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Hydrogen peroxide (H2O2) and nitric oxide (NO) has a short half-life, low bioavailability, poor tumor targeting and systemic adverse reactions in the physiological environment. In this study, phacoemulsification and nano-precipitation were used to synthesize didecyl dimethyl ammonium bromide (DDAB)/polylactic acid nanoparticles (PLA), then L-arginine (L-Arg) and glucose oxidase (GOx)-loaded nanoparticles (GADP) were prepared, and the in vitro antitumor activity was investigated．The particle size, potential, embedding rate and the ability to produce H2O2/NO of the nanoparticles were investigated. Meanwhile, in vitro cell cytotoxicity against human hepatoma cells (HepG2) was evaluated．The results showed that the prepared L-Arg-DDAB/PLA (ADP) nanoparticles were spherical particles. And the particle size and zeta potential were (225.7 ± 6.33) nm and (+23.5 ± 0.12) mV, respectively. The adsorption rate of GOx was 87.23% ± 0.02%. The drug loading of L-Arg was 15.6% ± 0.22%. The pH value of glucose solution and the amount of H2O2 showed that GADP had good catalytic activity. In vitro cytotoxicity experiments showed that blank nanoparticles were nontoxic, while the drug-loaded nanoparticles presented enhanced antitumor effect on HepG2 cells. And can inhibit tumor cell migration. The low dose nano-scale NO delivery system GADP can effectively inhibit the migration of tumor cells and kill tumor cells, thus producing therapeutic benefits.
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Objective:To select and evaluate new Chinese herbal prescription for the treatment of decreased ovarian reserve （DOR） and its appropriate dosage. Method:The literature concerning the treatment of DOR with traditional Chinese medicine （TCM） was retrieved from such databases as Chinese Journal Full-text Database， Wanfang Data Knowledge Service Platform， and Chongqing Weipu Database for Chinese Technical Periodicals （VIP）， based on which a database was established using the Traditional Chinese Medicine Inheritance Support System （TCMISS） V2.5. The data mining was then carried out to obtain the core combinations of Chinese herbs and new Chinese herbal prescription combinations， followed by the determination of the new Chinese herbal prescription by expert group discussion for experiment evaluation. The female SD rats were divided into the normal group， DOR model group， Kuntai capsule group， and low-， medium-， and high-dose new Chinese herbal prescription groups， with 12 rats in each group. Rats in the Kuntai capsule group and low-， medium-， and high-dose new Chinese herbal prescription groups were treated with Kuntai capsule solution （0.5 g·kg<sup>-1</sup> determined according to the dosage in the instruction） and 3.037 5， 6.075， and 9.12 g·kg<sup>-1</sup> new Chinese herbal prescription， respectively. After 21 days， the estrous cycle was observed by vaginal exfoliated cell smear， and the ovarian structure was observed by hematoxylin-eosin （HE） staining. The serum anti-mullerian hormone （AMH）， follicle stimulating hormone （FSH）， luteinizing hormone （LH）， and estrogen （E<sub>2</sub>） contents as well as the reactive oxygen species （ROS）， malondialdehyde （MDA）， and glutathione （GSH） levels and superoxide dismutase （SOD）， catalase （CAT）， and glutathione peroxidase （GSH-Px） activities in ovary were detected using biochemical methods. Result:The new Chinese herbal prescription subjected to experimental evaluation was composed of 11 Chinese herbs， namely Rehmanniae Radix Praeparata 20 g， Cervi Cornus Colla 12 g， Lycii Fructus 20 g， Corni Fructus 12 g， Albiziae Cortex 9 g， Nelumbinis Plumula 3 g， Salviae Miltiorrhizae Radix et Rhizoma 20 g， Astragali Radix 30 g， Atractylodis Macrocephalae Rhizoma 12 g， Dioscoreae Rhizoma 30 g， and Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle 6 g. Compared with the model group， the Kuntai capsule group and medium- and high-dose new Chinese herbal prescription groups exhibited significantly improved estrous cycle and follicular development， elevated serum AMH and E<sub>2</sub> and ovarian GSH （<italic>P</italic><0.05）， decreased serum FSH and LH （<italic>P</italic><0.05） and ovarian ROS and MDA （<italic>P</italic><0.05）， and enhanced SOD， CAT， and GSH-Px activities （<italic>P</italic><0.05）. There were no significant differences in the above-mentioned indexes between the Kuntai capsule group and the middle- and high-dose new Chinese herbal prescription groups， but the estrous cycle and follicular development were better in the latter two groups. Conclusion:The new Chinese herbal prescription screened by data mining is able to enhance ovarian antioxidation， promote follicular development， ameliorate serum hormone and estrous cycle， and effectively improve ovarian reserve function in DOR rats. The medium dose （6.075 g·kg<sup>-1</sup>） has been proved optimal.
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Due to the advantages of polymer micelles and the anticancer activity of doxorubicin (DOX), the polymer micelle of DOX is expected to be used for drug delivery in anticancer applications. As a biocompatible and biodegradable polymer, amphiphilic copolymer heparosan-adipic dihydrazide-vitamin E succinate (KV) can be self-assembled to form micelles with core-shell structure in aqueous phase. In this article, KV conjugates with two different degrees of substitution (DS) were synthesized to load DOX and were characterized by 1H NMR. The size distribution, morphology, zeta potential and release behavior in vitro of the DOX-loaded micelles were studied. In vitro cytotoxicity was investigated by MTT assay against MGC80-3 and COS7 cells. The cellular uptake of the DOX-loaded micelles was observed by fluorescence microscopy and flow cytometry. The 1H NMR spectra results confirmed the KV polymers were successfully conjugated and the degree of VES grafted on heparosan polysaccharide were 12% and 25%. Briefly, the micelles with two different DS were expressed as KV12 and KV25. The DOX-loaded micelles could resist serum adsorption because of the negative charge on the surface. The average particle size measured by dynamic light scattering (DLS) method was 140-150 nm and the TEM results indicated that the morphology of DOX-loaded micelles were spherical. The encapsulation efficiency and drug loading were 80% and 10%-15%, respectively. The DOX-loaded micelles had sustained release behavior and the cumulative release of DOX/KV12 was slightly higher than DOX/KV25. Moreover, the viabilities of cells which were co-incubated with blank micelles were greater than 90%. It is clear that the blank micelles almost non-toxic to both cells. The IC50 of drug-loaded micelles against COS7 cells was much higher than that of MGC80-3 cells and the DOX/KV12 exhibited greater cytotoxicity. The cellular uptake of DOX/KV on MGC80-3 was greater than COS7 cells. In this study, KV polymer micelles have a sustained drug release activity and have a good selectivity to tumor cells, so it would be a potential carrier in drug delivery.
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BACKGROUND:The traditional corneal scaffolds exhibit poor strength and biological compatibility. Little is reported on the artificial cornea prepared by collagen and chondroitin sulfate (CS), which consist of the natural corneal tissue. OBJECTIVE:To prepare the collagen/CS/fibroblast growth factor (FGF) composite artificial cornea with slow-release growth factor, high strength and light transmittance, as well as good biocompatibility. METHODS:Regenerated collagen films were prepared by 1％, 5％, 10％ collagen solutions using flow casting method, and the regenerated collagen film with the best bioactivity that was prepared by 5％ collagen solution was screened through a biomechanical test. Then, the CS/collagen composite film was achieved by cross-linking the CS (2, 20, 80 g/L) with collagen by using N-(3-Dimethylaminopropyl)- N'S-ethylcarbodimide hydrochloride-N-Hydroxysuccinimide. The composite film made of 20 g/L CS was confirmed to have the best transparency, which was used to be mixed with 5, 25, 50 mg/L FGF in PBS for 24 hours to prepare the collagen/CS/FGF composite films. ELISA method was used to detect the FGF level in the supernatant. Afterwards, corneal epithelial cells were co-cultured with regenerated collagen film, collagen/CS composite film and collagen/CS/FGF composite film, respectively. After 48 hours of co-culture, cell proliferation was detected by MTT method, based on which we could screen the optimal collagen/CS/FGF composite film. After co-culture with the collagen/CS/FGF composite film for 48 and 72 hours, cell morphology was observed by confocal microscope and scanning electron microscope, respectively. RESULTS AND CONCLUSION:The release amount of FGF from the composite films was dependent on the initial loading amount of FGF. Meanwhile, FGF released slowly from the three kinds of composite films, and the release amount was 11％, 23％, 30％ at 72 hours after culture, in accordance with the pharmacokinetic process. MTT findings indicated that the optimal loading concentration of FGF was 25 mg/L. Under the microscope, the collagen/CS/FGF composite film promoted the adhesion, growth and proliferation of corneal epithelial cells. To conclude, the collagen/CS/FGF composite film is expected to be an ideal scaffold material for artificial cornea preparation.
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[Objective]We aimed to explore the diagnostic value of DNA fragmentation index(DFI)and acrosin activi-ty in male infertility.[Methods]Nine hundred and two semen samples were collected from patients and assessed for the DNA fragmentation index by sperm chromatin dispersion test,acrosin activity,as well as standard sperm parameters according to the WHO criteria. Statistical analysis was performed with SPSS.[Results]Statistically significant differences were observed in age,sperm concentration,total motility,progressive rate and acrosin activity among different group of sperm DNA dam-age(≤10%,11~20%,21~30%,≥31%).Sperm acrosin activity also showed difference in sperm concentration,total num-ber,total motility,progressive rate,normal morphology rate,teratozoospermia index(TZI)and sperm DFI.The DNA frag-mentation rate and sperm concentration,sperm motility,forward motility rate,total sperm count and acrosin activity was neg-atively correlated,while it is negative correlatd with TZI. Acrosin activity and sperm concentration,sperm motility,forward motility rate,sperm count and normal morphology rate was positively correlated,while it is negative correlated with the ab-normal rate of head,sperm deformity index,DNA fragmentation rate.[Conclusion]Sperm DNA damage and acrosin activity could partly reflect the quality of sperm. Moreover,sperm DFI maypredict the sperm motility part while the acrosin activity more likely related to sperm morphology.
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In this study, the endocytosis pathway of heparosan and its intracellular distribution were investigated in MCF-7 tumor cells and COS7 normal cells. The endocytosis inhibition and cellular probe location experiments showed that MCF-7 tumor cells took heparosan more efficiently and selectively than COS7 cells. The cellular uptake of heparosan was energy-dependent in both MCF-7 tumor cells and COS7 normal cells. Moreover, the major endocytosis pathway of heparosan into MCF-7 tumor cells was caveolin-mediated endocytosis and macropinocytosis. The internalized heparosan was mainly located in lysosomes of the cells.
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<p><b>OBJECTIVE</b>To develop a novel colorimetric method for detecting the tumor biomarker vascular endothelial growth factor (VEGF) based on aptamer and magnetic beads.</p><p><b>METHODS</b>The capture aptamer was hybridized to urease functionalized single-stranded DNA (ssDNA) and immobilize on the surface of magnetic beads by specific biotin-avidin binding. In the presence of VEGF, aptamers bound to VEGF to form a specific stem-loop structure to release the urease functionalized ssDNA. After separation, the supernatant was transferred to a tube and urea and phenol red were added. Urease hydrolyzed urea to produce ammonia to cause an increase of the pH value and a color change of phenol red. The results were inspected with either the naked eyes or by a UV spectrophotometer.</p><p><b>RESULTS</b>Under optimized conditions, the detection system showed a good linear relationship for VEGF detection in the range of 0.1 to 10 pmol/L with a detection limit as low as 0.06 pmol/L. The results of VEGF detection in the serum of patients with lung cancer were consistent with those using an ELISA Kit. The results of examination of 10 serum samples with this aptamer-based method and ELISA kit showed that the accuracy of this method was 90%.</p><p><b>CONCLUSION</b>This aptamer-based system provides an simple and convenient method for VEGF detection with a high sensitivity and selectivity.</p>
Subject(s)
Humans , Aptamers, Nucleotide , Biomarkers, Tumor , Colorimetry , DNA, Single-Stranded , Lung Neoplasms , Nucleic Acid Hybridization , Vascular Endothelial Growth Factor AABSTRACT
<p><b>OBJECTIVE</b>To evaluate male serum anti-Müllerian hormone (AMH) in assessing semen quality and predicting the pregnancy outcomes of assisted reproductive technology (ART).</p><p><b>METHODS</b>A total of 103 male patients under ICSI were allotted to Groups A (normal sperm concentration control, n = 29), B (oligospermia, n = 27), C (obstructive azoospermia, n = 29) and D (non-obstructive azoospermia, n = 18). The contents of serum AMH and other related sexual hormones were determined by ELISA, and their correlations were analyzed with the seminal quality on the day of semen collection and with the pregnancy outcomes after ICSI.</p><p><b>RESULTS</b>The contents of male serum AMH were (5.03 +/- 0.44), (3.70 +/- 0.44), (5.39 +/- 0.71) and (7.31 +/- 1.64) pmol/L, respectively, in Groups A, B, C and D, with no statistically significant differences among the four groups (F = 2.02, P > 0.05). The egg fertilization rate of the 103 couples was (76.13 +/- 23.66) %, not significantly correlated with the male serum AMH level (P > 0.05). The contents of male serum AMH in the pregnancy and non-pregnancy groups were (6.19 +/- 1.05) and (4.72 +/- 1.64) pmol/L, respectively, with no significant difference (t = 1.281, P > 0.05).</p><p><b>CONCLUSION</b>The level of male serum AMH can neither reflect spermatogenesis of men nor predict the egg fertilization rate and pregnancy outcomes after ICSI, and therefore cannot be used alone as a serological predictive marker of ICSI outcomes.</p>
Subject(s)
Female , Humans , Male , Pregnancy , Anti-Mullerian Hormone , Blood , Case-Control Studies , Pregnancy Outcome , Prognosis , Semen Analysis , Sperm Injections, IntracytoplasmicABSTRACT
<p><b>OBJECTIVE</b>To determine a dengue fever outbreak in Yiwu city, Zhejiang Province in 2009 and to trace the origin of the pathogen.</p><p><b>METHODS</b>The dengue virus IgM, IgG antibodies and viral nucleic acid were detected and virus was isolated using 40 serum samples from the suspected patients. The viral RNA of the isolated virus strains was extracted and the E gene was amplified by RT-PCR. The amplicons were sequenced and the phylogenetic and homological analyses were also constructed.</p><p><b>RESULTS</b>Among 40 serum samples from dengue fever suspected patients, 17 were positive from for dengue IgM (42.5%); 4 were IgG positive (10.0%); 34 samples were dengue virus RNA positive (85.0%), 28 dengue virus type 3 (D3) strains were isolated (70.0%). The complete coding region of envelope genes (E) from 13 D3 strains was all 1479 nt without any insertion or deletion, which encoded with 493 amino acids (aa). E gene from the 13 D3 strains from Zhejiang in 2009 (D3/ZJ/2009) was 100.0% identical. The strain from Saudi Arabia shared the highest similarity with the D3 strain, 99.3% and 100.0% of their E genes and deduced amino acids were identical, respectively, whereas they were 93.4% and 97.4% between D3/ZJ/2009 strain and its prototype strain (D3/H87/1956), and 93.6% and 97.4% between D3/ZJ/2009 and a D3 strain isolated in Guangxi Province in 1980. The phylogenetic tree of E genes also indicated that D3/ZJ/2009 had maximum similarity with the D3/Saudi Arabia/2004. They all belonged to the D3/GIII branch, which was originated from Indian Subcontinent.</p><p><b>CONCLUSION</b>The outbreak of dengue fever in Zhejiang in 2009 was caused by type 3 dengue virus III genotype. The virus was most likely originated from Saudi Arabia.</p>
Subject(s)
Adolescent , Adult , Aged, 80 and over , Female , Humans , Male , Middle Aged , China , Epidemiology , Dengue , Epidemiology , Virology , Dengue Virus , Classification , Genetics , Disease OutbreaksABSTRACT
Objective To investigate the impacts of body mass index (BMI) and age on in vitro fertilization-embryo transfer (IVF) and intracytoplasmic sperm injection (ICSI) treatment in infertile patients without polycystic ovary syndrome (PCOS). Methods A retrospective study of 1426 patients during Jun. 2001 - Nov. 2009 was carried out. Multiple regression was used to analyze the effects of BMI (low weight: BMI＜18.5 kg/m2, normal weight:BMI 18.5-23.99 kg/m2 and over weight-obesity: BMI≥24 kg/m2) and age (young: 20-34 years old, eld: 35-45 years old) on controlled ovarian stimulation (COH)[including:dose and duration of Gn, E2 level on day of human chorionic gonadotropin (HCG) administration, number of oocytes collected and full-grown follicles ],number of fertilization, cleavage, two-pronucleus, normal embryos and cryopreserved embryos and clinical pregnancy outcome. Results ( 1 ) Gn dose for the patients whose age were 35 and the above,had a positive correlation with age (P＜0.001), 12.70% of the total variation of Gn dose was related to age (standardized partial regression coefficient was 0.343). (2) Estradiol level on day of HCG administration had a negative correlation with BMI in overweight-obesity patients, and so were the patients whose age were 35 and above (P value respectively lower than 0.037 and 0.018). 0.80% of the total variation of estradiol (HCG day) is related to age and overweight-obesity while age took greater proportion (standardized partial regression coefficients were 0.066 and 0.058 respectively). (3)For older patients, age appeared to have negative relationships with duration of Gn and number of oocytes collected, full-grown follicles, fertilization, cleavage, two-pronucleus, normal embryos and cryopreserved embryos (P＜0.05). (4)Compared to young-normal weight patients, the odds ratio of pregnancy in eld-low weight and eld-overweight-obesity patients were 0.482 and 0.529 (P＜0.05)respectively. Conclusion Age, but not the BMI, had significant effects on IVF/ICSI treatment. It seems that factors as losing weight before IVF or ICSI treatment effective in reducing the dose of Gn.
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0.05).The scales of subtests including arithmetic,digit span,picture completion,block design and coding of children in CAE group were significantly lower than those of control group(Pa