Tamoxifen induced multinucleated cells (symplasts) and distortion of seminiferous tubules in rat testis

Aim: To evaluate the effect of tamoxifen citrate on male reproductive system of rat. Methods: Groups of male rats were gavaged with tamoxifen at doses of 200 mg·kg-1·d-1, 400 mg·kg-1·d-1 or 800 mg·kg-1·d-1 in 0.1 mL olive oil for 10 consecutive days. Controls were treated with 0.1 mL olive oil. Rats were anesthetized and killed on d 3, d 15 or d 35 after the last dose. Testes were collected, processed for paraffin embedding, sectioned at 5 μm thickness, stained with H&E and analyzed microscopically. Results: There was a dose-dependent increase in the occurrence of seminiferous tubular distortion with germinal cell sloughing. The highest dose increased the number of multinucleated giant cells on d 3 and d 15. Conclusion: Tamoxifen citrate induces multinucleated giant cells and germinal epithelial sloughing in a dose-dependent manner and these changes are detrimental to male fertility.

EFFECT OF DENTIN TUBULES ON THE MECHANICAL PROPERTIES OF DENTIN. PART Ⅰ: STRESS-STRAIN RELATIONS AND STRENGTH CRITERION

As known, there is a large number of dentin tubules in dentin. These tubules have varying radii and are shaped into radially parallel pattern. The anisotropy of microstructure of dentin shows that dentin should be treated as a ma- terial of varying transverse isotropy. In this Part, the elastic stress-strain relations and the quadratic strength criterion are established in the form of having varying transverse isotropy, in the framework of micromechanics to take into account of the effect of the microstructures-dentin tubules. Simplified forms for isotropic and ho- mogeneous cases, as well as the corresponding plane stress form of the stress-strain relations are also given. These theoretical models are very well supported by the experiments shown later in the continued paper (Part Ⅱ).

EFFECT OF DENTIN TUBULES ON THE MECHANICAL PROPERTIES OF DENTIN PART Ⅲ: NUMERICAL ANALYSIS

Imitating a real tooth and the periodontal supporting tissues, we have established a 2D finite element model and carried out a numerical analysis based on the inhomogeneous and anisotropic (IA) stress-strain relation and strength model of dentin proposed in the preceding Parts Ⅰ and Ⅱ, and the conventional homogeneous and isotropic (HI) model, respectively. Quite a few cases of loadings for a non-defected and a defected tooth are considered. The numerical results show that the stress level predicted by the IA model is remarkably higher than that by the HI model, revealing that the effect of the dentin tubules should be taken into a serious consideration from the viewpoint of biomechanics.

EFFECT OF DENTIN TUBULES TO THE MECHANICAL PROPERTIES OF DENTIN.PART Ⅱ:EXPERIMENTAL STUDY

To verify the theoretical models of varying transversely isotropic stress-strain relations of dentin established in the preceding work(Part Ⅰ),we per- form a set of experiments.Because of the very fine tooth size,it usually seems to be difficult to directly measure the inhomogeneous and anisotropic parameters of dentin.In this paper,by the digital speckle correlation method,tensile experiments are made on the small dentin samples either parallel or perpendicular to the dentin tubules.With the theoretically predicted elastic stress-strain relations,an optimiza- tion method is proposed to fit the strain curve adapted to the experimental data. The results show that the theoretical elastic stress-strain relations coincides very well with the experimental observations.The determined Young's modulus and Poisson's ratio of dentin matrix are 29.5GPa and 0.44,respectively,in the optimization sense.

Actin phosphorylation correlates with actin sequestration in ATP-depleted rabbit renal proximal tubules

Objective: To demonstrate the relation sh ip between actin phosphorylation and actin sequestration in ATP-depleted rabbi t renal proximal tubules. Methods: Using two-dimensional electr ophoreses and Western blotting to analyze the phosphorylation state of the seque stered actin in rabbit renal proximal tubules. Results: The anal ytical result of the sequestered actin indicated that nearly half of the actin w as phosphorylated on serine residue(s). Conclusion: Result sugge sted a close correlation between actin sequestration and actin phosphorylation i n ATP-depleted rabbit renal proximal tubules.

Effect of Different Irradiation Times on the Occlusion of Dentinal Tubules When Using a Nd:YAG Laser: An <i>in Vitro</i>SEM Study

Objective: To evaluate the effect of different irradiation times on the occlusion of dentinal tubules when using Nd:YAG laser. Background data: Dentin hypersensitivity is a frequent problem that has limited treatment success despite many chemical and physical therapies. Methods: Four coronal dentin disks 2 mm thick were cut with a low-speed diamond saw from four freshly extracted intact first molars. The coronal dentin surface of each disk was divided into four regions, each approximately 2 mm × 3 mm. The dentin surfaces were treated with 27% EDTA then the four regions irradiated separately in a randomized pattern with a Nd:YAG laser (120 mJ, 5 Hz), using irradiation times of 0 s, 20 s, 40 s and 60 s, representing laser energies of 0 J/cm2, 200 J/cm2, 400 J/cm2 and 600 J/cm2, respectively. SEM photomicro-graphs were taken at 1500× and 3000× to calculate the dentinal tubule orifice occlusion rates and to measure the tubule diameters, respectively. Results: For the laser irradiation times of 0 s, 20 s, 40 s and 60 s, the corresponding dentinal tubule occlusion rates were 2.05 (SD 0.29)%, 10.01 (1.71)%, 23.58 (2.51)% and 90.23 (2.24)%, respectively;and the tubule diameters were 4.18 (0.32) μm, 3.46 (0.24) μm, 1.69 (0.32) μm and 0.12 (0.02) μm, respectively. There were significant differences among all groups for both measured variables (p ≤ 0.005). Conclusions: Within the limitations of this in Vitro study, when using a Nd:YAG laser at 120 mJ and 5 Hz, an irradiation time of 60 s achieved the best sealing of the coronal dentinal tubule orifices.

MORPHOLOGICAL TRANSITION BETWEEN VESICLES AND TUBULES FOR A GLYCOPOLYMER-CONTAINING AMPHIPHILIC DIBLOCK COPOLYMER IN AQUEOUS SOLUTIONS

The morphological transition of molecular assemblies in aqueous solutions for a new amphiphilic diblock copolymer induced by changing the initial solvent conditions was studied by transmission electron microscopy (TEM). The copolymer was polystyrene(77)-b-poly[2-(beta -D-glucopyranosyloxy)ethyl acrylate (6)] (PSt(77)-b-PGEA(6)) and the solvent was a mixture of DMF and THF. PSt(77)-b-PGEA(6) yields vesicles and tubules when it is initially dissolved in THF and DMF respectively. The morphological transition between vesicles and tubules can be achieved by simply varying the amounts of THF and DMF, or changing the temperature at which the aggregates were prepared.

The mechanism of dentine hypersensitivity:Stimuli-induced directional cation transport through dentinal tubules

Dentine hypersensitivity is an annoying worldwide disease,yet its mechanism remains unclear.The long-used hydrodynamic theory,a stimuli-induced fluid-flow process,describes the pain processes.However,no experimental evidence supports the statements.Here,we demonstrate that stimuli-induced directional cation transport,rather than fluid-flow,through dentinal tubules actually leads to dentine hypersensitivity.The in vitro/in vivo electro-chemical and electro-neurophysiological approaches reveal the cation current through the nanoconfined negatively charged dentinal tubules coming from external stimuli(pressure,pH,and temperature)on dentin surface and further triggering the nerve impulses causing the dentine hypersensitivity.Furthermore,the cationic-hydrogels blocked dentinal tubules could significantly reduce the stimuli-triggered nerve action potentials and the anionhydrogels counterpart enhances those,supporting the cation-flow transducing dentine hypersensitivity.Therefore,the inspired ion-blocking desensitizing therapies have achieved remarkable pain relief in clinical applications.The proposed mechanism would enrich the basic knowledge of dentistry and further foster breakthrough initiatives in hypersensitivity mitigation and cure.

Differential Notch Activity Is Required for Homeostasis of Malpighian Tubules in Adult Drosophila

Precise regulation of stem cell self-renewal versus differentiation is important for the maintenance of tissue ho- meostasis. Early loss of stem cell activity results in premature ageing, while excess stem cell activity leads to over- proliferation and cancer initiation （Fuchs et al., 2004; Scadden, 2006; Jones and Wagers, 2008; Morrison and Spradling, 2008）. Thus, understanding the mechanism con- trolling stem cell self-renewal and differentiation is critical for development of potential clinic therapy in the future.

Comparative transcriptomic analysis of larval and adult Malpighian tubules from the cotton bollworm Helicoverpa armigera

Malpighian tubules (MTs)are moregulatory organs of insects.However, usually considered the key excretory and os- increasing evidence has suggested that MTs perform many more functions than just osmoregulation.Until now,the molecular and physiological functions of MTs in the cotton bollworm (Helicoverpa armigera),a very important agricultural pest,are largely unknown.In this study,the transcriptomes of H. armigera MTs from larvae,male adults and female adults were sequenced using RNA-Seq technology,and comparative analyses of transcriptomes between two life stages (larval and adult)and between adult sexes were conducted.We generated a total of 84 643 high-quality unigenes,and identified a large number of abundant transcripts putatively encoding proteins involved in diuresis,detoxification,immunity,carbohydrate transport and metabolism,development and reproduction.We found that the expression pattern of unigenes was relatively similar between female and male adult MTs,but different between larval and adult MTs.Our data suggest that insect MTs may take multiple physiological fimctions as versatile organs.The extensive alterations in gene expression in MTs occurred from larvae to adults reflect an ecological adaptation to different feeding habits.Sexual dimorphism in the cotton bollworm is somewhat indicated by the transcriptional difference of genes related to carbohydrate metabolism,detoxification,immunity and reproduction in the MTs of male and female adults.

STUDY ON MICROTUBULES IN SPERMIOGENESIS OF THE DOMESTIC DUCKS

Ⅰ. INTRODUCTION Ever since Sabatini improved on the fixing method for the electron microscope with glutaradehyde instead of osmic acid and permanganate potassinm, the microtubules have been generally found in eukaryocytes, as reported by Tilney and Heplar et al. A microtubule is a fine tubule of about 210—270 in diameter and it is an important cell organelle, supporting the cells and making them move. It is said to be 'the

Study on Transplanting Germ Like Cells Differentiated from Embryonic Stem Cell into Mouse Seminiferous Tubules

Objective To investigate whether germ like cells isolated from embryoid body formed by mouse embryonic stem cells could survive and initiate spermatogenesis in seminiferous tubules of adult mice. Methods SSEA-1＋ cells were isolated from embryoid bodies prepared from mouse EGFP-ES cells, after retinoic acid treatment, the cells were detected for the expression of alkaline phosphatase, Rnh2, stella, fragilis, Tex14, Sry, Hsp90-a, Stra8 and integrin a6, and then, the cells were transplanted into seminiferous tubules of busulfan-treated adult mice. Results Six days after retinoic acid treatment, alkaline phosphatase expressing cells could still be found in embryoid body （EB） derived cells, indicating the existence of retinoic acid-resistant primordial germ cells. When the SSEA-1＋ cells isolated from embryoid bodies were stimulated with retinoic acid for 6 days, some of these cells expressed cell markers of Hsp90-a, Stra8 and integrin a6, resembling the expression profile of spermatogonial stem cells. Forty-five days after cell transplantation, a little amount of GFP-expressing cells attached to the basement membrane of seminiferous tubule and formed small colonies; Three months later, these cells started amplification in the form of cell chains with varied length, and moving towards the lumen of the seminiferous tubules. Five months after the transplantation, multilayered cell mass was found in seminiferous tubules of two, out of four recipient mice. There was no GFP-expressing cells existed in non-cell-transplanted seminiferous tubules. Conclusion In our study, although full-termed spermatogenesis was not observed in all of the recipients, the results did indicate that the embryoid body contains germ like cells, and these cells can survive and initiate amplification in seminiferous tubules of adult mouse.

Genetic algorithm-optimized backpropagation neural network establishes a diagnostic prediction model for diabetic nephropathy:Combined machine learning and experimental validation in mice

Background:Diabetic nephropathy(DN)is the most common complication of type 2 diabetes mellitus and the main cause of end-stage renal disease worldwide.Diagnostic biomarkers may allow early diagnosis and treatment of DN to reduce the prevalence and delay the development of DN.Kidney biopsy is the gold standard for diagnosing DN;however,its invasive character is its primary limitation.The machine learning approach provides a non-invasive and specific criterion for diagnosing DN,although traditional machine learning algorithms need to be improved to enhance diagnostic performance.Methods:We applied high-throughput RNA sequencing to obtain the genes related to DN tubular tissues and normal tubular tissues of mice.Then machine learning algorithms,random forest,LASSO logistic regression,and principal component analysis were used to identify key genes(CES1G,CYP4A14,NDUFA4,ABCC4,ACE).Then,the genetic algorithm-optimized backpropagation neural network(GA-BPNN)was used to improve the DN diagnostic model.Results:The AUC value of the GA-BPNN model in the training dataset was 0.83,and the AUC value of the model in the validation dataset was 0.81,while the AUC values of the SVM model in the training dataset and external validation dataset were 0.756 and 0.650,respectively.Thus,this GA-BPNN gave better values than the traditional SVM model.This diagnosis model may aim for personalized diagnosis and treatment of patients with DN.Immunohistochemical staining further confirmed that the tissue and cell expression of NADH dehydrogenase(ubiquinone)1 alpha subcomplex,4-like 2(NDUFA4L2)in tubular tissue in DN mice were decreased.Conclusion:The GA-BPNN model has better accuracy than the traditional SVM model and may provide an effective tool for diagnosing DN.

Contraceptive effect of Curcuma longa (L.) in male albino rat

Aim: To study the contraceptive effect of the crude extracts of Curcuma longa in male albino rats. Methods: Rats were fed orally with Curcuma longa aqueous and 70 % alcoholic extract for 60 days (500 mg·kg-1· day-1). Results: A reduction in sperm motility and density was observed in both the treated groups. Conclusion: Curcuma longa may have affected the androgen synthesis either by inhibiting the Leydig cell function or the hypo-thalamus pituitary axis and as a result, spermatogenesis is arrested.

Carbon Monoxide Inhibits the Nuclear-cytoplasmic Translocation of HMGB1 in an In Vitro Oxidative Stress Injury Model of Mouse Renal Tubular Epithelial Cells

Carbon monoxide（CO）,as a vital small molecule in signaling pathways,is found to be involved in ischemia-reperfusion injury（IRI） in renal transplantation.CO-releasing molecule-2（CORM-2）,a CO-releasing molecule,is a type of metal carbonyl complexes which can quickly release CO in vivo.In this study,an in vitro oxidative stress injury model was established to examine the effect of CORM-2 pretreatment on the nuclear-cytoplasmic translocation of high mobility group box 1 protein（HMGB1） in mouse primary renal proximal tubular epithelial cells（RPTECs）.Immunofluorescence staining showed that HMGB1 in the medium-and CORM-2-treated groups was predominantly localized in the nucleus of the cells,whereas higher amounts of HMGB1 translocated to the cytoplasm in the H2O2-and inactive CORM-2（i CORM-2）-treated groups.Western blotting of HMGB1 showed that the total amounts of cytoplasmic HMGB1 in the H2O2-treated（0.59±0.27） and i CORM-2-treated（0.57±0.22） groups were markedly higher than those in the medium-treated（0.19±0.05） and CORM-2-treated（0.21±0.10） groups（P〈0.05）.Co-immunoprecipitation showed that the levels of acetylated HMGB1 in the H2O2-treated（642.98±57.25） and i CORM-2-treated（342.11±131.25） groups were markedly increased as compared with the medium-treated（78.72±74.17） and CORM-2-treated（71.42±53.35） groups（P〈0.05）,and no significant difference was observed between the medium-treated and CORM-2-treated groups（P〉0.05）.In conclusion,our study demonstrated that in the in vitro oxidative stress injury model of primary RPTECs,CORM-2 can significantly inhibit the nuclear-cytoplasmic translocation of HMGB1,which is probably associated with the prevention of HMGB1 acetylation.

Detrimental effects of excessive fatty acid secretion on female sperm storage in chickens

Background:Female sperm storage(FSS),the maintenance of sperm inside the female reproductive tract for an extended period of time,is pervasive among organisms with internal fertilization.Because FSS enables asynchronous mating and fertilization,it could be extremely important to reproduction.However,the physiological mechanisms underlying prolonged preservation and maintenance are poorly understood.Here,we used chicken,a typical oviparous animal,to determine the mechanisms ensuring sperm functionality in sperm storage tubules(SSTs).Results:We performed an insemination experiment on over two thousand hens at two periods,and found that the FSS capabilities varied widely among individuals.Except for the differences in the SST density between the two groups with distinct FSS abilities,we quantitatively profiled small-molecule metabolites derived from SST cells,and identified 28 metabolites with differential expression.In particular,high levels of lipids,fatty acids and lipid peroxidation product were observed in hens with low FSS capability.Pathway analysis showed that these differential metabolites were significantly enriched in the biosynthesis of unsaturated fatty acids.Moreover,we detected the total antioxidant capacity and lipid peroxidation level of SSTs,and found that chickens with a lower FSS ability had a significantly higher content of lipid peroxidation end-product,which was 2.4-fold greater than chickens with a higher FSS capability,and no significant difference was found in the total antioxidant capacity between these two groups.Conclusions:Our findings reveal that the long-term storage of sperm and the maintenance of their function in the female reproductive tract require an adequate microenvironment.The superabundance of fatty acids secreted by SST cells had detrimental effects on sperm storage in the female reproductive tract.Lipid peroxidation produces toxic biological substances that may cause irreversible damage to resident spermatozoa,resulting in short-term sperm retention and decreased fertility.Our findings provide new avenues for studying sperm storage and sustaining fertility.

Effect of ligustrum fruit extract on reproduction in experimental diabetic rats

Aim: To study the effect of ligustrum fruit on spermatogenesis and blood gonadal hormones in diabetic rats.Methods: Experimental diabetes was induced in male Wistar rats with streptozotocin. Ligustrum fruit extract wasgiven by gastric gavage at a dose of crude drug 30 g·kg^(-1)·d^(-1) for 110 days. The serum gonadadotropic hormones andtestosterone were determined on d 60 and testicular histology examined on d 110. Results: In the control diabeticrats, the seminiferous tubules were dilated and the spermatogenic cells irregularly arranged. Spermatogenesis was arrest-ed with the number of spermatids highly reduced and spermatozoa not observed. In the treated rats, all types of sper-matogenic cells were practically normal. The serum luteinizing hormone (LH), follicle-stimulating hormone (FSH)and testosterone levels were higher in the treated than in the control rats, but the difference was insignificant. Conclu-sion: In experimental diabetic rats, ligustrum fruit extract protects the damaging effect of experimental diabetes onspermatogenesis. (Asian J Androl 2001 Mar; 3 : 71-73)

Normal and varicocele testis in adolescents

The authors reviewed the results of their research on the structure and composition of normal and varicocele semi-niferous tubules in adolescents. They give new evidences of normal structure of adolescent testis and demonstrate, forthe first time, the ultrastructural and immunohistochemical modifications of the lamina propria and basal lamina in theadolescent varicocele patients, which are similar to those observed in adults, but less severe, and of the adherence junc-tions in seminiferous tubules. They also report the presence of oxidative stress in adolescents limited to testis and notgeneralised as in the adults. These data are well correlated to different clinical studies that support the hypothesis of aprogressive course of varicocele and the need for surgical treatment in adolescent varicocele patients. (Asian J Androl2001 Dec; 3: 259-262)

Effect of lead chloride on spermatogenesis and sperm parameters in mice

Aim: To evaluate the effect of acute lead chloride exposure on testis and sperm parameters in mice. Methods: PbCl2, 74 mg/kg, was daily administered to sexually mature male mice for 3 days and the effects on the testicular histology and ultrastructure as well as the motility and density of spermatozoa in cauda epididymis were observed. An additional group of mice were treated for 1-3 days and were allowed to recover for 32 days to determine the reversibility of lead-induced changes. Results: The testicular weight, seminiferous tubular diameter and sperm counts were significantly decreased following 3 days of PbCl2 treatment, but were unaffected by shorter-term exposures. The changes caused by lead are mostly reversible. Conclusion: Acute lead chloride exposure injures the fertility parameters of male mice and the effects are partially reversible.

Optical coherence tomography of the living human kidney

Acute tubular necrosis(ATN)induced by ischemia is the most common insult to donor kidneysdestined for trarsplantation.ATN results from sweling and subsequent damage to cells lining thelkidney tubules.In this study,we demonstrate the capability of optical coherence tomography(OcT)to image the renal microst ructures of living human donor kidneys and potentially providea measure to det ermine the extent of A TN.We also found that Doppler-based OCT(i.e.,DOCT)reveals renal blood flow dynamics that is another major factor which could relate to post-transplant renal finction.All OCT/DoCT oberva tions were performed in a noninvasive,sterileand timely manner on intact human kidneys both prior to(er vivo)and following(in vivo)theirtransplantation.Our results indicate that this imaging model provides transplant surgeons withan objective visualization of the transplant lidneys prior and immediately post transplantation.