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OBJECTIVE: To explore the preventive and therapeutic effects of Icariside â ¡ (ICAâ ¡) on radiation injury-induced ED (Ri-ED) in rats. METHODS: Twenty-four 10-week-old male SD rats received exposure of the prostate to single X-ray irradiation of 20 Gy, and were randomly equally divided into an Ri-ED group (6 survived and 6 died) and a treatment group treated with ICAâ ¡ at 4.5 mg/kg/d (9 survived and 3 died). Another 6 SD rats were taken as negative controls. After 4 weeks of continuous intragastric administration and 2 weeks of drug elution, the erectile function of the rats was evaluated by measurement of the maximum intracavernous pressure / mean arterial pressure (ICPmax/MAP), and the penile tissues were subjected to immunofluorescence staining, immunohistochemistry, Masson's trichrome staining, Western blot and detection of oxidative stress indicators. RESULTS: Compared with the negative controls, the rats in the Ri-ED group showed significant decreases in ICPmax/MAP (0.76 ± 0.09 vs 0.42 ± 0.06, P < 0.01), the number of nNOS-positive nerve fibers in the corpus cavernosum (10.17 ± 2.64 vs 3.17 ± 1.72, P < 0.01), the content of endothelial cells (1.39 ± 0.30 vs 0.35 ± 0.12, P < 0.01), the expressions of nNOS (0.42 ± 0.08 vs 0.08 ± 0.01, P < 0.01) and eNOS (0.99 ± 0.24 vs 0.12 ± 0.08, P < 0.01) and the activity of superoxide dismutase (SOD) (ï¼»343.73 ± 58.57ï¼½ vs ï¼»153.50 ± 34.06ï¼½ U/mg prot, P < 0.01), but an increase in the malondialdehyde (MDA) level (ï¼»1.80 ± 0.31ï¼½ vs ï¼»3.25 ± 0.21ï¼½ nmol/mg prot, P < 0.01). In comparison with the Ri-ED group, the animals treated with ICAâ ¡ exhibited remarkably increased ICP/MAP (0.42 ± 0.06 vs 0.66 ± 0.07, P < 0.01), number of nNOS-positive nerve fibers (3.17 ± 1.72 vs 7.33 ± 1.75, P < 0.05), content of endothelial cells (0.35 ± 0.12 vs 1.07 ± 0.36, P < 0.01), and expressions of nNOS (0.08 ± 0.01 vs 0.16 ± 0.05, P < 0.05) and eNOS (0.12 ± 0.08 vs 0.86 ± 0.30, P < 0.01) in the corpus cavernosum, but a decreased level of MDA (ï¼»3.25 ± 0.21ï¼½ vs ï¼»2.17 ± 0.55ï¼½ nmol/mg prot, P < 0.05). In addition, ICAâ ¡ effectively reduced radiation injury-induced mortality of the rats. CONCLUSION: Icarisideâ ¡ can significantly improve ED and pathological changes and reduce mortality caused by radiation injury in rats, which may be related to its ability of improving radiation-induced oxidative stress.
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Disfunción Eréctil , Traumatismos por Radiación , Humanos , Ratas , Masculino , Animales , Disfunción Eréctil/tratamiento farmacológico , Disfunción Eréctil/etiología , Disfunción Eréctil/prevención & control , Células Endoteliales , Ratas Sprague-Dawley , Erección Peniana/fisiología , Pene/patologíaRESUMEN
OBJECTIVE: To investigate the effect and safety of low-intensity pulsed ultrasound (LIPUS) versus low-energy shock wave (LESW) in the treatment of neurogenic penile ED in male SD rats. METHODS: Twenty-four 12-week-old male SD rats were randomly divided into four groups of an equal number: sham operation, bilateral cavernous nerves injury (BCNI), LIPUS (300 mW /cm2, 3 times a week for 2 weeks), and LESW (300 strokes once, 3 times a week for 2 weeks). At 28 days after intervention, the erectile function of the rats was assessed by comparing the ratio of maximum intracavernous pressure to mean arterial pressure (ICPmax/MAP), and the histopathological changes in the corpus cavernosum of the penis were observed by immunohistochemistry, immunofluorescence staining, and Masson trichromatic staining. RESULTS: After treatment, the LIPUS and LESW groups, compared with the BCNI group, showed significantly increased ICPmax/MAP ratio (0.56 ± 0.13 and 0.55 ± 0.10 versus 0.35 ± 0.14, P = 0.017 and P = 0.013), improved smooth muscle/collagen value (0.08 ± 0.01 and 0.08 ± 0.02 versus 0.06 ± 0.02, P = 0.017 and P = 0.019), and elevated proportion of smooth muscle to cavernosum (0.20 ± 0.05 and 0.21 ± 0.03 versus 0.15 ± 0.02, P = 0.046 and P = 0.020), with no statistically significant difference between the LIPUS and LESW groups. No obvious adverse reactions were observed in the LIPUS or LESW group. CONCLUSIONS: Both LIPUS and LESW can effectively improve penile erectile function and repair histopathological injury in the animal model of neurogenic ED.
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Disfunción Eréctil , Animales , Disfunción Eréctil/terapia , Humanos , Masculino , Pene , Ratas , Ratas Sprague-Dawley , Ondas UltrasónicasRESUMEN
In this study, we developed a novel liquid fermentation medium of Cordyceps militaris using pupa powder and wheat bran as nitrogen resources instead of the traditionally used peptone. This process not only reduced the cost by approximately 50%, but increased production by over 30%. Then, we explored a method to extract and purify cordycepin by combining hydrothermal reflux extraction with macroporous resin adsorption, which is inexpensive and suitable for the industrial production. The optimum conditions for hydrothermal reflux were extracting three times at 95 °C with 1:10 sample-to-water ratio, and the cordycepin purity with macroporous resin HPD-100 reached 95.23%.[Formula: see text].
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Cordyceps , Desoxiadenosinas , Fermentación , Estructura MolecularRESUMEN
OBJECTIVE: To construct the eukaryotic express vector containing apoptosis-inducing factor (AIF) gene and to study its expression in A549 cells. METHODS: According to the GenBank AIF mRNA sequence, specific primers to amplify AIF gene from lung carcinoma cell line A549 by RT-PCR was designed. The amplified AIF gene fragment was cloned into plasmid pUC-T by TA cloning, then double enzyme digestion and DNA sequencing were used to identifying the positive recombinant AIF-pUC-T. The target fragment was retrieved and cloned into the eukaryotic express vector pcDNA3.1(+). The positive recombinant AIF-pcDNA3.1(+) was transfected into A549 cells, and expression of AIF gene was verified by RT-PCR and Western blot. RESULTS: AIF target gene was successfully amplified and cloned into the pUC-T. The target fragment was retrieved and cloned into the eukaryotic express vector pcDNA3.1(+), and it was completely coincided with the AIF sequence in GenBank suggested by cells transfected with AIF-pcDNA3. 1(+) was much higher than that of control cells which was not transfected with AIF-pcDNA3.1(+). CONCLUSION: The AIF eukaryotic expression vector AIF-pcDNA3.1(+) is successfully constructed in A549 cells and it could be experimental foundations for further study of AIF gene.
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Factor Inductor de la Apoptosis/biosíntesis , Vectores Genéticos/genética , Transfección , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Factor Inductor de la Apoptosis/genética , Secuencia de Bases , Clonación Molecular , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Datos de Secuencia Molecular , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Células Tumorales CultivadasRESUMEN
N-doped mesoporous carbon spheres (NHMC@mSiO2 ) encapsulated in silica shells were prepared by emulsion polymerization and domain-limited carbonization using ethylenediamine as the nitrogen source, and Ru-Ni alloy catalysts were prepared for the hydrogenation of α-pinene in the aqueous phase. The internal cavities of this nanomaterial are lipophilic, enhancing mass transfer and enrichment of the reactants, and the hydrophilic silica shell enhances the dispersion of the catalyst in water. N-doping allows more catalytically active metal particles to be anchored to the amphiphilic carrier, enhancing its catalytic activity and stability. In addition, a synergistic effect between Ru and Ni significantly enhances the catalytic activity. The factors influencing the hydrogenation of α-pinene were investigated, and the optimum reaction conditions were determined to be as follows: 100 °C, 1.0â MPa H2 , 3â h. The high stability and recyclability of the Ru-Ni alloy catalyst were demonstrated through cycling experiments.
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This work aims to investigate the therapeutic effect of ursolic acid (UA) plus insulin (In) on diabetic nephropathy (DN) in streptozotocin (STZ)-induced T1DM rats. The experimental groups and operational details are as follows: A total of thirty-two SD rats were divided into four groups: the DN model group (DN, n = 8), DN + In treatment group (DN + In, n = 8), DN + In + UA administration group (DN + In + UA, n = 8), and negative control group (control, n = 8). After 8 weeks, changes in renal function indices and pathological damage were assessed. Additionally, oxidative stress-, apoptosis-, and fibrosis-related proteins in kidney tissue were measured. Compared with the control group, the vehicle group showed higher levels of creatine, blood urea nitrogen, urinary protein, apoptosis, and lipid peroxidation; lower superoxide dismutase levels; more severe levels of pathological kidney damage and renal fibrosis; and a deepened degree of EMT and EndMT. Better outcomes were achieved with the combined treatment than with insulin-only treatment. The improvement of TGF-ß1, phosphorylated p38 MAPK, FGFR1, SIRT3 and DPP-4 expression levels in renal tissues after combination therapy was greater than that after insulin-only treatment. This study shows that the combination of insulin and UA significantly improved the pathological changes in the renal tissue of T1DM rats, and the underlying mechanism may be related to improving apoptosis and oxidative stress by regulating p38 MAPK, SIRT3, DPP-4 and FGFR1 levels, thereby blocking TGF-ß signaling pathway activation and inhibiting EMT and EndMT processes.
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OBJECTIVE: To explore the effect of DNA polymerase beta (pol beta) expression level on biological characteristics of mouse embryonic fibroblast (MEF) and the cellular response to DNA damage induced by potassium dichromate. METHODS: pol beta wild-type cells (pol beta +/+), pol beta null cells (pol beta -/-) and pol beta overexpressed cells (pol beta oe) were applied as a model system. The growth curve of cells was plotted by MTT assay; the doubling time of cells was detected by double time experiment; the spontaneous mutation frequency was determined by HGPRT gene mutation method and single cell gel electrophoresis assay (SCGE) was employed to observe the DNA damage either happened spontaneously or induced by potassium dichromate. RESULTS: Growth characteristic and doubling time of the three kinds of cells were similar and no obvious differences were found on spontaneous DNA damage and mutations frequency among them (P > 0.05). Potassium dichromate increased comet rate and tail length in the three kinds of cells in a concentration dependent way. DNA damage of pol beta -/- cells at the same dosage were more serious than the other cells both in comet rate and tail length (P < 0.05). pol beta oe cells demonstrated more resistant to DNA damage obviously than the others. CONCLUSION: The expression level of pol beta has no significant effect on the biological characteristic and spontaneous mutation frequency of MEF. pol beta knock out cells is more sensitive to DNA damage induced by potassium dichromate, whereas, pol beta over expression can help cells response to DNA damage and protect cells from death in a certain degree.
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ADN Polimerasa beta/fisiología , Reparación del ADN/genética , Fibroblastos/metabolismo , Mutación , Animales , Proliferación Celular , Células Cultivadas , Daño del ADN , ADN Polimerasa beta/biosíntesis , Embrión de Mamíferos , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Ratones , Dicromato de Potasio/toxicidadRESUMEN
OBJECTIVE: To determine the oxidative damage of mainstream smoke (MS) on Human bronchial epithelial cells (HBE) and its role in lung cancer. METHODS: MTT assay was used to test the cytotoxicity of MS on HBE. The HBE cells were treated with different concentrations of MS for 12 h. The chromatosome damage and DNA strand breaks were measured by micronucleus test and alkaline comet assay respectively. The contents of ROS in the HBE cells were determined using fluorescence method. RESULTS: With the increase of MS, the viability of HBE cells decreased. The IC50 decreased with the increasing exposure time to MS, showing significant dose-effect and time-effect relationships. The MS induced DNA strand break in the HBE cells. The comet, L Tail, Tail DNA and OTM increased with the increase of MS concentrations. Cigarette smoke also induced chromosome damage. The micronucleus rate of the HBE cells exposed to more than one cigarette/L of MS was significantly greater than the controls (P<0.05). The ROS increased with the concentration of MS. CONCLUSION: MS induces ROS in HBE cells, resulting in increased cytotoxicity, chromosome damage and DNA strand breaks, which suggests that oxidative damage is an important mechanism of lung cancer caused by MS.