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1.
Syst Biol Reprod Med ; : 1-13, 2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-32040351

RESUMO

The adverse effects of sleep disorders on male fertility are of increased concern. In this study, a rat model of chronic sleep restriction (CSR) was established using the modified multiplatform method. The effects of CSR on the fertility of male rats were evaluated first based on sexual behavior. Serum hormones, including testosterone (T), prolactin (PRL), luteinizing hormone (LH) and follicle-stimulating hormone (FSH), and sperm parameters (concentration, viability, motility, deformation rate) were measured, and testicular histology was analysed by hematoxylin and eosin staining. The transcriptional differences between CSR rats and control rats were detected by RNA sequencing (RNA-Seq), and DNA methylation was then detected by bisulfite sequencing. After the differentialy expressed genes of CSR rats were sequenced and screened, representative up- and down-regulated genes were randomly sampled to verify the sequencing results by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Finally, functional annotations were completed, including gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomic (KEGG) pathway analyses. The results showed that the sexual behavior of CSR rats did not change when compared with control group rats. The sperm concentration, viability and motility of the CSR rats decreased significantly, while the sperm malformation rate increased significantly. In the KEGG pathway analysis database, some specific differentially expressed genes were screened, which are involved in metabolic pathways, inflammation-related pathways, the renin-angiotensin system, as well as others. However, the aforesaid differentially expressed genes in the testes were not related to their DNA methylation status. CSR could significantly reduce the fertility of male rats, and one of its mechanisms occurs by altering gene expression in the testes, which is not related to their  state of  DNA methylation. The results of this study suggest that CSR could cause male infertility by significantly altering the testicular transcriptome.Abbreviations: CSR: chronic sleep restriction; SD: sleep deprivation; RNA-Seq: RNA sequencing; NGS: next generation sequencing; qRT-PCR: real-time quantitative reverse transcription polymerase chain reaction; KEGG: Kyoto encyclopedia of genes and genomic; NO: nitric oxide; INOS: Inducible nitric oxide synthase; Il6: interleukin-6; Tnf: tumour necrosis factor alpha; Hsd11b1: hydroxysteroid 11-beta dehydrogenase 1; Dnmt3a: DNA methyltransferase 3Ax; PSD: paradoxic sleep deprivation; DNMTs: DNA methyltransferases family; REM: rapid eye movement sleep; PGD: preimplantation genetic diagnosis; PGS: preimplantation genetic screening; ECS: expanded carrier screening; T: testosterone; FSH: follicle stimulating hormone; LH: luteinizing hormone; PRL: prolactin; BC group: Blank Control group; MC group: Model Control group; Hist1h2ba: histone cluster 1 H2ba; Lgr4: leucine-rich repeat-containing G protein-coupled receptor 4; Atrn: attractin ; Ogg1: 8-oxoguanine DNA glycosylase; SNVs: single nucleotide variants ; HPG axis: hypothalamic-pituitary-adrenal axis; Star protein: steroid acute regulatory protein; Dmac2l: distal membrane arm assembly complex 2 like; Esr1: estrogen receptor 1; MAPK pathways: mitogen-activated protein kinase pathways; Sos2: SOS Ras/Rho guanine nucleotide exchange factor 2; Jak2: Janus kinase 2; Pik3cb: phosphatidylinositol-4,5-bisphosphate 3-kinase, and catalytic subunit beta; Kras: KRAS proto-oncogene and GTPase; RRBS: reduced representation bisulfite sequencing; DEGs: differently expressed genes; SPF: Specific Pathogen Free; HE: hematoxylin & eosin; DMR: differentially methylated region; GO Analysis: Gene Ontology analysis; SINE: short interspersed nuclear elements; LINE: long interspersed nuclear elements; LTR: long terminal repeats.

2.
BMC Pregnancy Childbirth ; 20(1): 81, 2020 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-32033602

RESUMO

BACKGROUND: The prevalence of both placenta previa and cesarean are on the rise. Multiple adverse outcomes are critically increased when placenta previa is subsequent to prior cesarean. The purpose of the present study is to develop a pre-surgical method for predicting adverse outcomes in pregnancy complicated with both placenta previa and prior cesarean. METHODS: Clinical data was obtained from the medical history system at the First Affiliated Hospital of Sun Yat-sen University from February 2003 to December 2016. All cases with a final diagnosis of "placenta previa/low lying placenta (ICD:O44.001-105)" and "scarred uterus complicated with pregnancy (ICD: O34.200-202)" were collected and reviewed. Hysterectomy was taken as the primary outcome; and blood loss was taken as the secondary outcome. RESULTS: Of 219 pregnant women in the final analysis, 25 received a hysterectomy following delivery, and 48 had blood loss exceeding 1000 ml. Pre-surgical risk factors for hysterectomy are ultrasonic signs of vascular lacunae, central placenta previa, and loss of normal hypoechoic retroplacental zone. A pre-surgical predictive equation referred to as "Hysterectomy Index in Placenta Previa with Prior cesarean (HIPs)" was generated and each risk factor was weighted to create an 8-point scale. This index yielded an area under the curve of 0.972 for the prediction of hysterectomy. CONCLUSIONS: Application of the HIPs score may provide an effective pre-surgical prediction of cesarean hysterectomy in pregnant women complicated with both placenta previa and prior cesarean.

3.
Am J Transl Res ; 11(4): 2580-2589, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31105864

RESUMO

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide and novel therapeutic approaches are urgently required. Anemoside B4 (AB4) is a compound extracted from Pulsatilla chinensis (P. chinensis). Previous studies have indicated that P. chinensis extract P. chinensis saponins has anti-cancer activity. However, the pharmacological effect of AB4 in cancer is largely unknown. In this study, we investigated the anti-cancer efficacy of AB4 in HCC. We used CCK-8 assay and colony formation assay to evaluate the cytotoxicity of AB4 and found that this agent markedly inhibited SMMC7721 cell proliferation. By using a panel of morphological and molecular experiments, we reported that AB4 induced HCC SMMC7721 cell apoptosis and autophagy. Notably, AB4 treatment acts on the Bcl-2-caspase-3 pathway and Beclin-1-LC3-p62 pathway, thereby regulates both apoptosis and autophagy. Finally, we showed that AB4-induced apoptosis and autophagy converges at the PI3K/Akt/mTOR signaling. AB4 treatment inhibits this signaling transduction pathway and leads to HCC cell death. Collectively, our study highlighted the anti-cancer efficacy of AB4 and suggested that AB4 might be a novel way to treat HCC.

4.
Int J Biol Macromol ; 93(Pt A): 136-144, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27246376

RESUMO

In this study, the enzyme-assisted extraction of polysaccharides from Lentinus edodes (LEPs) was optimized by response surface methodology, and a preliminary characterization of the extracted LEPs and their anti-proliferative activities were investigated. An orthogonal assay was constructed to determine the optimal amounts of cellulase, papain and pectinase, which were 15, 20 and 15g/kg, respectively. Then effects of extraction conditions were evaluated and optimized using a Box-Behnken design. The results showed that the highest polysaccharides yield of 15.65% was achieved with an extraction temperature of 54°C, pH 5.0, enzymatic treatment time of 93min and a liquid/material ratio of 29:1mL/g, which correlated well with the predicted yield of 15.58%. Subsequently, the crude LEPs were further purified by DEAE-cellulose and Sephadex-100 chromatography to obtain two fractions, which were designated as LEP-1 and LEP-2 and their monosaccharide compositions were characterized by GC. Fourier-transform infrared spectra demonstrated that LEP-1 and LEP-2 were distinct from each other regarding their chemical structures. In addition, the LEPs exhibited inhibition of cell proliferation on HCT-116 and HeLa cells in vitro. In summary, this study provides an efficient enzyme-assisted extraction for LEPs, which can be used as natural antitumor agents in the pharmaceutical and functional food industries.


Assuntos
Antineoplásicos/isolamento & purificação , Proliferação de Células/efeitos dos fármacos , Polissacarídeos Fúngicos/isolamento & purificação , Cogumelos Shiitake/química , Animais , Antineoplásicos/farmacologia , Celulase/química , Ensaios de Seleção de Medicamentos Antitumorais , Polissacarídeos Fúngicos/farmacologia , Células HCT116 , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Extração Líquido-Líquido , Camundongos , Transplante de Neoplasias , Papaína/química , Poligalacturonase/química , Sarcoma/tratamento farmacológico
5.
Carbohydr Polym ; 131: 322-30, 2015 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-26256191

RESUMO

Response surface methodology was used to optimize the enzyme-assisted extraction parameters for polysaccharides from Tricholoma mongolicum Imai (TMIPs). The orthogonal test design was employed to determine the optimal concentration of three kinds of enzyme (trypsin, pectinase and papain) and the optimal concentrations of trypsin, pectinase and papain were 1.5%, 2.5%, and 2.0% (dry weight % of T. mongolicum Imai powder), respectively. In addition, three variables that remarkably affected the yield of polysaccharides such as extraction temperature, pH and extraction time were studied based on a Box-Behnken design. The results demonstrated that extraction time was the most remarkable factor affecting the TMIPs yield, followed by pH and temperature. Optimal extraction was obtained at 48.4°C, pH 5.4, and extraction time of 132min. Under these optimum conditions, the yield was 24.01%, which is consistent with the predicted value. Furthermore, crude polysaccharides were purified to obtain four fractions. In vitro antioxidant and antiproliferative activities results showed that TMIP-4 had stronger antioxidant and antiproliferative capacity than other fractions.


Assuntos
Antioxidantes/farmacologia , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Tricholoma/química , Análise de Variância , Compostos de Bifenilo/química , Proliferação de Células/efeitos dos fármacos , Depuradores de Radicais Livres/farmacologia , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Radical Hidroxila/química , Picratos/química , Reprodutibilidade dos Testes , Temperatura Ambiente , Fatores de Tempo
6.
Lasers Med Sci ; 30(1): 181-91, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25118661

RESUMO

Photodynamic therapy (PDT) involves the administration and activation of photosensitizing reagents in cancer tissues to induce cytotoxicity. Here we examined the effects of 5-5- (4-N, N-diacetoxylphenyl)-10,15,20- tetraphenylporphyrin (DTPP) -mediated PDT on cell morphology, viability, cell cycle, and cytotoxicity in a murine lung adenocarcinoma cell line. LA795 murine lung adenocarcinoma cell line was used in the study, with cellular uptake of DTPP being quantified by a UV-visible spectrophotometer. The subcellular localization of DTPP was detected by confocal laser scanning microscopy, alteration of cell morphology after PDT was observed by an inverted light microscope, and late-stage apoptosis was examined by terminal dUTP nick end labeling (TUNEL) . The effects of influencing factors on cytotoxicity of PDT in LA795 cells was investigated with varying concentrations of DTPP, energy densities, power densities, and antioxidants by 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Effects of PDT on cell cycle and plasma membrane integrity were studied by flow cytometry analysis. The uptake of DTPP by LA795 cells reached maximum after incubation for 24 h. Confocal laser scanning microscopy showed that DTPP was mainly in the mitochondrion, and slight localization was detected in the lysosomes. Cellular inhibitory effects increased with increased irradiation dose and DTPP concentration, while unactivated DTPP had low toxicity. Flow cytometry analysis revealed that DTPP-PDT-treated cells showed S phase arrest. Cell membrane damage initiation, repair, and irreversible damage were observed at 2, 4, and 5 h after DTPP-PDT , respectively. Together, our results demonstrated cell apoptosis, compromised viability, and cell cycle S phase arrest of LA795 in response to DTPP-PDT , while no effect on the lung cancer cells was observed with irradiation or photosensitizer treatment alone.


Assuntos
Adenocarcinoma/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , Fármacos Fotossensibilizantes/uso terapêutico , Porfirinas/uso terapêutico , Adenocarcinoma/patologia , Adenocarcinoma de Pulmão , Animais , Apoptose/efeitos dos fármacos , Ciclo Celular , Linhagem Celular Tumoral , Membrana Celular/efeitos dos fármacos , Membrana Celular/fisiologia , Forma do Núcleo Celular/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/patologia , Camundongos , Compostos Organofosforados , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/metabolismo , Porfirinas/metabolismo
7.
Pharmacogn Mag ; 9(35): 192-5, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23930000

RESUMO

BACKGROUND: Inula helenium, which belongs to thecomposite family, is an important crude drug in traditional Chinese medicine. MATERIALS AND METHODS: The effects of ethanol concentration, liquid to solid ratio, extraction temperature, and duration of microwave irradiation on the flavonoid extraction yield were studied through a single-factor experiment. An orthogonal array (L9(3(4))) was then constructed to achieve the best extraction conditions. RESULTS: Variance analysis revealed that ethanol concentration significantly affected the extraction yield. The optimal conditions were as follows: ethanol concentration, 50% (v/v); liquid to solid ratio, 15:1; duration of microwave irradiation, 240 s; and extraction temperature, 60°C. CONCLUSION: Under these optimal conditions, the total flavonoid yield was 18.34 ± 0.64 mg/g. The use of a microwave-assisted process dramatically reduced the time needed for extraction of flavonoids from I. helenium.

8.
Acta Crystallogr Sect E Struct Rep Online ; 68(Pt 9): o2714, 2012 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-22969601

RESUMO

In the crystal of the title salt, C(26)H(34)N(2) (2+)·2I(-), the dication lies on a center of inversion that exists along the mid-point of the butyl chain; its five-membered ring is approximately planar (r.m.s. deviation = 0.011 Å). In the crystal, the iodide anion is disordered over two positions in a 1:1 ratio.

9.
Guang Pu Xue Yu Guang Pu Fen Xi ; 29(9): 2551-4, 2009 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-19950673

RESUMO

To examine the difference between early kernels at different ear position in maize and the effect of nitrogen supply rates on kernel development, physiological indices in apical and mid-basal kernel 5-20 days after pollination in maize under different nitrogen supply rates (0, 120, 180 and 240 kg x ha(-1)) were determined with UV-Vis spectrophotometry. The results showed that nitrogen supply obviously increased the contents of soluble sugar, sucrose and starch, and the activities of enzymes related to sucrose inversion and starch synthesis. Twenty days after pollination, the kernel volume, dry weight and the contents of soluble sugar, sucrose and starch in apical kernel under nitrogen supply rate of 180 kg x ha(-1) were higher than those with other treatments. Five to twenty days after pollination, higher activities of acid sucrose invertase (AI), neutral sucrose invertase (NI), sucrose synthase (SS), ADPGase and starch synthase in apical kernel were also obtained under nitrogen supply rate of 180 kg x ha(-1), indicating that the sucrose utilization and starch synthesis were improved, the kernel development was promoted and the kernel abortion was reduced, which resulted in higher yield.


Assuntos
Metabolismo dos Carboidratos , Zea mays , Nitrogênio , Espectrofotometria , Amido , Sacarose , beta-Frutofuranosidase
10.
Yao Xue Xue Bao ; 37(3): 178-80, 2002 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-12579756

RESUMO

AIM: To study the protective effect and mechanism of osthol on learning and memory impairment of mice with acute senile model induced by AlCl3. METHODS: After s.c. AlCl3 60 mg.kg-1 for 7 d and i.p. osthol 15 and 7.5 mg.kg-1 for 12 d, using step-through test and step-down test, the effect of osthol on learning and memory was observed and the glutathione peroxidase (GSH-PX) activities in blood and superoxide dismutase (SOD) activities in plasma and cerebrum were measured. RESULTS: Osthol 15 and 7.5 mg.kg-1 significantly improved the capability of memory and enhanced the activities of GSH-PX and SOD in AlCl3 treated mice. CONCLUSION: Osthol shows protective effect on brain memory impairment of mice in acute senile model induced by AlCl3. Perhaps the mechanism is involved in enhancing the activities of GSH-PX and SOD, clearing away the free radical, protecting the brain neuron from the harm of lipoperoxide.


Assuntos
Envelhecimento/efeitos dos fármacos , Compostos de Alumínio/farmacologia , Cloretos/farmacologia , Cumarínicos/uso terapêutico , Transtornos da Memória/prevenção & controle , Reação de Fase Aguda , Envelhecimento/metabolismo , Cloreto de Alumínio , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Encéfalo/enzimologia , Cnidium/química , Cumarínicos/isolamento & purificação , Cumarínicos/farmacologia , Feminino , Glutationa Peroxidase/sangue , Masculino , Transtornos da Memória/induzido quimicamente , Transtornos da Memória/enzimologia , Camundongos , Plantas Medicinais/química , Distribuição Aleatória , Superóxido Dismutase/metabolismo
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