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Vitamin B12-induced spermatogenesis recovery in cimetidine-treated rats： effect on the spermatogonia number and sperm concentration

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Asian Journal of Andrology 2017年第5期19卷 567-572页

作者： Flavia L Beltrame Estela Sasso-Cerri Department of Morphology and Genetics Federal University of Sao Paulo (UNIFESP/EPM) Sao Paulo Brazil Department of Morphology Laboratory f Histology and Embryology Dental School - Sao Paulo State University (UNESP/FOAr) Araraquara Sao Paulo Brazil

The H2-receptor antagonist cimetidine is an antiulcer drug also used for the treatment of cancer due to its antiangiogenic effect. However, this drug has caused structural changes in the seminiferous tubules. Vitamin B12 has been used as a therapeutic agent for the treatment of male infertility. The supplementation of rats with vitamin B12 during cimetidine treatment has recovered the damaged seminiferous tubules, but how this vitamin restores the seminiferous epithelium has not been clarified. In this study, we evaluated whether vitamin B12 improves the number of spermatogonia, spermatocytes, and sperm concentration in cimetidine-treated rats. Adult male rats were treated for 50 days as follows： cimetidine group received 100 mg kg^-1 b.w. of cimetidine, cimetidine-B12 group received cimetidine and 3 μg of vitamin B12-hydroxocobalamin, B12 group received only 3 μg of vitamin, and control group received saline. Sperm concentration was calculated and historesin-embedded testes sections were used for the quantitative analyses of spermatogonia （A; In/B） and spermatocytes. TUNEL method and PCNA immunofluorescence were performed. Cimetidine caused a significant reduction in sperm concentration. TUNEL-positive spermatogonia and spermatocytes were correlated to a significant reduction in the number of these cells. In cimetidine-B12 group, sperm concentration was higher than cimetidine group and a significant increase in the number of spermatogonia （stages Ⅱ-Ⅵ） was correlated to a high incidence of PCNA-immunolabeled spermatogonia and spermatocytes. The results show that the supplementation of rats with vitamin B12 during cimetidine treatment increases sperm concentration and exerts a potential effect in the recovery of spermatogonia and spermatocytes.

关键词： cimetidine fertility spermatogenesis testis vitamin B12

Hippocampus:Molecular,Cellular,and Circuit Features in Anxiety

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Neuroscience Bulletin 2023年第6期39卷 1009-1026页

作者： Hu-Jiang Shi Shuang Wang Xin-Ping Wang Rui-Xin Zhang Li-Juan Zhu Key laboratory of Developmental Genes and Human Diseases MOEDepartment of Histology and EmbryologySchool of MedicineSoutheast UniversityNanjing210009China Shanghai Mental Health Center Shanghai Jiao Tong University School of MedicineShanghai201108China

Anxiety disorders are currently a major psychiatric and social problem,the mechanisms of which have been only partially *** hippocampus serves as a major target of stress mediators and is closely related to anxiety *** so far,its complex anatomy has been a challenge for research on the mechanisms of anxiety *** advances in imaging,virus tracking,and optogenetics/chemogenetics have permitted elucidation of the activity,connectivity,and function of specific cell types within the hippocampus and its connected brain regions,providing mechanistic insights into the elaborate organization of the hippocampal circuitry underlying *** of hippocampal neurotransmitter systems,including glutamatergic,GABAergic,cholinergic,dopaminergic,and serotonergic systems,have contributed to the interpretation of the underlying neural mechanisms of *** and neuroinflammatory factors are also involved in anxiety *** review comprehensively summarizes the hippocampal mechanisms associated with anxiety modulation,based on molecular,cellular,and circuit properties,to provide tailored targets for future anxiety treatment.

关键词： Anxiety Hippocampus Excitatory neurons Interneurons Neural circuit

Glucagon-like peptide-2 modulates the nitrergic neurotransmission in strips from the mouse gastric fundus

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World Journal of Gastroenterology 2017年第40期23卷 7211-7220页

作者： Rachele Garella Eglantina Idrizaj Chiara Traini Roberta Squecco Maria Giuliana Vannucchi Maria Caterina Baccari Department of Experimental and Clinical Medicine Section of Physiology University of Florence Department of Experimental and Clinical Medicine Histology and Embryology Research Unit University of Florence

AIM To investigate whether glucagon-like peptide-2(GLP-2) influences the neurally-induced responses in gastric strips from mice, since no data are available. METHODS For functional experiments, gastric fundal strips were mounted in organ baths containing Krebs-Henseleit solution. Mechanical responses were recorded via forcedisplacement transducers, which were coupled to a polygraph for continuous recording of isometric tension. Electrical field stimulation(EFS) was applied via two platinum wire rings through which the preparationwas threaded. The effects of GLP-2(2 and 20 nmol/L) were evaluated on the neurally-induced contractile and relaxant responses elicited by EFS. Neuronal nitric oxide synthase(n NOS) enzyme was evaluated by *** In the functional experiments, electrical field stimulation(EFS, 4-16 Hz) induced tetrodotoxin(TTX)-sensitive contractile responses, which were reduced in amplitude by GLP-2(P 0.05). The direct smooth muscle response to methacholine was not influenced by GLP-2(P > 0.05). In the presence of guanethidine and carbachol, the addition of GLP-2 to the bath medium evoked TTX-sensitive relaxant responses that were unaffected by L-NNA(P > 0.05). EFS induced a fast NO-mediated relaxation, whose amplitude was enhanced in the presence of the hormone(P < 0.05). Immunohistochemical experiments showed a significant increase(P < 0.05) in n NOS immunoreactivity in the nerve structures after GLP-2 exposure. CONCLUSION The results demonstrate that in gastric fundal strips, GLP-2 influences the amplitude of neurally-induced responses through the modulation of the nitrergic neurotransmission and increases n NOS expression.

关键词： Immunohistochemistry Gastric motility Glucagon-like peptide-2 Neuronal nitric oxide synthase Non-adrenergic non-cholinergic neurotransmission

Characterisation of the bacterial community in expressed prostatic secretions from patients with chronic prostatitis/chronic pelvic pain syndrome and infertile men： a preliminary investigation

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Asian Journal of Andrology 2012年第4期14卷 566-573页

作者： Dong-Sheng Hou Wen-Min Long Jian Shen Li-Ping Zhao Xiao-Yan Pang Chen Xu Department of histology and embryology Shanghai Jiao Tong University School of MedicineShanghai 200025China Shanghai Key laboratory of Reproductive Medicine Shanghai 200025 China National Key laboratory of Microbial Metabolism College of Life Science and BiotechnologyShanghai Jiao Tong UniversityShanghai 200025 China

The expressed prostatic secretions （EPSs） of men with chronic prostatitis/chronic pelvic pain syndrome （CP/CPPS）, infertile men and normal men were subjected to microbiological study. EPSs were collected from the subjects, which included 26 normal men, 11 infertile patients and 51 CP/CPPS patients. DNA was extracted from each specimen, and the V3 regions of the 16S rRNA genes were amplified using universal bacterial primers. The results showed that the EPS 16S rRNA gene-positive rate in the CP/CPPS and infertile patients was much higher than in the normal men, but without any difference among the three patient groups. The denaturing gradient gel electrophoresis （DGGE） method was used to characterize the EPS bacterial community structure of the prostate fluid from patients with CP/CPPS or infertility issues. Principal component analysis （PCA） and partial least squares （PLS） analyses of PCR-DGGE profiles revealed that the EPS bacterial community structure differed among the three groups. Three bands were identified as the key factors responsible for the discrepancy between CP/CPPS patients and infertile patients （P〈O.05）. Two bands were identified as priority factors in the discrepancy of category IliA and category IIIB prostatitis patients （P〈O.05）. According to this research, the ecological balance of the prostate and low urethra tract, when considered as a microenvironment, might play an important role in the maintenance of a healthy male reproductive tract.

关键词： bacteria chronic prostatitis/chronic pelvic pain syndrome denaturing gradient gel electrophoresis prostatitis

Effect of Composite Salvia Injection on Platelet Parameters in Children with Anaphylactoid Purpura

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Chinese Journal of Integrative Medicine 2009年第2期15卷 149-151页

作者：谢雪兰寇素茹许月红李朝英 Department of Pediatrics Affiliated Hospital of Hebei UniversityBaoding Stuff Room of histology Embryology and PediatricsMedical Division of Hebei UniversityBaoding

Objective:To explore the effect of composite salvia injection(CSI) on platelet parameters in children with anaphylactoid purpura(AP) and its clinical ***:One hundred and fifty children with AP were assigned to two groups,80 in Group A and 70 in Group *** were treated,respectively,with conventional therapy only or conventional therapy combined with *** platelet parameters,including blood platelet count(BPC),mean platelet volume(MPV),platelet distribution width(PDW) and plateletcrit(...

关键词： anaphylactoid purpura composite salvia injection platelet parameter

CONSTRUCTION AND IDENTIFICATION OF EUKARYOTIC VECTOR EXPRESSING SIRNA SPECIFIC FOR BACE

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Journal of Pharmaceutical Analysis 2005年第2期17卷 61-65页

作者：董炜疆宫惠琳冯改丰胡海涛王全颖 Department of Anatomy and histology & embryology Medical School of Xi'an Jiaotong University Xi'an 710061 China

Objective To generate eukaryotic expression vector of siRNA specific for β-site APP cleaving enzyme(BACE),and detect the interfering effect to the expression of BACE. Methods To clone the BACE targeting siRNA gene by PCR, the PCR products was inserted into the pUC19/EGFP-U6 plasmid. Then it was sub-cloned into the vector named pLXSN. The resultant plasmid was named pLXSN/EGFP-U6-siBACE, it was packaged into AmphoPack-293 cells by calcium phosphate transfection and collected the virus supernatant. The neuroblastoma cells SK-N-SH was infected with the pLXSN/EGFP-U6-siBACE retroviral vector, immunohistochemistry method was used to detect whether the pLXSN/EGFP-U6-siBACE infection can inhibit the expression of BACE of the neuroblastoma cells. Results The pLXSN/EGFP-U6-siBACE retroviral vector was constructed successfully and the siBACE can inhibit the BACE of the neuroblastma effectively. Conclusion The siRNA can inhibit the expression of the BACE gene, the endogenous production of BACE protein was decreased. It will lay the important foundation for using RNA technology to prevent the Alzheimer's disease.

关键词： siRNA the Alzheimer's disease β-site APP cleaving enzyme

Induced neuronal differentiation of neural stem cells by a combination of cytokines One-step versus two-step methods

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Neural Regeneration Research 2009年第11期4卷 814-821页

作者： Xiaoli Sun Xue Chen Tianyi Zhang Zhenglin Jiang Weiwei Lin Ying Yuan XiaodongWang Department of histology and embryology Medical School of Nantong University Nantong 226001 Jiangsu Province China Affiliated Hospital of Nantong University Nantong 226001 Jiangsu Province China Jiangsu Province Key laboratory of Neuroregeneration Nantong 226001 Jiangsu Province China

BACKGROUND： A combination of basic fibroblast growth factor （bFGF）, platelet-derived growth factor （PDGF）, human heregulin-beta-1, beta-mercaptoethanol retinoic acid and forskolin has been reported to induce the differentiation of rat bone marrow stromal cells into myelinating Schwann-like cells. OBJECTIVE： To investigate the inducing effects of a combination of bFGF, PDGF, human heregulin-beta-1, beta-mercaptoethanol retinoic acid and forskolin on neural stem cell differentiation by one- and two-step methods. DESIGN, TIME AND SETTING： A cytobiology experiment was performed at the Department of histology and embryology, Medical School of Nantong University, and Jiangsu Province Key laboratory of Neuroregeneration, China, between August 2005 and January 2007. MATERIALS： A total of 30 healthy Sprague Dawley rat embryos at gestational days 14-16 were selected, bFGF, PDGF, human heregulin-beta-t, beta-mercaptoethanol, retinoic acid, and forskolin were purchased from Sigma, USA. METHODS： Passage 3 rat neural stem cells were cultured by a one-step method in serum-free medium plus 10 ng/m/bFGF, 5 ng/mL PDGF, 200 ng/mL heregulin-beta-1,35 ng/mL all-trans retinoic acid, and 5 pmol/L forskolin or by a two-step method in serum-free medium plus 35 ng/m/ all-trans retinoic acid for 72 hours, followed by serum-free medium plus 10 ng/mL bFGF, 5 ng/mL PDGF, 200 ng/mL heregulin-beta-t and 5 μmol/L forskolin. The control condition consisted of 10% fetal bovine serum alone or 20 ng/mL bFGF alone. MAIN OUTCOME MEASURES： Differentiated cells were identified by immunocytochemical staining for microtubule associate protein-2 （MAP2） and St 00 protein. Geometric parameters and sodium ion currents of the differentiated cells were measured by image analysis and whole-cell patch-clamp techniques, respectively. RESULTS： Compared with the two-step culture method, neuronal-like cells exhibited longer processes and a similar appearance to mature neurons using the one-step method. The percentage of

关键词： neural stem cells differentiation neuron cytokines whole-cell patch-clamp

Protective effect of low dose of melatonin against cholestatic oxidative stress after common bile duct ligation in rats

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World Journal of Gastroenterology 2005年第13期11卷 1951-1956页

作者： Mukaddes Esrefoglu Mehmet Gül Memet Hanifi Emre Alaattin Polat Mukadder Ayse Selimoglu Department of histology and embryology Department of Physiology Department of Pediatric Gastroenterology

AIM: To investigate the role of oxidative injury and the effect of exogenous melatonin administration on liver damage induced by bile duct ligation (BDL), and second, to evaluate the role of nitric oxide (NO), a free oxygen radical, in oxidative injury. METHODS: Thirty-two Sprague-Dawley rats were assigned to four groups: sham operation (SO), BDL, BDL+melatonin, and BDL+vehicle. Cholestasis was achieved by double ligature of the common bile duct. Melatonin was injected intraperitoneally 500 μg/(kg·d) for 8 d. Hepatic oxidative stress markers were evaluated by changes in the amount of lipid peroxides, measured as malondialdehyde (MDA), and reduced GSH. Total nitrite (NOx) concentrations were determined in hepatic homogenates. Histopathological examination was performed using a histological scoring system. RESULTS: The histopathological changes including portal inflammation, necrosis,apoptosis, focal inflammation and fibrosis were severe in the BDL and BDL+vehicle groups. There were numerous large areas of coagulation necrosis. Histological Activity Index scores of these groups were significantly higher than that of the SO group. Treatment with melatonin reduced these alterations significantly. The degree of necro-inflammation and fibrosis showed significant difference between the BDL and BDL+melatonin groups. BDL was accompanied by a significant increase in MDA and NOx, and a significant decrease in GSH levels. Mean±SE values of MDA, GSH and NOx levels of SO group were 147.47±6.69, 0.88±0.33 μmol/g and 180.70±6.58 nm/g, respectively. The values of BDL group were 200.14±21.30, 0.65±0.02 μmol/g, and 400.46±48.89 nm/g, respectively, whereas the values of BDL+melatonin group were 115.93±6.8,0.74±0.02 μmol/g, and 290.38±32.32 nm/g, respectively. Melatonin treatment was associated with a significant recovery of MDA, GSH and NOx levels. CONCLUSION: We have concluded that oxidative stress is associated with the pathogenesis of cholestatic liver damage and NO contributes to o

关键词： Cholestasis Melatonin Oxidative stress Free radicals Hepatic injury

Increasingβ-hexosaminidase A activity using genetically modified mesenchymal stem cells

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Neural Regeneration Research 2024年第1期19卷 212-219页

作者： Alisa A.Shaimardanova Daria S.Chulpanova Valeriya V.Solovуeva Shaza S.Issa Aysilu I.Mullagulova Angelina A.Titova Yana O.Mukhamedshina Anna V.Timofeeva Alexander M.Aimaletdinov Islam R.Nigmetzyanov Albert A.Rizvanov Institute of Fundamental Medicine and Biology Kazan Federal UniversityKazanRussia Department of histology Cytology and EmbryologyKazan State Medical UniversityKazanRussia

GM2 gangliosidoses are a group of autosomal-recessive lysosomal storage disorde *** diseases result from a deficiency of lysosomal enzymeβ-hexosaminidase A(HexA),which is responsible for GM2 ganglioside *** deficiency causes the accumulation of GM2-gangliosides mainly in the nervous system cells,leading to severe progressive neurodegeneration and *** date,there is no treatment for these ***-mediated gene therapy is considered a promising treatment for GM2 *** study aimed to evaluate the ability of genetically modified mesenchymal stem cells(MSCs-HEXA-HEXB)to restore HexA deficiency in Tay-Sachs disease patient cells,as well as to analyze the functionality and biodistribution of MSCs in *** effectiveness of HexA deficiency cross-correction was shown in mutant MSCs upon intera ction with *** results also showed that the MSCs-HEXA-HEXB express the functionally active HexA enzyme,detectable in vivo,and intravenous injection of the cells does not cause an immune response in *** data suggest that genetically modified mesenchymal stem cells have the potentials to treat GM2 gangliosidoses.

关键词： adeno-associated viral vectors cell therapy cell-mediated gene therapy gene therapy GM2 gangliosidosis Sandhoff disease Tay-Sachs disease β-hexosaminidase