Objective: To collect high quality, representative tissue material from tumors and manage its distribution to different laboratories. Design: Prospective controlled study. Animals: Thirty-six dogs with mast cell tumor...Objective: To collect high quality, representative tissue material from tumors and manage its distribution to different laboratories. Design: Prospective controlled study. Animals: Thirty-six dogs with mast cell tumors. Procedures: The samples were submitted for the following analyses: stereology;histopathology;cell culture;breakdown for cytogenetic analysis of chromosomes (based on the Boxer breed published genome);Cell lysis for Real Time PCR and quantification of gene expression of CX 43, 32 and E-cadherin in canine mast cells. Results: Cytogenetic chromosome analysis, 90.9% of the samples were considered to be of good quality. For gene expression quantification of CX 43, 32 and E-cadherin in canine mast cell tumors (MCT), 95.5% of samples were considered to be of good quality. Conclusions and Clinical Relevance: We seek to assess the importance of surgical collection and post-surgical tissue preparation on laboratory testing by collecting surgical material appropriately to allow accurate diagnosis and reliable clinical prognosis and minimize errors caused by inadequate preparation of samples.展开更多
Aim: To evaluate the key lesions in spermatogenesis suppressed partially by testosterone undecanoate (TU) treatment. Methods: Adult male SD rats were treated with vehicle or TU (19 mg/kg) injection (i.m.) every 15 day...Aim: To evaluate the key lesions in spermatogenesis suppressed partially by testosterone undecanoate (TU) treatment. Methods: Adult male SD rats were treated with vehicle or TU (19 mg/kg) injection (i.m.) every 15 days for 130 days. The numbers of all types of cells (nuclei) in the seminiferous tubules and the interstitial tissue were estimated using a contemporary stereological tool, the optical disector. Results: In response to TU treatment, the numbers of non-type B spermatogonia, type B spermatogonia and late elongated spermatids per testis were reduced to 51 %, 66 % and 14 % of the controls, respectively. The conversion ratios from type B spermatogonia to early spermatocytes and pachytene spermatocytes were not significantly affected and the ratios to the later germ cell types fell to 51 % - 65 % of the controls. Less than 1.0 % of immature round spermatids were seen sloughing into the tubule lumen, 4.0 % of elongated spermatids retained in the seminiferous epithelium, and about half of the elongated spermatid nuclei appreciably malformed. Leydig cells were atrophied but their number and the peritubular myoid cell number per testis were unchanged. Conclusion: Double inhibition of spermatogenesis (i.e. inhibition at spermiation and spermatogonial conversion to type B spermatogonia), a scenario seen in the monkey and human following gona-dotrophin withdrawal, was not sufficiently effective for a complete spermatogenic suppression in the rat after TU treatment, probably due to ineffective inhibition of the Leydig cell population and therefore the intra-testicular testosterone levels.展开更多
Aim: To investigate the spermatozoal production rate of the testis and the spermatozoal storage capacity of the epi-didymis in monkeys and rats. Methods: The number of the late spermatids (steps 13 - 14 in the monkey ...Aim: To investigate the spermatozoal production rate of the testis and the spermatozoal storage capacity of the epi-didymis in monkeys and rats. Methods: The number of the late spermatids (steps 13 - 14 in the monkey or steps15 - 19 in the rat) per testis and the number of spermatozoa per epididymis were estimated in 6 normal adult monkeys(Macaca fascicularis) and 6 normal adult SD rats on 25 um-thick methacrylate-embedded sections using a contempo-rary unbiased and efficient stereological method-the optical disector. The diameter and length of the efferent ductulesand ductus epididymidis and the volume of the epididymal fluid in the tubules were also estimated. Results: The totalnumber of the late spermatids per testis was 2902 ±749 (million, x ±s) in the monkey, or 179 ±31 in the rat; thenumber of spermatozoa per epididymis was 3235 ±1835 in the monkey, or 241±76 in the rat. Conclusion: A largenumber of spermatozoa was densely packed and stored in the ductus epididymidis; the epididymal transit time for sper-matozoa was around 5 days in monkeys or 11 days in rats.展开更多
In the present study,tissue blocks were randomly sampled from the entire hippocampus of 6-week-old Long-Evans rats.Isotropic,uniform and random sections,60 nm thick,were prepared by isector.Fifteen fields of view were...In the present study,tissue blocks were randomly sampled from the entire hippocampus of 6-week-old Long-Evans rats.Isotropic,uniform and random sections,60 nm thick,were prepared by isector.Fifteen fields of view were randomly selected for each section and photographed using a transmission electron microscope.The mean internal and external diameters of the myelin sheaths were obtained by measuring the longest profile diameter perpendicular to its longest axis.The inner and outer perimeters of the myelin sheaths were estimated using the equidistant parallel test lines.The thickness of the myelin sheaths was estimated by direct orthogonal measurements in uniform,random locations.These stereological methods should permit an unbiased quantitative assessment of changes in the myelin sheaths of myelinated fibers in the hippocam-pus.展开更多
This study was organized to see whether vitamin E, as a strong antioxidant, could affect the abnormalities of testis structure caused by para-nonylphenol （p-NP） during its development. A total of 32 female Wistar ra...This study was organized to see whether vitamin E, as a strong antioxidant, could affect the abnormalities of testis structure caused by para-nonylphenol （p-NP） during its development. A total of 32 female Wistar rats after mating were divided into four groups （n = 8）： control, vitamin E （100 mg kg^-1 per day）, p-NP （250 mg kg^-1 per day） and p-NP ＋ vitamin E. The rats were treated from the seventh day of pregnancy till the twenty-first day. After weaning, the male pups were divided into the same groups and were treated orally for 90 days. Finally, the right testis was fixed, processed, stained and studied using stereological methods. The weight and volume of testis, volume of seminiferous tubules and its diameter, thickness of the basement membrane, height of the germinal epithelium, total number of types A and B spermatogonia, spermatocyte, spermatid and Sertoli cells were significantly reduced in p-NP group when compared with other groups. Co-administration of vitamin E and p-NP compensated for the adverse effects of p-NP on the above parameters. In addition, treatment with only vitamin E caused a significant increase in diameter, basement membrane thickness and height of germinal epithelium, number of spermatogonia and spermatocytes. Co-administration of vitamin E with p-NP could prevent the adverse effects ofp-NP on the testis structure during its development.展开更多
The evolution of both the metric and topological properties of the microstructure for Ni pow- der compacts at the late stages of sintering(V_v^s>0.90)has been investigated by means of stereological method.The effec...The evolution of both the metric and topological properties of the microstructure for Ni pow- der compacts at the late stages of sintering(V_v^s>0.90)has been investigated by means of stereological method.The effects of precompaction pressure and loose sintering have been stu- died and discussed.展开更多
AIM:To determine the histopathological changes of rifampicin applied intravitreally on retinal ganglion cells by means of stereological and histopathological methods.METHODS:For this study twenty-four New Zealand adul...AIM:To determine the histopathological changes of rifampicin applied intravitreally on retinal ganglion cells by means of stereological and histopathological methods.METHODS:For this study twenty-four New Zealand adult rabbits were divided into four groups(n=6 for each group).50μg/0.1mL(group 1),100μg/0.1mL(group 2),150μg/0.1mL(group 3) and 200μg/0.1mL(group 4),rifampicin were injected into the vitreous of the right eyes of animals,their left eyes were used as control(group 5).After the 28thday of application,animals were anesthetised with xylazine(8mg/kg,IM) and then their eyes were enucleated immediately.Patterns were taken away and eyes were prepared for both stereological and electromicroscopic observation.RESULTS:Depending on the high dose of rifampicin,some histopathological changes such as cytoplasmic dilatation and damaged membrane were observed on the electromicroscopic level.Using quantitative examination,which was done at the light microscopic level,it was shown that the number of neurons decreased linearly as rifampicin dose increased when compared with the control group.CONCLUSION:Based on these findings,low-dose rifampicin(50μg/0.1mL) may be useful for treatment of the ocular diseases.展开更多
The morphological and functional changes of the lungs in the early phase of blast injurywere analyzed and related quantitatively in rats with microcomputer-assisted morphometry.It wasfound that（1）significant or part...The morphological and functional changes of the lungs in the early phase of blast injurywere analyzed and related quantitatively in rats with microcomputer-assisted morphometry.It wasfound that（1）significant or partial correlation exists between stereological and functional indices,（2）changes of the endothelium and epithelium rather than the interstitium contribute to the increase ofharmonic and arithmetic thickness(T<sub>ht</sub>,T<sub>at</sub>)of the air-blood barrier,and（3）the regression equa-tions are as follows:（a）PaO<sub>2</sub>=3.56+1.62DL（O<sub>2</sub>）,r=0.678,P=0.032:（b）PaO<sub>2</sub>=2.99+3.49DL（O<sub>2</sub>）-0.0005S<sub>A</sub>+0.003S<sub>c</sub>-38.2V<sub>c</sub>-0.257T<sub>ap</sub>-0.028Dt（O<sub>2</sub>）,r<sub>y</sub>=0.9999,P=0.008;（c）PaCO<sub>2</sub>=7.69+0.0001S<sub>A</sub>-4.35T<sub>at</sub>+19.7T<sub>ht</sub>-46.2T<sub>hp</sub>-4.4T<sub>aen</sub>-5.87T<sub>aep</sub>,r<sub>y</sub>=0.9999,P=0.014;（d）pH=3.068+0.0005S<sub>A</sub>-1.79T<sub>at</sub>+12.2T<sub>ht</sub>-50.8T<sub>hp</sub>-2.52T<sub>aint</sub>+0.69DL（O<sub>2</sub>）,r<sub>y</sub>=0.9999,P=0.10.The significance and application of stereology in theestablishment of the quantitative relationship between morphological and functional indices are dis-cussed.Mathematical models to relate morphometric parameters to functional changes are possible.展开更多
This study was designed to investigate the relationship between hepatotoxicity of halothane and altered hepatic calcium homeostasis in enzyme-induced hypoxic rats by means of Ca2+ cytochemical location and EDX microan...This study was designed to investigate the relationship between hepatotoxicity of halothane and altered hepatic calcium homeostasis in enzyme-induced hypoxic rats by means of Ca2+ cytochemical location and EDX microanalysis combined with stereology of liver and ultrastructural analysis by computer. It was found that more calcium precipitated in the cytoplasm and mitochondria of liver than that in the controls following halothane exposure. This alteration appears to be the result of accelerated uptake of Ca2+ by hepatocytes and decreased output of loaded Ca2+ on the one hand, and increased release of reserved Ca2+ on the other hand. These results suggest that change in cullular Ca2+ balance plays an important role in the mechanism of hepatotoxicity induced by halothane exposure.展开更多
Objective To analyse quantitative changes of testicular peritubular cells in aging mice, and meanwhile investigate age-related changes in both Sertoli cells and germ cells with stereology. Methods Thirty male Kunming ...Objective To analyse quantitative changes of testicular peritubular cells in aging mice, and meanwhile investigate age-related changes in both Sertoli cells and germ cells with stereology. Methods Thirty male Kunming mice were enrolled in this study. The aged mice （n=15） were 18 months old. The young mice （n=15） were 6 months old, as the control. Haematoxylin and eosin staining was used to observe histological changes. Pathological evaluation rules of assignment scores were made to evaluate. Stereology techniques were applied to quantitatively analyse histological structure of the testicular tissues. Results Compared with young group, pathologic assignment scores of aging group were significantly higher （P〈0.05）. Diameter of seminiferous tubules in aging mice was atrophied significantly （P〈0.05）. Germ cell numerical density and Sertoli cell volume density of aging group were reduced significantly compared with young group （P〈0.05）. No significant differences were found in peritubular cell parameters compared with young group （P〉0.05）. Conclusion The pathological assignment scores suggest a succession of changes on the testis with age. The stereological numbers demonstrate damaged Sertoli cells and decreased germ cells, which is indicative of spermatogenic disturbance. However, no significant quantity changes of peritubular cells is observed in aging mice.展开更多
Sphere unfolding relationships are revisited with a specific focus on the analysis of segmented digital images of microstructures. Since the features of such images are most easily quantified by counting pixels, the r...Sphere unfolding relationships are revisited with a specific focus on the analysis of segmented digital images of microstructures. Since the features of such images are most easily quantified by counting pixels, the required equations are re-derived in terms of the histogram of areas (instead of diameters or radii) as inputs and it is shown that a substitution can be made that simplifies the calculation. A practical method is presented for utilizing negative number fraction bins (which sometimes arise from erroneous assumptions and/or insufficient numbers of observations) for the creation of error bars. The complete algorithm can be implemented in a spreadsheet. The derived unfolding equations are explored using both linear and logarithmic binning schemes, and the pros and cons of both binning schemes are illustrated using simulated data. The effects of the binning schemes on the stereological results are demonstrated and discussed with reference to their consequences for practical materials characterization situations, allowing for the suggestion of guidelines for proper application of this, and other, distribution-free stereological methods.展开更多
Background : Scleral fixation of intraocular lenses is a surgical technique that involves anchoring an artificial lens to the sclera. Traditional approaches, such as capsular bag placement, may not be feasible in cert...Background : Scleral fixation of intraocular lenses is a surgical technique that involves anchoring an artificial lens to the sclera. Traditional approaches, such as capsular bag placement, may not be feasible in certain situations, making scleral fixation a valuable alternative. The scleral reactions to different types of suture materials are not fully understood. Therefore, the present study describes the microscopic structure of normal scleral tissue and its changes with suture materials. Methods : We compared six groups of rabbit eyes focusing on the sclera: group with polytetrafluoroethylene( PTFE) chain, PTFE fiber, polypropylene( PPE) fiber and control groups. multilevel sampling and stereological methods were used for histological quantification of the leukocyte infiltration fractions and type Ⅰ and type Ⅲ collagen. Results : Quantitative histological evaluation revealed the following:(1) For all materials used, inflammation was present in the surrounding scleral tissue compared with healthy controls. However, leukocyte infiltration in the sclera was not statistically different between the materials.(2) As part of the evaluation of collagen, the greatest changes occurred in the PTFE fiber group at 2 weeks postoperatively. In the PTFE chain group, more significant changes were visible at 4 weeks.(3) The changes in the PPE fiber group compared to healthy scleral tissue were the least significant. Conclusions : From a histological point of view, it is evident that there are differences in the quantitative parameters between the untouched sclera and the sclera with suture material. Furthermore, distinctions were observed among various materials and across different time intervals.展开更多
Estimating fracture size is a fundamental aspect of rock engineering.However,determining the most probable diameter(MPD)from a fracture's surface trace remains challenging in the scientific community.The prevailin...Estimating fracture size is a fundamental aspect of rock engineering.However,determining the most probable diameter(MPD)from a fracture's surface trace remains challenging in the scientific community.The prevailing methodologies typically infer statistical distributions of fracture sizes rather than specific values.This research presents a novel approach to inferring the MPD and the true spatial distribution pattern of each fracture.The challenge lies in linking the inference process with the trace length of each fracture and the statistical characteristics of the entire outcrop.Additionally,it is necessary to address the non-unique inverse problem.The methodology comprises several key steps.Firstly,the issue of censoring bias is addressed by considering the lengths of the traces contained.Secondly,the orientation bias is corrected using the vector method,and the true mean trace length and standard deviation are estimated and derived.Thirdly,assuming a lognormal distribution for fracture sizes,the mean and standard deviation of diameters are derived through a high-order moment relationship between trace lengths and diameters,validated by Crofton's theorem.Finally,the MPDs of all trace samples are determined by relating MPDs to trace lengths and the standard deviation of diameters using stereology techniques.Furthermore,the true fracture spatial patterns are inverted based on spatial geometric relationships.The proposed methodology is validated through rigorous Monte Carlo simulation and applied in a practical engineering case study,demonstrating its potential for use in rock engineering applications.展开更多
Background:An affordable approach accepted worldwide to successfully treat mild maternal hyperglycemia in gestational diabetes(GDM),is diet-controlled therapy.As no elaborate research was available,this study was craf...Background:An affordable approach accepted worldwide to successfully treat mild maternal hyperglycemia in gestational diabetes(GDM),is diet-controlled therapy.As no elaborate research was available,this study was crafted to investigate morphometric stereological details and to determine the mean placental oxygen diffusion capacity for patients who were kept on diet therapy for mild gestational hyperglycemia.Methods:A clinical trial(NCT04907708)was conducted from January 2018 to February 2019.A total of 70 women completed the study,out of which 35 served as healthy controls as Group A and 35 were diagnosed as mild gestational diabetics according to WHO criteria(Group B).These patients were kept on a restricted calorie diet with light exercise during gestation and were followed regularly during gestation.Soon after delivery,conserved placentas underwent complete gross,microscopic,and stereological investigations with the point and intersection-counting methods.Results:Significant differences were observed for placental width and syncytial knots(p<0.010 and 0.025 respectively)between the groups on gross and light microscopy.Most of the parameters were non-significant,though numerically more in the GDM group.On stereological details,mean placental volume,mean placental components volumes(villi,inter-villous space,fetal capillary,and fetal connective tissue),mean villi and mean fetal capillary diameter,mean villi and capillary surface density and mean morphometric diffusing capacity of placenta showed non-significant results between the groups.Conclusion:Minimal changes were observed in gross,microscopy,and morphometric stereological details in the placentae of GDM patients managed with nutritional therapy during gestation compared with the healthy controls.展开更多
Objective:To determine the protective effect of co-enzyme Q10(CoQ10)on testicular tissue and sperm parameters in male rats treated with SunsetYellow FCF.Methods:Sixty male Sprague-Dawley rats were randomly divided int...Objective:To determine the protective effect of co-enzyme Q10(CoQ10)on testicular tissue and sperm parameters in male rats treated with SunsetYellow FCF.Methods:Sixty male Sprague-Dawley rats were randomly divided into 6 groups of the control,CoQ10(10 mg/kg/day),low dose of Sunset Yellow(2.5 mg/kg),high dose of Sunset Yellow(70 mg/kg),low dose of Sunset Yellow(2.5 mg/kg)plus CoQ10,and high dose of Sunset Yellow(70 mg/kg)plus CoQ10.The drugs were administered via daily oral gavages for 6 weeks.At the end of the experiment,sperm analysis,stereological and histological assessments of the testis were carried out.Results:The normal morphology(by 41.1%)and progressive spermatozoa(by 74.8%),testicle volume(by 33.4%),lumen volume(by 38.3%),interstitial tissue volume(by 44.7%),seminiferous tubule volume(by 40.7%),and number of spermatogonia(by 53.9%)and Leydig cells(by 70.7%)reduced in the rats that received high doses of Sunset Yellow in comparison to the control group.Nonetheless,all these alterations were recovered by CoQ10 treatment in the CoQ10 plus high dose of Sunset Yellow group.Furthermore,low doses of Sunset Yellow did not affect different parameters of the testis and sperm.Conclusions:CoQ10 could,to some extent,prevent structural changes of the testis induced by the high dose of SunsetYellow.展开更多
Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration. The goal of this study was to assess the effect on nerve regeneration, associating a hybrid chitosan me...Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration. The goal of this study was to assess the effect on nerve regeneration, associating a hybrid chitosan membrane with non-differentiated human mesenchymal stem cells isolated from Wharton＇s jelly of umbilical cord, in peripheral nerve reconstruction after crush injury. Chromosome analysis on human mesenchymal stem cell line from Wharton＇s jelly was carried out and no structural alterations were found in metaphase. Chitosan membranes were previously tested in vitro, to assess their ability in supporting human mesenchymal stem cell survival, expansion, and differentiation. For the in vivo testing, Sasco Sprague adult rats were divided in 4 groups of 6 or 7 animals each： Group 1, sciatic axonotmesis injury without any other intervention （Group 1-Crush）; Group 2, the axonotmesis lesion of 3 mm was infiltrated with a suspension of 1 250 -1 500 human mesenchymal stem cells （total volume of 50 pL） （Group 2-CrushCell）; Group 3, axonotmesis lesion of 3 mm was enwrapped with a chitosan type Ill membrane covered with a monolayer of non-differentiated human mesenchymal stem cells （Group 3-CrushChitlllCell） and Group 4, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane （Group 4-CrushChiUll）. Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index, static sciatic index, extensor postural thrust, and withdrawal reflex latency. Stereological analysis was carded out on regenerated nerve fibers. Results showed that infiltration of human mesenchymal stem cells, or the combination of chitosan membrane enwrapment and human mesenchymal stem cell enrichment after nerve crush injury provide a slight advantage to post-traumatic nerve regeneration. Results obtained with chitosan type III membrane alone confirmed that they significantly improve post-traumatic axonal regrowth and may represent a very promising clinical tool in peripheral nerve reconstructive surgery. Yet, umbilical cord human mesenchymal stem cells, that can be expanded in culture and induced to form several different types of cells, may prove, in future experiments, to be a new source of cells for cell therapy, including targets such as peripheral nerve and muscle.展开更多
Quantitative mineralogy has seen significant developments from the combination of scanning electron microscopy (SEM) with automatic image analysis and energy dispersive X-ray spectrometry (EDS). The mining industry is...Quantitative mineralogy has seen significant developments from the combination of scanning electron microscopy (SEM) with automatic image analysis and energy dispersive X-ray spectrometry (EDS). The mining industry is one of the fields that has benefited from this progress. In this paper, the authors present a newly developed quantitative method based on SEM-EDS and image analysis (IA), which is used to determine the mineralogical and environmental characteristics of mine tailings. The main objectives of the method are to be able to characterize sulphides and carbonates as monomineral particles, which control the acid generation from the tailings. Pure sulphides, calcite and quartz were blended to make mineralogical standards that represent typical mine tailings environmental behavior. The SEM-EDS-IA method achieved good mineralogical precision for medium (1-20 Wt%) and abundant (> 20 Wt%) minerals, with a relative error below 10 %. However, some corrections had to be applied to account for typical stereological effects (apparent particle diameter from polished surface) and preparation modes (particle segregation during resin hardening). Particle size analysis was used to calibrate the method and identify the corrections to be applied. Since mineralogical quantifications are based on the area of the observed particles, the most reliable particle size analyses (also obtained from particle area) typically lead to the best mineralogical characterization. However, the SEM based techniques may show some limitations for fine-grained particle quantification (< 10 μm), which required additional corrections. In this article, the technique is described, and it is applied to characterize fine-grained mine tailings with a size-by-size mineralogy (with sulphides and carbonates content). These results have been used by the Authors to propose an environmental management strategy for acid generating tailings using desulphurization by flotation.展开更多
Aim： To quantitatively study the histological changes of the testis and epididymis as a result of a drastic reduction of testosterone secretion. Methods： Fourteen adult Sprague-Dawley rats were injected intraperiton...Aim： To quantitatively study the histological changes of the testis and epididymis as a result of a drastic reduction of testosterone secretion. Methods： Fourteen adult Sprague-Dawley rats were injected intraperitoneally with ethane dimethane sulfonate （EDS, 75 mg/kg） and the same number of animals were injected with normal saline as a control. At days 7 and 12 （after treatment）, respectively, half of the animals from each group were killed. The testes and epididymides were removed and tissue blocks embedded in methacrylate resin. The cell number per testis was estimated using the stereological optical disector and some other parameters were obtained using other morphometric methods. Results： The EDS treatment resulted in an almost complete elimination of Leydig cells but had no effect on the numbers of Sertoli cells per testis. At day 7 after EDS treatment, many elongated spermatids were retained in the seminiferous epithelium and many round spermatids could be seen in the epididymal ducts. At day 12, a looser arrangement of spermatids and spermatocytes became evident, with apparent narrow empty spaces being formed between germ cells in an approximately radial direction towards the tubule lumen; the numbers （per testis） of non-type B spermatogonia and spermatocytes were similar to controls, whereas that of type B spermatogonia increased by 59%, and that of early round, elongating and late elongated spermatids decreased by 37%, 72% and 52%, respectively. Conclusion： The primary spermatogenic lesions following EDS administration were （i） spermiation failure and （ii） detachment of spermatids and spermatocytes associated with impairment in spermiogenesis and meiosis.展开更多
Aim: To study the effect of testosterone undecanoate (TU) injection on spermatogenesis in rats. Methods:Twenty adult SD rats received vehicle or TU (8 mg/kg, 19 mg/kg or 625 mg/kg) injection, im, every 15 days for 60d...Aim: To study the effect of testosterone undecanoate (TU) injection on spermatogenesis in rats. Methods:Twenty adult SD rats received vehicle or TU (8 mg/kg, 19 mg/kg or 625 mg/kg) injection, im, every 15 days for 60days, and another 38 animals received similar treatments for 130 days with half of them undergoing a recovery phase of120 days (5 rats for each treatment). At the end of the treatment, testes were removed and the diameter of the seminif-erous tubules and the number of late elongated spermatids (steps 15-19) per testis were estimated with stereologicalmethods as a measure of the spermatogenic efficiency. Results: Low dose (8 mg/kg) TU treatment virtually hadno effect on spermatogenesis. A dose of 19 mg/kg slightly suppressed spermatogenesis 60 days after treatment, and se-vere suppression occurred after another 70 days of dosing. Spermatogenesis was completely recovered at the end of therecovery phase. Large dose (625 mg/kg) TU treatment did not significantly affect spermatogenesis and was well toler-ated by animals. Conclusion: TU injection reversibly suppresses spermatogenesis in rats.展开更多
基金Supported by Fundacao de Amparo a Pesquisa do Estado de Sao Paulo(FAPESP).
文摘Objective: To collect high quality, representative tissue material from tumors and manage its distribution to different laboratories. Design: Prospective controlled study. Animals: Thirty-six dogs with mast cell tumors. Procedures: The samples were submitted for the following analyses: stereology;histopathology;cell culture;breakdown for cytogenetic analysis of chromosomes (based on the Boxer breed published genome);Cell lysis for Real Time PCR and quantification of gene expression of CX 43, 32 and E-cadherin in canine mast cells. Results: Cytogenetic chromosome analysis, 90.9% of the samples were considered to be of good quality. For gene expression quantification of CX 43, 32 and E-cadherin in canine mast cell tumors (MCT), 95.5% of samples were considered to be of good quality. Conclusions and Clinical Relevance: We seek to assess the importance of surgical collection and post-surgical tissue preparation on laboratory testing by collecting surgical material appropriately to allow accurate diagnosis and reliable clinical prognosis and minimize errors caused by inadequate preparation of samples.
文摘Aim: To evaluate the key lesions in spermatogenesis suppressed partially by testosterone undecanoate (TU) treatment. Methods: Adult male SD rats were treated with vehicle or TU (19 mg/kg) injection (i.m.) every 15 days for 130 days. The numbers of all types of cells (nuclei) in the seminiferous tubules and the interstitial tissue were estimated using a contemporary stereological tool, the optical disector. Results: In response to TU treatment, the numbers of non-type B spermatogonia, type B spermatogonia and late elongated spermatids per testis were reduced to 51 %, 66 % and 14 % of the controls, respectively. The conversion ratios from type B spermatogonia to early spermatocytes and pachytene spermatocytes were not significantly affected and the ratios to the later germ cell types fell to 51 % - 65 % of the controls. Less than 1.0 % of immature round spermatids were seen sloughing into the tubule lumen, 4.0 % of elongated spermatids retained in the seminiferous epithelium, and about half of the elongated spermatid nuclei appreciably malformed. Leydig cells were atrophied but their number and the peritubular myoid cell number per testis were unchanged. Conclusion: Double inhibition of spermatogenesis (i.e. inhibition at spermiation and spermatogonial conversion to type B spermatogonia), a scenario seen in the monkey and human following gona-dotrophin withdrawal, was not sufficiently effective for a complete spermatogenic suppression in the rat after TU treatment, probably due to ineffective inhibition of the Leydig cell population and therefore the intra-testicular testosterone levels.
文摘Aim: To investigate the spermatozoal production rate of the testis and the spermatozoal storage capacity of the epi-didymis in monkeys and rats. Methods: The number of the late spermatids (steps 13 - 14 in the monkey or steps15 - 19 in the rat) per testis and the number of spermatozoa per epididymis were estimated in 6 normal adult monkeys(Macaca fascicularis) and 6 normal adult SD rats on 25 um-thick methacrylate-embedded sections using a contempo-rary unbiased and efficient stereological method-the optical disector. The diameter and length of the efferent ductulesand ductus epididymidis and the volume of the epididymal fluid in the tubules were also estimated. Results: The totalnumber of the late spermatids per testis was 2902 ±749 (million, x ±s) in the monkey, or 179 ±31 in the rat; thenumber of spermatozoa per epididymis was 3235 ±1835 in the monkey, or 241±76 in the rat. Conclusion: A largenumber of spermatozoa was densely packed and stored in the ductus epididymidis; the epididymal transit time for sper-matozoa was around 5 days in monkeys or 11 days in rats.
基金the National Natural Science Foundation of China,No.30973155
文摘In the present study,tissue blocks were randomly sampled from the entire hippocampus of 6-week-old Long-Evans rats.Isotropic,uniform and random sections,60 nm thick,were prepared by isector.Fifteen fields of view were randomly selected for each section and photographed using a transmission electron microscope.The mean internal and external diameters of the myelin sheaths were obtained by measuring the longest profile diameter perpendicular to its longest axis.The inner and outer perimeters of the myelin sheaths were estimated using the equidistant parallel test lines.The thickness of the myelin sheaths was estimated by direct orthogonal measurements in uniform,random locations.These stereological methods should permit an unbiased quantitative assessment of changes in the myelin sheaths of myelinated fibers in the hippocam-pus.
文摘This study was organized to see whether vitamin E, as a strong antioxidant, could affect the abnormalities of testis structure caused by para-nonylphenol （p-NP） during its development. A total of 32 female Wistar rats after mating were divided into four groups （n = 8）： control, vitamin E （100 mg kg^-1 per day）, p-NP （250 mg kg^-1 per day） and p-NP ＋ vitamin E. The rats were treated from the seventh day of pregnancy till the twenty-first day. After weaning, the male pups were divided into the same groups and were treated orally for 90 days. Finally, the right testis was fixed, processed, stained and studied using stereological methods. The weight and volume of testis, volume of seminiferous tubules and its diameter, thickness of the basement membrane, height of the germinal epithelium, total number of types A and B spermatogonia, spermatocyte, spermatid and Sertoli cells were significantly reduced in p-NP group when compared with other groups. Co-administration of vitamin E and p-NP compensated for the adverse effects of p-NP on the above parameters. In addition, treatment with only vitamin E caused a significant increase in diameter, basement membrane thickness and height of germinal epithelium, number of spermatogonia and spermatocytes. Co-administration of vitamin E with p-NP could prevent the adverse effects ofp-NP on the testis structure during its development.
文摘The evolution of both the metric and topological properties of the microstructure for Ni pow- der compacts at the late stages of sintering(V_v^s>0.90)has been investigated by means of stereological method.The effects of precompaction pressure and loose sintering have been stu- died and discussed.
文摘AIM:To determine the histopathological changes of rifampicin applied intravitreally on retinal ganglion cells by means of stereological and histopathological methods.METHODS:For this study twenty-four New Zealand adult rabbits were divided into four groups(n=6 for each group).50μg/0.1mL(group 1),100μg/0.1mL(group 2),150μg/0.1mL(group 3) and 200μg/0.1mL(group 4),rifampicin were injected into the vitreous of the right eyes of animals,their left eyes were used as control(group 5).After the 28thday of application,animals were anesthetised with xylazine(8mg/kg,IM) and then their eyes were enucleated immediately.Patterns were taken away and eyes were prepared for both stereological and electromicroscopic observation.RESULTS:Depending on the high dose of rifampicin,some histopathological changes such as cytoplasmic dilatation and damaged membrane were observed on the electromicroscopic level.Using quantitative examination,which was done at the light microscopic level,it was shown that the number of neurons decreased linearly as rifampicin dose increased when compared with the control group.CONCLUSION:Based on these findings,low-dose rifampicin(50μg/0.1mL) may be useful for treatment of the ocular diseases.
基金This study was supported by the National Natural Sciences Foundation of China
文摘The morphological and functional changes of the lungs in the early phase of blast injurywere analyzed and related quantitatively in rats with microcomputer-assisted morphometry.It wasfound that（1）significant or partial correlation exists between stereological and functional indices,（2）changes of the endothelium and epithelium rather than the interstitium contribute to the increase ofharmonic and arithmetic thickness(T<sub>ht</sub>,T<sub>at</sub>)of the air-blood barrier,and（3）the regression equa-tions are as follows:（a）PaO<sub>2</sub>=3.56+1.62DL（O<sub>2</sub>）,r=0.678,P=0.032:（b）PaO<sub>2</sub>=2.99+3.49DL（O<sub>2</sub>）-0.0005S<sub>A</sub>+0.003S<sub>c</sub>-38.2V<sub>c</sub>-0.257T<sub>ap</sub>-0.028Dt（O<sub>2</sub>）,r<sub>y</sub>=0.9999,P=0.008;（c）PaCO<sub>2</sub>=7.69+0.0001S<sub>A</sub>-4.35T<sub>at</sub>+19.7T<sub>ht</sub>-46.2T<sub>hp</sub>-4.4T<sub>aen</sub>-5.87T<sub>aep</sub>,r<sub>y</sub>=0.9999,P=0.014;（d）pH=3.068+0.0005S<sub>A</sub>-1.79T<sub>at</sub>+12.2T<sub>ht</sub>-50.8T<sub>hp</sub>-2.52T<sub>aint</sub>+0.69DL（O<sub>2</sub>）,r<sub>y</sub>=0.9999,P=0.10.The significance and application of stereology in theestablishment of the quantitative relationship between morphological and functional indices are dis-cussed.Mathematical models to relate morphometric parameters to functional changes are possible.
文摘This study was designed to investigate the relationship between hepatotoxicity of halothane and altered hepatic calcium homeostasis in enzyme-induced hypoxic rats by means of Ca2+ cytochemical location and EDX microanalysis combined with stereology of liver and ultrastructural analysis by computer. It was found that more calcium precipitated in the cytoplasm and mitochondria of liver than that in the controls following halothane exposure. This alteration appears to be the result of accelerated uptake of Ca2+ by hepatocytes and decreased output of loaded Ca2+ on the one hand, and increased release of reserved Ca2+ on the other hand. These results suggest that change in cullular Ca2+ balance plays an important role in the mechanism of hepatotoxicity induced by halothane exposure.
文摘Objective To analyse quantitative changes of testicular peritubular cells in aging mice, and meanwhile investigate age-related changes in both Sertoli cells and germ cells with stereology. Methods Thirty male Kunming mice were enrolled in this study. The aged mice （n=15） were 18 months old. The young mice （n=15） were 6 months old, as the control. Haematoxylin and eosin staining was used to observe histological changes. Pathological evaluation rules of assignment scores were made to evaluate. Stereology techniques were applied to quantitatively analyse histological structure of the testicular tissues. Results Compared with young group, pathologic assignment scores of aging group were significantly higher （P〈0.05）. Diameter of seminiferous tubules in aging mice was atrophied significantly （P〈0.05）. Germ cell numerical density and Sertoli cell volume density of aging group were reduced significantly compared with young group （P〈0.05）. No significant differences were found in peritubular cell parameters compared with young group （P〉0.05）. Conclusion The pathological assignment scores suggest a succession of changes on the testis with age. The stereological numbers demonstrate damaged Sertoli cells and decreased germ cells, which is indicative of spermatogenic disturbance. However, no significant quantity changes of peritubular cells is observed in aging mice.
文摘Sphere unfolding relationships are revisited with a specific focus on the analysis of segmented digital images of microstructures. Since the features of such images are most easily quantified by counting pixels, the required equations are re-derived in terms of the histogram of areas (instead of diameters or radii) as inputs and it is shown that a substitution can be made that simplifies the calculation. A practical method is presented for utilizing negative number fraction bins (which sometimes arise from erroneous assumptions and/or insufficient numbers of observations) for the creation of error bars. The complete algorithm can be implemented in a spreadsheet. The derived unfolding equations are explored using both linear and logarithmic binning schemes, and the pros and cons of both binning schemes are illustrated using simulated data. The effects of the binning schemes on the stereological results are demonstrated and discussed with reference to their consequences for practical materials characterization situations, allowing for the suggestion of guidelines for proper application of this, and other, distribution-free stereological methods.
基金supported by the Charles University Cooperatio Program,Research Areas MED/DIAG,and Surgical Disciplinessupport from the Ministry of Education,Youth,and Sports of the Czech Republic under Project FIND No.CZ.02.1.01/0.0/0.0/16_019/0000787.L.V.was also supported by Charles University SVV-2025260773.
文摘Background : Scleral fixation of intraocular lenses is a surgical technique that involves anchoring an artificial lens to the sclera. Traditional approaches, such as capsular bag placement, may not be feasible in certain situations, making scleral fixation a valuable alternative. The scleral reactions to different types of suture materials are not fully understood. Therefore, the present study describes the microscopic structure of normal scleral tissue and its changes with suture materials. Methods : We compared six groups of rabbit eyes focusing on the sclera: group with polytetrafluoroethylene( PTFE) chain, PTFE fiber, polypropylene( PPE) fiber and control groups. multilevel sampling and stereological methods were used for histological quantification of the leukocyte infiltration fractions and type Ⅰ and type Ⅲ collagen. Results : Quantitative histological evaluation revealed the following:(1) For all materials used, inflammation was present in the surrounding scleral tissue compared with healthy controls. However, leukocyte infiltration in the sclera was not statistically different between the materials.(2) As part of the evaluation of collagen, the greatest changes occurred in the PTFE fiber group at 2 weeks postoperatively. In the PTFE chain group, more significant changes were visible at 4 weeks.(3) The changes in the PPE fiber group compared to healthy scleral tissue were the least significant. Conclusions : From a histological point of view, it is evident that there are differences in the quantitative parameters between the untouched sclera and the sclera with suture material. Furthermore, distinctions were observed among various materials and across different time intervals.
基金supported by the National Natural Science Foundation of China(Grant Nos.41941017 and U1702241).
文摘Estimating fracture size is a fundamental aspect of rock engineering.However,determining the most probable diameter(MPD)from a fracture's surface trace remains challenging in the scientific community.The prevailing methodologies typically infer statistical distributions of fracture sizes rather than specific values.This research presents a novel approach to inferring the MPD and the true spatial distribution pattern of each fracture.The challenge lies in linking the inference process with the trace length of each fracture and the statistical characteristics of the entire outcrop.Additionally,it is necessary to address the non-unique inverse problem.The methodology comprises several key steps.Firstly,the issue of censoring bias is addressed by considering the lengths of the traces contained.Secondly,the orientation bias is corrected using the vector method,and the true mean trace length and standard deviation are estimated and derived.Thirdly,assuming a lognormal distribution for fracture sizes,the mean and standard deviation of diameters are derived through a high-order moment relationship between trace lengths and diameters,validated by Crofton's theorem.Finally,the MPDs of all trace samples are determined by relating MPDs to trace lengths and the standard deviation of diameters using stereology techniques.Furthermore,the true fracture spatial patterns are inverted based on spatial geometric relationships.The proposed methodology is validated through rigorous Monte Carlo simulation and applied in a practical engineering case study,demonstrating its potential for use in rock engineering applications.
文摘Background:An affordable approach accepted worldwide to successfully treat mild maternal hyperglycemia in gestational diabetes(GDM),is diet-controlled therapy.As no elaborate research was available,this study was crafted to investigate morphometric stereological details and to determine the mean placental oxygen diffusion capacity for patients who were kept on diet therapy for mild gestational hyperglycemia.Methods:A clinical trial(NCT04907708)was conducted from January 2018 to February 2019.A total of 70 women completed the study,out of which 35 served as healthy controls as Group A and 35 were diagnosed as mild gestational diabetics according to WHO criteria(Group B).These patients were kept on a restricted calorie diet with light exercise during gestation and were followed regularly during gestation.Soon after delivery,conserved placentas underwent complete gross,microscopic,and stereological investigations with the point and intersection-counting methods.Results:Significant differences were observed for placental width and syncytial knots(p<0.010 and 0.025 respectively)between the groups on gross and light microscopy.Most of the parameters were non-significant,though numerically more in the GDM group.On stereological details,mean placental volume,mean placental components volumes(villi,inter-villous space,fetal capillary,and fetal connective tissue),mean villi and mean fetal capillary diameter,mean villi and capillary surface density and mean morphometric diffusing capacity of placenta showed non-significant results between the groups.Conclusion:Minimal changes were observed in gross,microscopy,and morphometric stereological details in the placentae of GDM patients managed with nutritional therapy during gestation compared with the healthy controls.
文摘Objective:To determine the protective effect of co-enzyme Q10(CoQ10)on testicular tissue and sperm parameters in male rats treated with SunsetYellow FCF.Methods:Sixty male Sprague-Dawley rats were randomly divided into 6 groups of the control,CoQ10(10 mg/kg/day),low dose of Sunset Yellow(2.5 mg/kg),high dose of Sunset Yellow(70 mg/kg),low dose of Sunset Yellow(2.5 mg/kg)plus CoQ10,and high dose of Sunset Yellow(70 mg/kg)plus CoQ10.The drugs were administered via daily oral gavages for 6 weeks.At the end of the experiment,sperm analysis,stereological and histological assessments of the testis were carried out.Results:The normal morphology(by 41.1%)and progressive spermatozoa(by 74.8%),testicle volume(by 33.4%),lumen volume(by 38.3%),interstitial tissue volume(by 44.7%),seminiferous tubule volume(by 40.7%),and number of spermatogonia(by 53.9%)and Leydig cells(by 70.7%)reduced in the rats that received high doses of Sunset Yellow in comparison to the control group.Nonetheless,all these alterations were recovered by CoQ10 treatment in the CoQ10 plus high dose of Sunset Yellow group.Furthermore,low doses of Sunset Yellow did not affect different parameters of the testis and sperm.Conclusions:CoQ10 could,to some extent,prevent structural changes of the testis induced by the high dose of SunsetYellow.
基金supported by Technology and Science Foundation(FCT),Education and Science Ministry,Portugal,through the financed research project PTDC/DES/104036/2008by QREN N°1372-Nucleus I&DT for the Development of Products for Regenerative Medicine and Cell Therapies-Núcleo Biomat&CellAndrea Grtner has a Doctoral Grantfrom Technology and Science Foundation(FCT),Education and Science Ministry,Portugal,SFRH/BD/70211/2010
文摘Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration. The goal of this study was to assess the effect on nerve regeneration, associating a hybrid chitosan membrane with non-differentiated human mesenchymal stem cells isolated from Wharton＇s jelly of umbilical cord, in peripheral nerve reconstruction after crush injury. Chromosome analysis on human mesenchymal stem cell line from Wharton＇s jelly was carried out and no structural alterations were found in metaphase. Chitosan membranes were previously tested in vitro, to assess their ability in supporting human mesenchymal stem cell survival, expansion, and differentiation. For the in vivo testing, Sasco Sprague adult rats were divided in 4 groups of 6 or 7 animals each： Group 1, sciatic axonotmesis injury without any other intervention （Group 1-Crush）; Group 2, the axonotmesis lesion of 3 mm was infiltrated with a suspension of 1 250 -1 500 human mesenchymal stem cells （total volume of 50 pL） （Group 2-CrushCell）; Group 3, axonotmesis lesion of 3 mm was enwrapped with a chitosan type Ill membrane covered with a monolayer of non-differentiated human mesenchymal stem cells （Group 3-CrushChitlllCell） and Group 4, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane （Group 4-CrushChiUll）. Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index, static sciatic index, extensor postural thrust, and withdrawal reflex latency. Stereological analysis was carded out on regenerated nerve fibers. Results showed that infiltration of human mesenchymal stem cells, or the combination of chitosan membrane enwrapment and human mesenchymal stem cell enrichment after nerve crush injury provide a slight advantage to post-traumatic nerve regeneration. Results obtained with chitosan type III membrane alone confirmed that they significantly improve post-traumatic axonal regrowth and may represent a very promising clinical tool in peripheral nerve reconstructive surgery. Yet, umbilical cord human mesenchymal stem cells, that can be expanded in culture and induced to form several different types of cells, may prove, in future experiments, to be a new source of cells for cell therapy, including targets such as peripheral nerve and muscle.
文摘Quantitative mineralogy has seen significant developments from the combination of scanning electron microscopy (SEM) with automatic image analysis and energy dispersive X-ray spectrometry (EDS). The mining industry is one of the fields that has benefited from this progress. In this paper, the authors present a newly developed quantitative method based on SEM-EDS and image analysis (IA), which is used to determine the mineralogical and environmental characteristics of mine tailings. The main objectives of the method are to be able to characterize sulphides and carbonates as monomineral particles, which control the acid generation from the tailings. Pure sulphides, calcite and quartz were blended to make mineralogical standards that represent typical mine tailings environmental behavior. The SEM-EDS-IA method achieved good mineralogical precision for medium (1-20 Wt%) and abundant (> 20 Wt%) minerals, with a relative error below 10 %. However, some corrections had to be applied to account for typical stereological effects (apparent particle diameter from polished surface) and preparation modes (particle segregation during resin hardening). Particle size analysis was used to calibrate the method and identify the corrections to be applied. Since mineralogical quantifications are based on the area of the observed particles, the most reliable particle size analyses (also obtained from particle area) typically lead to the best mineralogical characterization. However, the SEM based techniques may show some limitations for fine-grained particle quantification (< 10 μm), which required additional corrections. In this article, the technique is described, and it is applied to characterize fine-grained mine tailings with a size-by-size mineralogy (with sulphides and carbonates content). These results have been used by the Authors to propose an environmental management strategy for acid generating tailings using desulphurization by flotation.
文摘Aim： To quantitatively study the histological changes of the testis and epididymis as a result of a drastic reduction of testosterone secretion. Methods： Fourteen adult Sprague-Dawley rats were injected intraperitoneally with ethane dimethane sulfonate （EDS, 75 mg/kg） and the same number of animals were injected with normal saline as a control. At days 7 and 12 （after treatment）, respectively, half of the animals from each group were killed. The testes and epididymides were removed and tissue blocks embedded in methacrylate resin. The cell number per testis was estimated using the stereological optical disector and some other parameters were obtained using other morphometric methods. Results： The EDS treatment resulted in an almost complete elimination of Leydig cells but had no effect on the numbers of Sertoli cells per testis. At day 7 after EDS treatment, many elongated spermatids were retained in the seminiferous epithelium and many round spermatids could be seen in the epididymal ducts. At day 12, a looser arrangement of spermatids and spermatocytes became evident, with apparent narrow empty spaces being formed between germ cells in an approximately radial direction towards the tubule lumen; the numbers （per testis） of non-type B spermatogonia and spermatocytes were similar to controls, whereas that of type B spermatogonia increased by 59%, and that of early round, elongating and late elongated spermatids decreased by 37%, 72% and 52%, respectively. Conclusion： The primary spermatogenic lesions following EDS administration were （i） spermiation failure and （ii） detachment of spermatids and spermatocytes associated with impairment in spermiogenesis and meiosis.
基金Financially supported by a"9th five-year" National Key Grant of Science and Technology (Grant number:969040401),and by Sichuan Committee of Education.
文摘Aim: To study the effect of testosterone undecanoate (TU) injection on spermatogenesis in rats. Methods:Twenty adult SD rats received vehicle or TU (8 mg/kg, 19 mg/kg or 625 mg/kg) injection, im, every 15 days for 60days, and another 38 animals received similar treatments for 130 days with half of them undergoing a recovery phase of120 days (5 rats for each treatment). At the end of the treatment, testes were removed and the diameter of the seminif-erous tubules and the number of late elongated spermatids (steps 15-19) per testis were estimated with stereologicalmethods as a measure of the spermatogenic efficiency. Results: Low dose (8 mg/kg) TU treatment virtually hadno effect on spermatogenesis. A dose of 19 mg/kg slightly suppressed spermatogenesis 60 days after treatment, and se-vere suppression occurred after another 70 days of dosing. Spermatogenesis was completely recovered at the end of therecovery phase. Large dose (625 mg/kg) TU treatment did not significantly affect spermatogenesis and was well toler-ated by animals. Conclusion: TU injection reversibly suppresses spermatogenesis in rats.
