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1.
Food Funct ; 14(21): 9857-9871, 2023 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-37853817

RESUMO

The protective effects of yak milk (YM) against chronic alcoholic liver injury in rats were investigated in this study. Histologic and biochemical analyses demonstrated that YM consumption ameliorates alcohol-induced liver injury by increasing the liver antioxidant enzyme activity and reducing inflammation. Furthermore, microbiome and metabolomic analyses exploring YM's impact on gut microbiota and metabolism found that YM administration regulates gut microbiota composition. Specifically, there was a decrease in the relative abundance of Helicobacter, Streptococcus, Peptococcus and Tyzzerella, along with an increase in Turisibacter and Intestinimonas. Moreover, Pearson analysis indicated positive correlations between Peptococcus and Tyzzerella with ALT and AST levels, while showing a negative correlation with ADH levels. Furthermore, differential metabolite analysis of fecal samples from the YM group identified significant increases in the taurine (2-Aminoethanesulfonic acid), hypotaurine (2-Aminoethanesulfonic Acid) and isethionic acid levels. Finally, KEGG topology analysis highlighted taurine and hypotaurine metabolism as the primary pathways influenced by YM intervention. Therefore, these findings collectively suggest that YM may protect alcohol-exposed rats against liver injury by modulating oxidative stress, inflammatory response, gut microbiota disorder, and metabolic regulation.


Assuntos
Doença Hepática Crônica Induzida por Substâncias e Drogas , Leite , Ratos , Bovinos , Animais , Doença Hepática Crônica Induzida por Substâncias e Drogas/metabolismo , Fígado/metabolismo , Etanol/toxicidade , Etanol/metabolismo , Taurina/farmacologia , Antioxidantes/farmacologia
2.
Genes (Basel) ; 13(6)2022 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-35741845

RESUMO

Tubb4b (tubulin ß-4b chain) is essential for cell growth and development as a microtubule network protein. Previous studies have shown that TUBB4B affects mouse pronucleus migration, but the gene function has yet to be elucidated. To study TUBB4B-related functions in mouse reproductive development, we designed a single sgRNA in chromosome 2 and generated a knockout spermatogonia cell line of the ß-tubulin isoform Tubb4b by the CRISPR/Cas9 system. Tubb4b-KO spermatogonia recognized abnormal lysosomal membranes and cell morphology defects. Compared to control mouse spermatogonia, the proliferation rate was significantly slower and cycling stagnated in the G1/0 population. Although spermatogonia lacking TUBB4B have abnormal divisions, they are not lethal. We detected the mRNA levels of the cell-regulating cyclins CyclinsD1, CyclinsE, Cdk2, Cdk4, P21, Skp2 and the cell growth factors C/EBP α, C/EBP ß, and G-CSF in the spermatogonia of Tubb4b-KO and found that the expressions of CyclinsD1, Skp2 and cell growth factors were significantly reduced. Further analysis revealed that 675 genes were expressed differently after Tubb4b deletion and were enriched in negative regulation of cell population proliferation (GO:0008285), negative regulation of cell cycle G2/M phase transition (GO:1902750), and positive regulation of cell death (GO: 0010942). We also found that there is a common gene Cdkn1a (P21) in these three GO pathways related to cell proliferation and cell cycle, and both quantitative analysis and transcriptome sequencing results showed that the expression of this gene was up-regulated in Tubb4b knockout cells. This implies that Tubb4b may be involved in the division of spermatogonia with multiple cell cycle regulatory proteins. Overall, these data indicate that Tubb4b has a specific role in regulating spermatogonia proliferation and cell cycle.


Assuntos
Espermatogônias , Tubulina (Proteína) , Animais , Ciclo Celular/genética , Divisão Celular , Proliferação de Células/genética , Masculino , Camundongos , Tubulina (Proteína)/genética
3.
Obes Facts ; 14(5): 559-567, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34515192

RESUMO

INTRODUCTION: The concentration of 25-hydroxycholecalciferol (25OHD3) in the serum of obese people is low and often accompanied by symptoms of low fertility. Therefore, vitamin D is recommended as a potential treatment option. However, after clinical trials, it was found that vitamin D cannot effectively increase the concentration of 25OHD3 in the serum of obese people. How obesity causes low 25OHD3 concentration and low fertility is unclear. METHODS: We analyzed the physiological and pathological changes in obese mice induced by a high-fat diet (HFD) and the changes in mice after supplementing with 25OHD3. RESULTS: The concentration of 25OHD3 in the serum of obese mice induced by HFD was significantly reduced, and these mice showed liver hypertrophy accompanied by abnormal liver injury, testicular hypertrophy, low testosterone levels, high leptin levels, and low sperm motility. The mRNA and protein expression of CYP2R1 of hydroxylated vitamin D3 was significantly reduced; CYP11A1 and CYP11A2, which synthesize testosterone, were significantly reduced. After supplementing with 25OHD3, there was an increase in serum 25OHD3 concentration, testosterone level, and sperm motility, but it cannot improve the degree of obesity, CYP2R1 expression, and liver damage. CONCLUSION: Our research shows that there is a metabolic interference mediated by 25OHD3 and testosterone between obesity and low sperm motility. The results of this study can provide a scientific basis for studying the mechanism of 25OHD3 and hormone regulation and treating obese people with low sperm motility.


Assuntos
Calcifediol , Motilidade dos Espermatozoides , Animais , Fígado , Masculino , Camundongos , Camundongos Obesos , Testosterona
4.
Chem Commun (Camb) ; 57(69): 8632-8635, 2021 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-34369532

RESUMO

Here, a low-defect K2Mn[Fe(CN)6]-reduced graphene oxide (KMF-RGO) composite is fabricated, which demonstrates excellent cycling stability, fast rate capability, and exceptional air stability as a cathode material for potassium-ion batteries (KIBs). This work provides a practical strategy for the synthesis of high-performance K2Mn[Fe(CN)6] cathode materials for KIBs.

5.
Front Vet Sci ; 8: 653413, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33996980

RESUMO

The proteins in the seminal plasma and on the sperm surface play important roles in sperm function and numerous reproductive processes. The cysteine-rich secretory proteins (CRISPs) are enriched biasedly in the male reproductive tract of mammals, and CRISP2 is the sole member of CRISPs produced during spermatogenesis; whereas the role of CRISP2 in fertilization and its association with fertility of boars are still unclear. This study aimed to investigate the relationship between the sperm CRISP2 and boar fertility, and explore its impact sperm fertilizing ability. The levels of CRISP2 protein in sperm were quantified by ELISA; correlation analysis was performed to evaluate the association between CRISP2 protein levels and boar reproductive parameters. Meanwhile, the expression of CRISP2 in boar reproductive organs and sperm, and the effects of CRISP2 on in vitro fertilization (IVF) were examined. The results showed that boars with high sperm levels of CRISP2 had high fertility. The protein levels of CRISP2 in sperm were positively correlated with the litter size (r = 0.412, p = 0.026), the number of live-born piglets (r = 0.421, p = 0.023) and the qualified piglets per litter (r = 0.381, p = 0.042). CRISP2 is specifically expressed in the testis and sperm of adult boars, and its location on sperm changed mainly from the post-acrosomal region to the apical segment of acrosome during capacitation. The cleavage rate was significantly decreased by adding the anti-CRISP2 antibody to the IVF medium, which indicates CRISP2 plays a critical role in fertilization. In conclusion, CRISP2 protein is specifically expressed in the adult testis and sperm and is associated with sperm fertilizing ability and boar fertility. Further mechanistic studies are warranted, in order to fully decipher the role of CRISP2 in the boar reproduction.

6.
Nat Commun ; 12(1): 2167, 2021 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-33846311

RESUMO

Potassium-ion batteries (KIBs) are promising electrochemical energy storage systems because of their low cost and high energy density. However, practical exploitation of KIBs is hampered by the lack of high-performance cathode materials. Here we report a potassium manganese hexacyanoferrate (K2Mn[Fe(CN)6]) material, with a negligible content of defects and water, for efficient high-voltage K-ion storage. When tested in combination with a K metal anode, the K2Mn[Fe(CN)6]-based electrode enables a cell specific energy of 609.7 Wh kg-1 and 80% capacity retention after 7800 cycles. Moreover, a K-ion full-cell consisting of graphite and K2Mn[Fe(CN)6] as anode and cathode active materials, respectively, demonstrates a specific energy of 331.5 Wh kg-1, remarkable rate capability, and negligible capacity decay for 300 cycles. The remarkable electrochemical energy storage performances of the K2Mn[Fe(CN)6] material are attributed to its stable frameworks that benefit from the defect-free structure.

7.
Int J Mol Sci ; 21(16)2020 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-32823735

RESUMO

Tle6 (Transducin-like enhancer of split 6) is a member of the Tle co-repressor superfamily, which is expressed in various tissues of invertebrates and vertebrates and participates in the developmental process. However, the current research has only found that the TLE6 mutation is related to infertility, and the key regulatory mechanism of TLE6 remains to be explored. In this study, we combined Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 and the Tet-on system to construct mouse spermatogonia cell lines that induced TLE6 protein knockout (KO), and studied the effect of Tle6 on mouse spermatogonia proliferation and the cell cycle. The results showed that, after drug induction, the Tle6 gene in mouse spermatogonia was successfully knocked out at the genome and protein levels, and the Tle6 gene knockout efficiency was confirmed to be 87.5% with gene-cloning technology. At the same time, we also found that the mouse spermatogonia proliferated slowly after the Tle6 knockout. Using flow cytometry, we found that the cells did not undergo significant apoptosis, and the number of cells in the S phase decreased. After real-time quantity PCR (qRT-PCR) analysis, we found that the expression of cell-proliferation-related genes, CCAAT enhancer-binding protein α(C/ebp α), granulocyte-colony stimulating factor(G-csf), cyclin-dependent kinases 4(Cdk 4), Cyclin E, proliferating cell nuclear antigen(Pcna), and S-phase kinase-associated protein 2 (Skp2) was significantly reduced, which further affected cell growth. In summary, Tle6 can regulate spermatogonia cell proliferation and the cell cycle and provide a scientific basis for studying the role of TLE6 on spermatogenesis.


Assuntos
Ciclo Celular , Proteínas Correpressoras/genética , Espermatogônias/citologia , Espermatogônias/metabolismo , Animais , Sequência de Bases , Proteína 9 Associada à CRISPR/metabolismo , Sistemas CRISPR-Cas/genética , Proliferação de Células , Proteínas Correpressoras/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Masculino , Camundongos Knockout
8.
Theriogenology ; 156: 107-115, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32698036

RESUMO

Glutathione peroxidases (GPxs) are regarded as important protectors against oxidative stress. Some members of this protein family were reported to play key roles in protecting sperm against oxidative stress. Whether GPx6 a member of the GPx family also plays a role in protection against oxidative stress is not known to date. The objective of the present study was to evaluate the localization and function of glutathione peroxidase 6 (GPx6) in boar accessory sex glands, seminal plasma, and sperm, as well as the effect of GPx6 on vitality and capacitation in boar sperm. qPCR and Western blot analysis demonstrated the presence of GPx6 in testis, epididymis, bulbourethral glands, prostate, seminal vesicle, sperm and seminal plasma. Incubation of sperm with an GPx6 antibody had no significant effect on the viability of boar sperm prior to capacitation. Surprisingly, when capacitated sperm was incubated with the GPx6 antibody for 240 min, sperm vitality was significantly improved. Western blotting showed that in capacitated sperm without prior pretreatment, GPx6 protein content was reduced compared to sperm before capacitation. To further confirm a role for GPx6 in sperm capacitation, we tested sperm acrosome reaction by ACR.2 and FITC-PSA. The results showed that treatment of sperm with the GPx6 antibody significantly increased sperm capacitation and acrosome reaction. Furthermore, we examined the concentration of cAMP in sperm after capacitation. ELISA demonstrated that the cAMP concentration in the sperm exposed to the GPx6 antibody was significantly higher than that of the control group. In addition, the exposure of sperm to the GPx6 antibody significantly increased the concentration of H2O2, while the expression of SOD3 and CAT were decreased. Based on these observations we would like to postulate that in the boar reproductive tract the GPx6 protein becomes attached to the sperm head preventing the sperm to undergo premature capacitation by affecting components of the antioxidant pathway. How GPx6 expression following ejaculation becomes suppressed to allow sperm capacitation to take place needs further investigation.


Assuntos
Reação Acrossômica , Peróxido de Hidrogênio , Acrossomo , Animais , Masculino , Sêmen , Capacitação Espermática , Espermatozoides , Suínos
9.
Anim Sci J ; 91(1): e13391, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32558027

RESUMO

One hundred Yorkshire × Landrace sows were randomly assigned to one of two dietary treatments (diet ND: 6,000 IU vitamin D3 /d feed; diet 25-D: 200 µg/day 25OHD3 feed). The experiment began on d 90 of gestation and continued until weaning on day 21 of lactation. In sows that received 25OHD3 , the growth rate of the piglets before weaning was significantly accelerated (0.266 kg/day, p < .05). Sow serum was collected after weaning, and those in the 25OHD3 group were found to have significantly higher serum calcium (CA) and phosphorus (PI) levels (p < .05). Interestingly, the oestrus cycle of sows fed 25OHD3 was significantly shortened (p < .05), the oestrus time was concentrated on the fifth day after weaning, and the piglets were born with a higher degree of uniformity (p < .05). Colostrum was collected on the day of delivery, and the colostrum of sows fed 25OHD3 contained higher milk fat content than the control group (p < .05). 25OHD3 supplementation increased the mRNA and protein expression of INSIG1 and SREBP1, which regulate milk fat synthesis, in the mammary gland of lactating sows (p < .05). In conclusion, 25OHD3 supplementation in maternal diets improved reproductive performance, milk fat content and the mRNA and protein levels of genes regulating milk fat synthesis in lactating sows.


Assuntos
25-Hidroxivitamina D 2/administração & dosagem , 25-Hidroxivitamina D 2/farmacologia , Fenômenos Fisiológicos da Nutrição Animal/genética , Fenômenos Fisiológicos da Nutrição Animal/fisiologia , Dieta/veterinária , Suplementos Nutricionais , Expressão Gênica/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Lactação/genética , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Reprodução/efeitos dos fármacos , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Suínos/genética , Suínos/fisiologia , Animais , Cálcio/metabolismo , Estro/efeitos dos fármacos , Feminino , Glicolipídeos/metabolismo , Glicoproteínas/metabolismo , Lactação/fisiologia , Gotículas Lipídicas , Fósforo/metabolismo , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
10.
Cells ; 9(1)2020 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-31936362

RESUMO

Granulosa cells (GCs) are the key components of ovarian follicles for regulating oocyte maturation. Previous established GC lines have allowed prolonged proliferation, but lost some physiological features owing to long-term immortalization. This study was to establish an induced immortal porcine GC line with reversible proliferation status by the tetracycline inducible (Tet-on) 3G system. Our conditional immortal porcine GCs (CIPGCs) line steadily propagated for at least six months and displayed primary GC morphology when cultured in the presence of 50 ng/mL doxycycline [Dox (+)]. Upon Dox withdrawal [Dox (-)], Large T-antigen expression, reflected by mCherry fluorescence, gradually became undetectable within 48 h, accompanied by less proliferation and size increase. The levels of estradiol and progesterone, and the expression of genes associated with steroid production, such as CYP11A1 (cytochrome P450 family 11), 3ß-HSD (3ß-hydroxysteroid dehydrogenase), StAR (steroidogenic acute regulatory protein), and CYP19A1 (cytochrome P450 family 19 subfamily a member 1), were all significantly higher in the Dox (-) group than Dox (+) group. The CIPGCs could switch into a proliferative state upon Dox induction. Interestingly, the expression of StAR and CYP19A1 in the CIPGCs (-Dox) was significantly increased by adding porcine follicular fluid (PFF) to mimic an ovary follicle environment. Moreover, PFF priming the CIPGCs in Dox (-) group resulted in similar estradiol production as that of primary GC, and enabled this cell line to respond to gonadotrophins in estradiol production. Collectively, we have established an inducible immortal porcine GC line, which offers a unique and valuable model for future research on the regulation of ovarian functions.


Assuntos
Proliferação de Células , Doxorrubicina/farmacologia , Estradiol/metabolismo , Células da Granulosa/citologia , Progesterona/metabolismo , Animais , Antibióticos Antineoplásicos/farmacologia , Aromatase/genética , Aromatase/metabolismo , Ciclo Celular , Feminino , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/metabolismo , Suínos
11.
Mol Med Rep ; 20(5): 4013-4022, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31545448

RESUMO

Testicular seminoma is one of the most common tumours in the field of urology, and its aetiology is still unclear. The aim of the present study was to identify the factors responsible for the development of testicular cancer and to investigate whether mutations in these genes were primarily congenital or acquired. To identify the key genes and miRNAs linked to testicular seminoma, as well as their potential molecular mechanisms, the GSE15220, GSE1818 and GSE59520 microarray datasets were analysed. A total of 5,195 and 1,163 differentially expressed genes (DEGs) were identified after analysing the GSE15220 and GSE1818 datasets, respectively. Among them, 287 genes were common between the two datasets. Of these, 110 were upregulated and 177 were downregulated. Five differentially expressed microRNAs (miRs; DEMs) that were downregulated in seminoma were identified after analysing the GSE59520 dataset. Following protein­protein interaction network and Gene Ontology analysis, the five nodes with the highest degrees were screened as hub genes. Among them, the high expression of hub genes, such as protein tyrosine phosphatase receptor type C (PTPRC), was associated with worse overall survival. We also predicted the potential target genes of the DEMs. DNA topoisomerase II α (TOP2A), marker of proliferation Ki­67 (MKI67), PTPRC and ubiquitin conjugating enzyme E2 C were associated with the PI3K/AKT and Wnt/ß­catenin signalling pathways. In addition, hsa­miR­650 and hsa­miR­665 were associated with the PI3K/AKT and Wnt/ß­catenin signalling pathways. Additionally, TOP2A and MKI67 were strongly associated with the target genes hsa­miR­650 and hsa­miR­665, respectively. We proposed that the hub genes reported in the present study may have a certain impact on cellular proliferation and migration in testicular seminoma. The roles of these hub genes in seminoma may provide novel insight to improve the diagnosis and treatment of patients with seminoma.


Assuntos
Biomarcadores Tumorais , Biologia Computacional , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Seminoma/genética , Neoplasias Testiculares/genética , Bases de Dados Genéticas , Perfilação da Expressão Gênica , Ontologia Genética , Humanos , Masculino , Mapeamento de Interação de Proteínas , Mapas de Interação de Proteínas , Transcriptoma
12.
ACS Appl Mater Interfaces ; 11(25): 22449-22456, 2019 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-31150200

RESUMO

Recently, nonaqueous potassium-ion batteries (KIBs) are attracting because of increasing interest due to the abundance of potassium resources, but the systematic study about the effects of electrolyte's salt on the electrochemical performance of electrode materials is still insufficient. Here, it is shown that the capacity retention and Coulombic efficiency of commercial micrometric MoS2 can be remarkably improved by simply using potassium bis(fluorosulfonyl)imide (KFSI) over potassium hexafluorophosphate (KPF6) dissolved in ethylene carbonate/diethyl carbonate as the electrolyte. By constructing various cell configurations, it is discovered that the degradation of MoS2||K half-cells in KPF6-containing electrolyte originates from the failure of the MoS2 electrode. The solid electrolyte interphase (SEI) layer formed on MoS2 during cycling was systematically investigated by using a series of characterizations. It is found that a stable, protective, and KF-rich SEI layer is formed on MoS2 in the KFSI-containing electrolyte, while an unstable, KF-deficient, and organic species-rich SEI layer is formed in the KPF6-containing electrolyte. Finally, the origins of such differences are discussed, which will provide new insights into further exploration of novel electrolytes for KIBs.

13.
Mol Med Rep ; 19(1): 155-164, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30483791

RESUMO

Pronuclear migration, which is the initial stage of embryonic development and the marker of zygote formation, is a crucial process during mammalian preimplantation embryonic development. Recent studies have revealed that long non­coding RNAs (lncRNAs) serve an important role in early embryonic development. However, the functional regulation of lncRNAs in this process has yet to be elucidated, largely due to the difficulty of assessing gene expression alterations during the very short time in which pronuclear migration occurs. It has previously been reported that migration of the pronucleus of a zygote can be obstructed by simulated microgravity. To investigate pronuclear migration in mice, a rotary cell culture system was employed, which generates simulated microgravity, in order to interfere with murine pronuclear migration. Subsequently, lncRNA sequencing was performed to investigate the mechanism underlying this process. In the present study, a comprehensive analysis of lncRNA profile during the mouse pronuclear stage was conducted, in which 3,307 lncRNAs were identified based on single­cell RNA sequencing data. Furthermore, 52 lncRNAs were identified that were significantly differentially expressed. Subsequently, 10 lncRNAs were selected for validation by reverse transcription­quantitative polymerase chain reaction, in which the same relative expression pattern was observed. The results revealed that 12 lncRNAs (lnc006745, lnc007956, lnc013100, lnc013782, lnc017097, lnc019869, lnc025838, lnc027046, lnc005454, lnc007956, lnc019410 and lnc019607), with tubulin ß 4B class IVb or actinin α 4 as target genes, may be associated with the expression of microtubule and microfilament proteins. Binding association was confirmed using a dual­luciferase reporter assay. Finally, Gene Ontology analysis revealed that the target genes of the differentially expressed lncRNAs participated in cellular processes associated with protein transport, binding, catalytic activity, membrane­bounded organelle, protein complex and the cortical cytoskeleton. These findings suggested that these lncRNAs may be associated with migration of the mouse pronucleus.


Assuntos
Regulação da Expressão Gênica/genética , Redes Reguladoras de Genes/genética , RNA Longo não Codificante/genética , Animais , Desenvolvimento Embrionário/genética , Feminino , Perfilação da Expressão Gênica/métodos , Ontologia Genética , Camundongos , Camundongos Endogâmicos C57BL , Ausência de Peso , Simulação de Ausência de Peso/métodos
14.
Stem Cell Res Ther ; 9(1): 200, 2018 07 18.
Artigo em Inglês | MEDLINE | ID: mdl-30021628

RESUMO

BACKGROUND: Male germline stem cells (mGSCs) offer great promise in regenerative medicine and animal breeding due to their capacity to maintain self-renewal and to transmit genetic information to the next generation following spermatogenesis. Human testis-derived embryonic stem cell-like cells have been shown to possess potential of mesenchymal progenitors, but there remains confusion about the characteristics and origin of porcine testis-derived stem cells. METHODS: Porcine testis-derived stem cells were obtained from primary testicular cultures of 5-day old piglets, and selectively expanded using culture conditions for long-term culture and induction differentiation. The stem cell properties of porcine testis-derived stem cells were subsequently assessed by determining the expression of pluripotency-associated markers, alkaline phosphatase (AP) activity, and capacity for sperm and multilineage differentiation in vitro. The gene expression profile was compared via microarray analysis. RESULTS: We identified two different types of testis-derived stem cells (termed as C1 and C2 here) during porcine testicular cell culture. The gene expression microarray analysis showed that the transcriptome profile of C1 and C2 differed significantly from each other. The C1 appeared to be morphologically similar to the previously described mouse mGSCs, expressed pluripotency- and germ cell-associated markers, maintained the paternal imprinted pattern of H19, displayed alkaline phosphatase activity, and could differentiate into sperm. Together, these data suggest that C1 represent the porcine mGSC population. Conversely, the C2 appeared similar to the previously described porcine mGSCs with three-dimensional morphology, abundantly expressed Leydig cell lineage and mesenchymal cell-specific markers, and could differentiate into testosterone-producing Leydig cells, suggesting that they are progenitor Leydig cells (PLCs). CONCLUSION: Collectively, we have established the expected characteristics and markers of authentic porcine mGSCs (C1). We found for the first time that, the C2, equivalent to previously claimed porcine mGSCs, are actually progenitor Leydig cells (PLCs). These findings provide new insights into the discrepancies among previous reports and future identification and analyses of testis-derived stem cells.


Assuntos
Células Intersticiais do Testículo/metabolismo , Testículo/metabolismo , Células-Tronco Germinativas Adultas , Animais , Diferenciação Celular , Humanos , Masculino , Suínos
15.
Int J Mol Med ; 38(2): 423-32, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27353491

RESUMO

Mouse spermatogonial stem cells (mSSCs) may be reprogrammed to become pluripotent stem cells under in vitro culture conditions, due to epigenetic modifications, which are closely associated with the expression of transcription factors and epigenetic factors. Thus, this study was conducted to compare the gene expression of transcription factors and epigenetic factors in mSSCs and mouse embryonic stem cells (mESCs). Firstly, the freshly isolated mSSCs [mSSCs (f)] were enriched by magnetic-activated cell sorting with Thy1.2 (CD90.2) microbeads, and the typical morphological characteristics were maintained under in vitro culture conditions for over 5 months to form long-term propagated mSSCs [mSSCs (l)]. These mSSCs (l) expressed pluripotency­associated genes and were induced to differentiate into sperm. Our findings indicated that the mSSCs (l) expressed high levels of the transcription factors, Lin28 and Prmt5, and the epigenetic factors, Tet3, Parp1, Max, Tert and Trf1, in comparison with the mESCs, with the levels of Prmt5, Tet3, Parp1 and Tert significantly higher than those in the mESCs. There was no significant difference in Kdm2b expression between mSSCs (l) and mESCs. Furthermore, the gene expression of N-Myc, Dppa2, Tbx3, Nr5a2, Prmt5, Tet3, Parp1, Max, Tert and Trf1 in the mSSCs (l) was markedly higher in comparison to that in the mSSCs (f). Collectively, our results suggest that the mSSCs and the mESCs displayed differential gene expression profiles, and the mSSCs possessed the potential to acquire pluripotency based on the high expression of transcription factors and epigenetic factors. These data may provide novel insights into the reprogramming mechanism of mSSCs.


Assuntos
Perfilação da Expressão Gênica , Células-Tronco Embrionárias Murinas/metabolismo , Espermatogônias/citologia , Animais , Diferenciação Celular/genética , Separação Celular , Epigênese Genética , Masculino , Camundongos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Transcrição/metabolismo
16.
Yi Chuan ; 36(8): 747-55, 2014 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-25143272

RESUMO

The process of mammalian fertilization involves a series of sperm functional activities, including the sperm transportation, hyperactivation and capacitation, acrosome reaction and sperm-egg fusion, etc. The semen proteins play indispensable roles in these processes, and they are closely associated with the fecundity of males. So these proteins can be biomarkers to evaluate the fertilization capacity of mammalian semen. In this review, we mainly introduce some semen proteins relevant to spermatozoa functions and illustrate their important regulatory roles on the fertilization processes, involving spermatozoa motility, capacitation, acrosome reaction, penetrating the zona pellucida and sperm-egg fusion, etc. Moreover, their potential applications in the evaluation of heredity in livestock are also summarized.


Assuntos
Fertilização , Proteínas/metabolismo , Espermatozoides/metabolismo , Animais , Humanos , Masculino , Espermatozoides/citologia , Espermatozoides/fisiologia
17.
Zhonghua Yi Xue Za Zhi ; 94(11): 852-4, 2014 Mar 25.
Artigo em Chinês | MEDLINE | ID: mdl-24854755

RESUMO

OBJECTIVE: To explore the feasibility, advantages and clinical value of gasless abdominal-wall lifting laparoscopic myomectomy with 5 mm laparoscope and 2 abdominal holes (1.5-hole-gasless-laparoscopic myomectomy). METHODS: A total of 90 cases of uterine fibroids were randomly divided into 2 groups. Lifting gasless group (n = 46) underwent gasless abdominal-wall lifting laparoscopic myomectomy with 5 mm laparoscope and 2 abdominal holes, and pneumoperitoneum group (n = 44) pneumoperitoneum laparoscopic myomectomy. The operative duration, blood loss volume, average time of single-myoma-removal, intestinal function recovery and hospital stay of both groups were compared. RESULTS: The operative duration, blood loss volume and average time of single-myoma-removal of lifting gasless group were respectively significantly less than those of pneumoperitoneum group (P < 0.01) . The postoperative intestinal function recovery and postoperative hospital stay had no significant difference between two groups (P > 0.05). Three cases of pneumoperitoneum group were converted successfully into myomectomy with traditional 3-hole gasless abdominal wall lifting laparoscopy because of large fibroids in uterine isthmus. A total of 12 newly discovered myomas, not pre-detected ultrasonically, were removed in 10 cases of lifting gasless group. CONCLUSION: 1.5-hole-gasless-laparoscopic myomectomy, like traditional gasless laparoscopy, may avoid the complications of laparoscopic CO2 pneumoperitoneum. The smaller laparoscope-hole and sole operating hole make this maneuver a safe, easy and mini-invasive procedure. It is more suitable for clinical application and popularity in primary care.


Assuntos
Laparoscopia/métodos , Leiomioma/cirurgia , Neoplasias Uterinas/cirurgia , Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Pneumoperitônio Artificial/métodos , Resultado do Tratamento
18.
Zhonghua Fu Chan Ke Za Zhi ; 40(10): 656-8, 2005 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-16277892

RESUMO

OBJECTIVE: To compare the clinical characteristics of transvaginal hysterectomy (TVH) and total laparoscopic hysterectomy (TLH). METHOD: Clinical data about 301 cases who received TVH and TLH were collected and the hospital stay days, medical expenses, diagnoses, operation and recovery status were compared between TVH and TLH groups. RESULTS: The ratio of cervical atypical hyperplasia (9.64%), multipara (96.45%) in TVH was higher than that in TLH (2.88%, 89.42%). The ratio of adenoma (29.44%), adnexal disease (4.55%), pelvic endometriosis (4.06%), history of cesarean section (7.11%) in TVH were lower than that in TLH (43.27%, 31.73%, 12.50%, 24.04%). The operation time (76 +/- 28) minutes, bleeding during operation (170 +/- 125) ml, additional operations (5.08%), pelvic adhesion (4.57%), loosening of pelvic adhesion (0.51%), the diameter of the largest myoma or adenoma (49 +/- 17) mm, expenses for operation and hospitalization (1073 +/- 203) yuan in TVH were lower than those in TLH, which were (139 +/- 52) minutes, (206 +/- 153) ml, 36.54%, 41.35%, 17.31%, (57 +/- 22) mm, (1526 +/- 676) yuan respectively. The differences were significant (all P < 0.05). There was no difference of the uterine weight, complication and length of hospitalization duration between the two kinds of operation. CONCLUSIONS: TVH is recommended in cases of few pelvic adhesion, or adnexal disease, cervical disease and of multipara. The uterine weight is not a decisive factor.


Assuntos
Histerectomia Vaginal/métodos , Histerectomia/métodos , Fatores Etários , Peso Corporal , Feminino , Hemorragia/etiologia , Humanos , Histerectomia/efeitos adversos , Histerectomia/economia , Histerectomia Vaginal/efeitos adversos , Histerectomia Vaginal/economia , Complicações Intraoperatórias/etiologia , Laparoscopia , Tempo de Internação , Resultado do Tratamento
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