Effects of propentofylline on CNS remyelination in the rat brainstem

Propentofylline (PPF) is a xanthine derivative with pharmacological effects distinct from those of the classical methylxanthines. It depresses activation of microglial cells and astrocytes which is associated with neuronal damage during neural inflammation and hypoxia. The aim of this study was to evaluate whether PPF had the capacity of affecting glial cells behavior during the process of demyelination and remyelination following ethidium bromide (EB) gliotoxic injury. EB injection into the CNS is commonly used as an experimental demyelinating model inducing local oligodendroglial and astrocytic death, which results in primary demyelination, blood–brain barrier and glia limitans disruption and Schwann cells invasion. Sixty Wistar rats were divided into four different groups receiving 10 microlitres of 0.1% EB or 0.9% saline solution into the cisterna pontis and treated or not with the xanthine. PPF treatment was done using 12.5 mg/kg/day by the intraperitonial route for 31 days of the experimental period. The rats were euthanized from 7 to 31 days after EB injection and brainstem sections were collected and processed for light and transmission electron microscopy studies. Results from both groups were compared by using a semi‐quantitative method developed for documenting in semithin sections the extent and nature of remyelination of demyelinating lesions. Results showed that PPF administration after EB injection significantly increased both oligodendroglial and Schwann cell remyelination at 31 days (mean remyelination scores of 3.67 ± 0.5 for oligodendrocytes and 1.27 ± 0.49 for Schwann cells) compared to untreated animals (scores of 3.19 ± 0.57 and 0.90 ± 0.33, respectively). Microsc. Res. Tech. 77:23–30, 2014. © 2013 Wiley Periodicals, Inc.     

Measuring microenvironment mechanical stress of rat liver during diethylnitrosamine induced hepatocarcinogenesis by atomic force microscope

We developed a highly sensitive method to detect liver tissue stiffness with atomic force microscopy (AFM), and investigated the physical features of hepatocarcinogenesis. Wistar rats received weekly intraperitoneal injections of diethylnitrosamine (DEN) or saline (control) followed by a 2-week wash-out period. Liver samples were harvested at 10, 14, or 18 weeks for pathological examination and stress detection. Previously normal liver tissues developed fibrosis and carcinoma after DEN administration. Although the elastic modulus (E) values of the normal (saline; 0.18 ± 0.04 MPa), fibrotic (8 weeks DEN; 0.25 ± 0.06 MPa) and cirrhotic (12 weeks DEN; 0.39 ± 0.06 MPa) tissues were significantly different, there was no significant difference between the E values of the cirrhotic and the hepatic cell carcinoma (16 weeks DEN; 0.42 ± 0.07 MPa) tissues. Thus, tissue stiffness quantitatively increases during hepatocarcinogenesis, and AFM can be used to sensitively and precisely detect liver stiffness at the microscopic level. Microsc. Res. Tech. 2009. © 2009 Wiley-Liss, Inc.     

Astaxanthin delayed the pathogenesis of diabetic nephropathy in type 1 diabetic rats

This study was designed to investigate the protective effects of Astaxanthin (AST) in rats with diabetes mellitus (DM) induced by streptozotocin. SD rats were divided into control group (n = 5, only received normal saline), DM group (n = 8) and AST + DM group (n = 8; AST: 50 mg/kg/day). DM rats were induced by intraperitoneal injection of streptozocin (STZ, 65 mg/kg). Blood glucose level and body weight were determined at weeks 0, 2, 4, 6 and 8, respectively. At week 8, kidney function was determined, together with expression of P53 and dynamin-related protein-1 (Drp1) by Western blot analysis and immunofluorescence. AST led to increase of body weight in rats with DM. AST + DM group showed a significant decrease in blood glucose level at week 4 compared with DM group (P < 0.05). AST improved renal function and significantly reduced expression of P53 and Drp1 in DM rats. In addition, AST can effectively reduce the blood glucose in DM rats, and delayed the pathogenesis of diabetic nephropathy. Such delay mediated by AST may be associated with the downregulation of Drp1 and P53.     

Protective effect of aqueous suspension of dried latex of <i>Calotropis procera</i> against oxidative stress and renal damage in diabetic rats

Calotropis species have been used in the traditional medicinal system for the treatment of diseases of the liver and abdomen. In view of the antioxidant and anti-hyperglycemic properties of an aqueous suspension obtained from the dried latex of Calotropis procera, the present study was carried out to evaluate its efficacy in affording protection against alloxan induced changes in rat kidney. A single intraperitoneal injection of alloxan (150 mg/kg) in rats produced hyperglycemia within 3 days and altered kidney functions over a period of 90 days. Daily oral administration of the aqueous suspension (100 and 400 mg/kg) in diabetic rats produced anti-hyperglycemic effect that was comparable to that of glibenclamide (10 mg/kg). Unlike glibenclamide, the aqueous suspension did not increase the serum insulin levels in diabetic rats. However, it produced a marked reduction in the levels of urinary glucose and protein and normalized the renal tissue levels of thiobarbituric acid-reactive substances (TBARS) and glutathione (GSH) in diabetic rats and the effect was comparable to that of glibenclamide. The protection afforded by the aqueous suspension was also evident from the histological analysis of the renal tissue. Our study shows that by exhibiting antioxidant and anti-hyperglycemic property the aqueous suspension of dried latex of C. procera affords protection against the complications associated with diabetes.     

Protective effect of aqueous suspension of dried latex of Calotropis procera against oxidative stress and renal damage in diabetic rats

Calotropis species have been used in the traditional medicinal system for the treatment of diseases of the liver and abdomen. In view of the antioxidant and anti-hyperglycemic properties of an aqueous suspension obtained from the dried latex of Calotropis procera, the present study was carried out to evaluate its efficacy in affording protection against alloxan induced changes in rat kidney. A single intraperitoneal injection of alloxan (150 mg/kg) in rats produced hyperglycemia within 3 days and altered kidney functions over a period of 90 days. Daily oral administration of the aqueous suspension (100 and 400 mg/kg) in diabetic rats produced anti-hyperglycemic effect that was comparable to that of glibenclamide (10 mg/kg). Unlike glibenclamide, the aqueous suspension did not increase the serum insulin levels in diabetic rats. However, it produced a marked reduction in the levels of urinary glucose and protein and normalized the renal tissue levels of thiobarbituric acid-reactive substances (TBARS) and glutathione (GSH) in diabetic rats and the effect was comparable to that of glibenclamide. The protection afforded by the aqueous suspension was also evident from the histological analysis of the renal tissue. Our study shows that by exhibiting antioxidant and anti-hyperglycemic property the aqueous suspension of dried latex of C. procera affords protection against the complications associated with diabetes.     

The changes of gap junctions between pituitary folliculo‐stellate cells during the postnatal development of zucker fatty and lean rats

We investigated the effect of leptin on the postnatal development of gap junctions between folliculo‐stellate cells by using Zucker fatty (fa/fa) rats that have defects of the functional leptin receptor. Male Zucker fatty rats (fa/fa) and male Zucker lean rats (+/+) were used at each of the following postnatal ages: 20, 30, 40, 50, 60, 70, 80, 90 days, and 1 year. On one of the aforementioned dates, the anterior pituitary glands were prepared for observation by transmission electron microscopy. We quantified the number of follicles and gap junctions, and calculated the rate of occurrence as the ratio of the number of gap junctions existing between folliculo‐stellate cells per intersected follicular profile. In Zucker lean male rats, the number of gap junctions remained relatively constant from days 50 to 90 (0.44 ± 0.02 to 0.49 ± 0.03), and was similar in 1 year old rats (0.47 ± 0.03). These data were statistically higher compared to Zucker fatty male rats. In Zucker fatty male rats, very few gap junctions were observed in 30‐day‐old rats (0.04 ± 0.01: mean ± SE). This disruption of gap junction formation persisted, and the number of gap junctions remained constant and showed a low level from days 40 to 90 (0.11 ± 0.02 to 0.17 ± 0.02); this finding was similar in 1‐year‐old rats (0.17 ± 0.02). These observations indicate that the effect of leptin over the gap junction formation within the anterior pituitary glands was directly mediated by interaction with the functional leptin receptor present on the folliculo‐stellate cells. Microsc. Res. Tech. 77:31–36, 2014. © 2013 Wiley Periodicals, Inc.     

Observation of curative effect of intravitreal injection of conbercept in wet age‐related macular degeneration: Optical coherence tomography analysis after injection

To observe the clinical efficacy of intravitreal injection of conbercept in the treatment of wet age‐related macular degeneration (wAMD), optical coherence tomography (OCT) and the best corrected visual acuity (BCVA) was observed to measure the changes of anatomical changes of central macular thickness (CMT) and the area and volume of retinal pigment epithelium (RPE) uplift. Fifteen patients (15 eyes) with wet AMD were enrolled in this study. All patients underwent intravitreal injection of conbercept of 0.05 mL once. After 1 week, 1 month, and 3 months, OCT and BCVA were used to examine and to compare with the preoperative and postoperative central macular thickness and RPE uplift area. BCVA (median) increased respectively from 0.12 ± 0.13 to 0.21 ± 0.15 at 1 week, to 0.90 ± 0.25 at 1 month, to 0.38 ± 0.17 at 3 months (p < .001). The thickness of central macular decreased from 500 ± 25 μm to 256 ± 19 μm, 221 ± 29 μm, and 215 ± 14 μm, respectively. The normal physiological structure and stratification of the macular area were clear gradually. Conbercept treatment of wet AMD can significantly improve visual acuity, after 1 month up to the plateau, 3 months of continuous drug injection can make the vision maintained at a high stage, and macular retinal normal structural morphology recovery is good, the treatment has no obvious adverse reactions, and with good security.     

Vascular smooth muscle orientation in straight portions of human cerebral arteries

A technique of embedding, sectioning and analysis has been developed for studying the orientation and proportional composition of smooth muscle in the straight portions of human major cerebral arteries. Various distortions, which occurred during processing and sectioning, were measured quantitatively. Nerve fibres were implanted as a reference frame within the paraffin block containing an arterial bifurcation. The nuclei of smooth muscle have been treated as three-dimensional vectors of cellular orientation. The projected length in the plane of the section and the section thickness were used to define section pitch. To relate these vectors to the overall geometry of cerebral arteries they were transformed such that the resultant pitch would be the same as that observed if the sections were cut normal to the arterial longitudinal axis. Matrix transformations of nuclear vectors were of the expansion and Eulerian forms. The average pitch for the five sections from three straight portions was ?0.22° ± 2.36 (SD of five means) with a range between ?2.8° and 3.2°. Because this pitch is small it is possible to use 6.9 μm thick longitudinal and mid-plane sections of straight arteries to obtain estimates of proportional composition. Stereological point counting was used to determine that smooth muscle comprised 72.0% ± 4.76 (SD for ten segments) of the tunica media in cerebral arteries.     

Identification of fenestrated capillary segments in microvascular corrosion casts of the rat exocrine pancreas

The capillary bed of the rat exocrine pancreas was studied by scanning and transmission electron microscopy of corrosion casts and tissue sections. Two types of capillaries were distinguished in corrosion casts. First, there were straight capillaries of relatively constant width (mean diameter 4.79±0.87 μm), which were characterized by numerous circular constrictions on their surface. About 37% of the capillaries belonged to this type. Second, there were undulating capillaries which showed smooth surfaced eccentric dilatations defined by similar surface constrictions. The bulging areas measured 8.43 ± 1.33 μm, the constrictions next to the bulges figured for 6.45 ± 1.53 μm. About 63% of the capillaries belonged to the second type. Two types of capillaries were also identified in tissue sections. First, there were capillaries with continuous endothelial lining (26% of capillary profiles; mean diameter 5.48 ± 1.67 μm); 27% of their endothelial lining was provided with underlying pericytes. Second, there were capillaries with fenestrated endothelium (64% of capillary profiles; mean diameter 6.16 ± 1.75 μm); 12% of their endothelial lining was accompanied by pericytes. According to frequency and dimension of these two types of capillaries, we conclude that bulged and undulating capillary casts correspond to fenestrated capillaries and straight capillary casts of constant width correspond to nonfenestrated capillaries. The frequency of crests on the surface of capillary casts correlates with the different frequency of pericyte processes on fenestrated and nonfenestrated capillaries. It is concluded that pericyte processes beneath the endothelium hold resistance against luminal pressure. Bulging areas of capillary casts correlate with fenestrated areas of endothelial lining, that is, areas which are not reinforced by pericyte processes.     

Dimensions of active cytochrome c oxidase in reconstituted liposomes using a gold ball shadow width standard: a freeze-etch electron microscopy study

The preparation and characterization of a distribution of gold balls on a thin, flat carbon film is described. The relation of the platinum carbon shadow width distribution means to a gold ball size is reported. Freeze-etched cytochrome oxidase vesicles and gold ball calibration grids were simultaneously shadowed with platinum/carbon. The shadow width distribution of the cytochrome oxidase located in and spanning the membrane was measured. The membrane fracture face edge and cross-fractured bilayer membrane edge were also measured. Dimensions of the cytochrome oxidase were found to be 5·8 ± 0·3 nm in diameter parallel to the membrane and 8·2 ± 0·3 nm long across the membrane. The bilayer membrane dimensions were 3·0 ± 0·3 nm for the half bilayer and 5·8 ± 0·3 nm for the cross-fractured bilayer membrane edge thickness. The length of the cytochrome oxidase was observed to span the bilayer membrane. Previous X-ray diffraction measurements on similar hydrated liquid crystalline artificial membranes were found to be in good agreement with the freeze-etched results. Membrane widths from thin-sectioned cytochrome oxidase vesicles were measured and found to be 5·8–5·9 nm in non-post-stained sections. Post-staining with uranyl acetate and/or lead citrate was shown to increase this average thickness. The technique of freeze-etching electron microscopy in conjunction with the gold ball shadow width calibration experiment has been shown to provide accurate and precise measurements of membranes and a functional intramembrane protein in a hydrated non-crystalline sample.     

A study of digital image analysis on the acrylamide derivative monomers induced apoptosis in rat cerebrum

Nowadays, apoptosis is mostly evaluated visually in histological studies. By using the quantitative digital image analysis, this study aimed to investigate the effect of acrylamide‐based monomers (acrylamide [AAm], methacrylamide [MAAm], N‐isopropylacrylamide [NIPAm]) on the cerebrum tissues in rats, which are the most common water‐soluble monomers in the production of polymeric hydrogels used as biomaterials. The Wistar albino rats weighing ~220–240 g were divided into control and three test groups. The control group received 1 mL of saline, and the test groups received 1 mL of aqueous 50 mg/kg/day intramuscular injection of AAm, MAAm, and NIPAm, respectively. At the end of the experiments, brain tissues of all rats euthanized by intramuscular injection of sodium pentobarbital were removed. Terminal deoxynucleotide transferase dUTP nick and labeling (TUNEL) method was applied to brain tissue sections. The monomers have been shown to cause apoptosis due to oxidative stress in cerebrum tissue. Based on apoptosis by tunneling method, quantitative digital image analysis of cell fragments was performed with Olympus cellSens Dimension 1.15 software, and the number, total count area, selected area, average area, and ROI% values of the fragments were found. In addition, the total area and ROI% values of the fragments increased linearly with increasing the molar mass of monomers from the digital image analysis data. Quantitative digital image analysis can facilitate the monitoring of apoptosis caused by the oxidative stress of monomers used in the production of the biomaterials.     

Ultrastructural study of the intracellular behavior of four mineral elements in the lactating mammary gland cells: study using conventional transmission electron microscopy

The effects of parenteral injection of aluminum, indium, gadolinium, or terbium in rats have been previously studied in several organs such as the liver, the kidneys, etc., but never in mammary glands. In this work, we have attempted to study the subcellular localization of these elements after their intraperitoneal administration. Their subsequent effects in the lactating mammary gland cells have also been studied. Our results using conventional transmission electron microscopy have shown that the lysosomes of the mammary glandular epithelial cells are the intracellular site of accumulation of the studied elements. Our results have also show intracellular deteriorations such as an expanded ergastoplasm and altered mitochondria after intraperitoneal injection of aluminum and indium.     

Second-order stereology and ultrastructural examination of the spatial arrangements of tissue compartments within glomeruli of normal and diabetic kidneys

The present study explores 3D spatial arrangements of compartments within the rat renal glomerulus and tests for differences after chemically-induced diabetes. In particular, the arrangements of capillaries, podocytes, mesangium and urinary space are quantified and compared between (a) kidneys within groups and (b) kidneys from streptozotocin-diabetic rats and age-matched controls. The stereological tool employed is the pair correlation function which is estimated by counting linear dipole probes of different sizes superimposed on ultrathin sections so as to be random in position and orientation. Unbiased estimates of the volume density of each glomerular component were estimated by point counting. Thereafter, estimates of the covariance and pair correlation function were determined from corresponding dipole counts. Plots of covariance and pair correlation functions against dipole length were almost identical in control and diabetic groups, indicating that diabetes did not disturb the normal spatial arrangements within glomeruli. However, differences were detected between compartments within groups. Whilst volume elements within all compartments were clustered at distances below about 8 μm (the approximate size of the basic cellular or other structural unit), the cluster size varied between compartments. The pattern was one of progressively smaller clusters in the sequence capillaries, podocytes, urinary space, mesangium. Beyond a distance of 8 μm, all glomerular components (in both control and diabetic groups) were arranged as expected for a 'random' (meaning neither clustered nor repulsed) volume process. These studies re-emphasize the relative invariance of biological organization and the value and limitations of covariance analysis for quantifying different levels of organization in different tissues and experimental groups.     

Coalignment of the muscle cell and nucleus,cell geometry and Vv in the tunica media of monkey cerebral arteries,by electron microscopy

We undertook to ascertain how well aligned is the rod-shaped nucleus within the spindle-shaped cell of vascular muscle in order that we might use the darkly staining nucleus in histological sections to indicate precisely the directional alignment of the cell. We fixed cerebral arteries from five monkeys under physiological pressure and embedded portions of the tissue so that mid-plane longitudinal sections of the arteries were obtained; the circumferentially arranged muscle cells were cut in cross-section. From the electron micrographs we obtained the cross-sectional profile of the cell and its nucleus, determining that the centre of the nucleus was on average 9·5 ± 5·8% (SD) away from the centre of the cell (expressed as a ratio of the cellular diameter). We calculated the alignment between the cell and nucleus to be from 0 to 3°, and obtained a volume fraction of 59% for muscle tissue in the tunica media of these arteries.     

Mesenchymal stem cells transplantation attenuates experimentally induced brain injury after neonatal hypoxia by different two routes of administrations

The neonatal hypoxic–ischemic encephalopathy (HIE) is an important cause of neurological morbidity and mortality in neonates. Cell therapy is considered a promising method for treating severe neurological disorders such as this one. Stem cells have the capacity for self-renewal and differentiation into certain cell lineages. The present study was aimed to find out the most beneficial route of bone marrow-derived mesenchymal stem cells (BMSCs) administration for the attenuation of experimentally induced HIE in neonatal rats. Sixty neonatal rats were divided randomly into four groups. Group 1: control group. Group 2: rats were exposed to bilateral ligation of cephalic arteries. Group 3: rats were exposed to bilateral ligation of cephalic arteries and then underwent intravenous (IV) BMSC injection. Group 4: rats were exposed to bilateral ligation of cephalic arteries and then underwent intracerebroventricular (ICV) BMSC injection. The animals were evaluated by (a) neurobehavioral tests; (b) histopathology, i.e., histological and immunohistochemical studies; and (3) gene expression studies. The BMSC treated groups (3 and 4) showed improvement in neurobehavioral tests, histopathological studies, and gene expression, as compared to non-injected lesioned rats (Group 2) with better improvement in Group 4 (ICV injections) than in Group 3 (IV injections).     

Effects of electromagnetic stirring and rare earth compounds on the microstructure and mechanical properties of hypereutectic Al–Si alloys

In this paper, the effects of rare earth addition and electromagnetic stirring on the microstructure and the mechanical properties of hypereutectic Al–Si alloys have been reported. Hypereutectic Al–Si alloy was prepared using liquid metallurgy route and modified with the addition of cerium oxide. To control the structure, slurry of hypereutectic Al–Si alloy was subjected to electromagnetic stirring before pouring into the mould. It was observed that the addition of cerium oxide (0.2 wt.%) refined the primary silicon particles and modified the eutectic silicon particles. Further, the electromagnetic stirring of the hypereutectic Al–Si alloy reduced the average size of primary silicon particles, from 152?±?9 to 120?±?6 μm, and the length of β-intermetallic compounds decreased from 314?±?12 to 234?±?10 μm. Similarly, the application of electromagnetic stirring on cerium oxide-modified hypereutectic Al–Si alloy also reduced the average size of primary silicon particles from 98?±?5 to 76?±?4 μm and the average length of β-intermetallic compounds from 225?±?7 to 203?±?5 μm. Mechanical properties namely tensile strength, ductility and hardness of the alloys were improved with electromagnetic stirring and addition of cerium oxide appreciably.     

Non‐Contact Evaluation of Osmosis‐Induced Shrinkage and Swelling Behavior of Articular Cartilage In‐Situ Using High‐Frequency Ultrasound

Abstract

The aim of the present study was to use high‐frequency ultrasound for the investigation of the transient osmosis‐induced free shrinkage‐swelling behaviors of normal articular cartilage in situ. Full‐thickness cartilage‐bone specimens were prepared from normal bovine patellae. The transient shrinkage and swelling strains of the cartilage induced by changing the bathing solution between physiological saline (0.15 M) and hypertonic saline (2 M) were monitored using a 50 MHz focused ultrasound beam. Both shrinkage and swelling strains showed temporary overshoots, followed by relaxation phases. The absolute peak value of the shrinkage strain (1.01%±0.62%) was significantly larger (p<0.05) than that of the swelling strain (0.40%±0.33%). It was found that the change of the mean ultrasound speed in cartilage could be approximately represented by an exponential function of time after the change of saline concentration.

This study successfully demonstrated that a high‐frequency focused ultrasound beam could be used to monitor the transient osmosis‐induced deformation of articular cartilage in a non‐contact way. Since the osmosis‐swelling behavior of cartilage relates to its compositional and structural characteristics and degeneration status, the reported ultrasound method may have potential for the characterization of cartilage degeneration, such as osteoarthritis.     

Morphometric,quantitative, and three‐dimensional analysis of the heart muscle fibers of old rats: Transmission electron microscopy and high‐resolution scanning electron microscopy methods

This research investigated the morphological, morphometric, and ultrastructural cardiomyocyte characteristics of male Wistar rats at 18 months of age. The animals were euthanized using an overdose of anesthesia (ketamine and xylazine, 150/10 mg/kg) and perfused transcardially, after which samples were collected for light microscopy, transmission electron microscopy, and high‐resolution scanning electron microscopy. The results showed that cardiomyocyte arrangement was disposed parallel between the mitochondria and the A‐, I‐, and H‐bands and their M‐ and Z‐lines from the sarcomere. The sarcomere junction areas had intercalated disks, a specific structure of heart muscle. The ultrastructural analysis revealed several mitochondria of various sizes and shapes intermingled between the blood capillaries and their endothelial cells; some red cells inside vessels are noted. The muscle cell sarcolemma could be observed associated with the described structures. The cardiomyocytes of old rats presented an average sarcomere length of 2.071 ± 0.09 μm, a mitochondrial volume density (Vv) of 0.3383, a mitochondrial average area of 0.537 ± 0.278 μm², a mitochondrial average length of 1.024 ± 0.352 μm, an average mitochondrial cristae thickness of 0.038 ± 0.09 μm and a ratio of mitochondrial greater length/lesser length of 1.929 ± 0.965. Of the observed mitochondrial shapes, 23.4% were rounded, 45.3% were elongated, and 31.1% had irregular profiles. In this study, we analyzed the morphology and morphometry of cardiomyocytes in old rats, focusing on mitochondria. These data are important for researchers who focus the changes in cardiac tissue, especially changes owing to pathologies and drug administration that may or may not be correlated with aging. Microsc. Res. Tech., 2013. © 2012 Wiley Periodicals, Inc.     

Morphological,microscopic, and physicochemical studies of Diospyros montana

Adulteration is the root cause of producing not only a chemically and pharmacologically inferior but also in some instances hazardous or poisonous drug. Despite availability of several techniques, microscopy and physicochemical analyses are the most practical approaches for crude drug authentication. Hence, the present study aimed to evaluate morphological, microscopic, and physicochemical properties of root, bark, leaf, and fruit of Diospyros montana Morphological properties were determined by sensory organs, whereas microscopic features of cross‐sections and powders were determined by light and scanning electron microscopy. The proximate and fluorescence analyses were performed using the standard guidelines. The physical examination of fresh, shade‐dried, and powdered material showed no significant change in color. The identifying cellular structures included cuboidal cork, pitted tracheids, scalariform, reticulate and spiral xylary vessels, and rosettes, raphide, and cuboidal calcium oxalate crystals. The stomatal number, stomatal index, vein‐islet and vein‐termination number, and palisade ratio in the leaf were found to be 293.91 ± 32.68 mm^?2, 64.18 ± 3.42%, 22.00 ± 3.81 mm^?2 and 38.40 ± 5.81 mm^?2, and 3.85 ± 0.60, respectively. Total ash, acid insoluble ash, water soluble ash and sulfated ash of leaf (9.00 ± 0.50%, 1.67 ± 0.23%, 2.00 ± 0.22% and 14.50 ± 0.99%, respectively), foaming index of bark and root (111.11 ± 2.11), and swelling index of fruit (19.00 ± 3.45) were higher than the other parts. The powder of different parts showed characteristic colors in the daylight and UV light upon treatment with various regents. The plant was found to be rich in saponins, fibers, and flavonoids. The results of the present study may serve as identifiers of different parts of Diospyros montana.