted to 4 dietary treatments(control diet[C]and 5%,10%and 15%HI meal inclusion,with 8 replicate pens/treatment and 8 birds/pen)and slaughtered at 35 d of age(2 animals/pen,16 birds/diet).The cecal microbiota assessment by 16S rRNA amplicon based sequencing showed lower alpha diversity in HI15 chickens(Shannon,P<0.05)and higher beta diversity(Adonis and ANOSIM,P<0.001)in birds fed HI diets than C.Furthermore,HI15 birds displayed significant increase of the relative abundance of Proteobacteria phylum(False Discovery Rate[FDR]<0.05)when compared to HI10.L-Ruminococcus(Ruminococcus from Lachnospiraceae family),Faecalibacterium,Blautia and Clostridium genera were found to be characteristic of HI5 cecal microbiota(FDR<0.05),while broiler chickens fed HI10 and HI15 diets were characterized(FDR<0.05)by Lactobacillus and Ruminococcus(HI10)and Bacteroides,Roseburia and Helicobacter genera(HI15).Periodic-acid Schiff,Alcian Blue pH 2.5 and high iron diamine staining on small and large intestine also demonstrated lower mucin staining intensity in the intestinal villi of HI10 and HI15 birds than C(P<0.05).Conclusions:Dietary HI meal utilization at low inclusion levels(i.e.,5%)positively influenced either the cecal microbiota or the gut mucin dynamics in terms of selection of potentially beneficial bacteria and increase in villi mucins.However,high inclusion levels(in particular the 15%)may have a negative influence in terms of partial reduction of microbial complexity,reduction of potentially beneficial bacteria,selection of bacteria with mucolytic activity and decrease in villi mucins.

Effect of Hypertension on Hearing Function,LDH and ChE of the Cochlea in Older Rats

The relationship between the hypertension and the aging process of hearing organ was investigated Twenty Wistar 3-month old rats and 20 Wistar 12-month old rats, 20 spontaneously hypertensive rat stroke-prone (SHRSP) 3-month old rats and 20 SHRSP 12-month old rats free of middle ear infections as observed under otomicroscopy, with normal tympanic membrane and auricle reflex, were selected to be divided into two experimental groups and two control groups respectively The tail artery blood pressure was measured non-invasively The threshold of auditory brainstem response (ABR) was measured by Spirit TM evoked potential meter The LDH and ChE staining in the inner ear was performed and the optical density was analyzed by the HPIAS analysis system The results showed that there was no difference in the ABR thresholds, the activities of LDH and ChE between Wistar 3-month old group and SHRSP 3-month old group ( P >0 05) The mean value of ABR threshold and the activities of LDH and ChE in the Wistar 12-month old group at relevant sections were significantly greater than those in the two 3-month old groups ( P< 0 05), whereas the mean value of ABR threshold and the activities of LDH and ChE in the SHRSP 12-month old group at relevant sections were significantly higher than those in the 3-month old control group ( P< 0 01) It was concluded that presbycusis existed in the Wistar 12-month old group rats The glycogenosis and the abnormal secretion of neural transmitter were discerned after hypertension All the above factors may worsen the aging of the hearing system

Anatomical, Histochemical and Cytogenetic Features of <i>Doryopteris triphylla</i>(Pteridaceae)

Doryopteris triphylla (Pteridaceae-Cheilanthoideae) grows in xeric habitats in Brazil, Paraguay, Uruguay and Argentina. The aim of this study was to characterize D. triphylla anatomically, histochemically and cytogenetically. For anatomical characterization, rhizomes, roots, petioles and leaves were made and then stained using Safranine-Astra Blue for further observations. Leaf blades were also cleared. For histochemical analysis, leaf cross sections were stained with different reagents to identify glandular trichomes compounds. For cytogenetic characterization, a karyogram was performed using laboratory cultivated roots. Results show a dictyostelic rhizome covered with scales with apical secreting gland;diarch roots;petiole cross-sections show thick cuticle, uniseriate epidermis, parenchymatic cortex cells with thick walls and a vascular bundle with two xylem groups;and hypostomatic fronds with glandular trichomes. Histochemical studies of secretion products of the glandular trichomes were positive for polysaccharides, pectins, lipids, acid lipids, dihydroxyphenols, phenols and flavonoids. Cytogenetically, D. triphylla is described as a diploid species (2n = 60), with chromosomes gradually decreasing in size. The apical glands in scales of rhizomes, the presence of two xylem groups in the vascular bundle in the petiole and the glandular trichomes on the abaxial surface are new contributions to the species. The type of chemical products secreted by glandular leaf trichomes and karyotype estimation is shown for the first time in this species.