ABSTRACT
The objective of this work is to synthesize indolacin-5-fluorouracil-1-ylmethyl ester and the structure was confirmed by means of UV, IR, [1]H-NMR, [13]C-NMR and mass spectrometry. The physicochemical parameters of melting point, solubility, apparent partition coefficient were investigated. S180 sarcoma, H22 hapatitic cancer and Lewistransplanted mice were used to evaluate the anti-tumor activity of indolacini-5-fluorouracil-1-ylmethyl ester compared with 5-fluorouracil in vivo. Anti-inflammatory and analgesic activities were evaluated in mice. The inhibitory ratio of indolacini- 5-fluorouracil-1-ylmethyl ester is comparative to that of 5-fluorouracil. This study indicates that 5- fluorouracil-1-ylmethyl ester may represent a new anticancer predrug of 5-fluorouracil to produce a combined effect of indolacin and 5-fluorouracil for cancer therapy
ABSTRACT
<p><b>OBJECTIVE</b>To establish a new method for sperm sorting by imitating the physiological process of sperm-cervical mucus interaction on the microfluidic chip.</p><p><b>METHODS</b>We designed a microfluidic chip to imitate the physiological process of natural sperm sorting in the microchannel based on the interaction between sperm and cervical mucus, and obtained motile sperm after the interaction. Meanwhile, we established an integrated real-time sperm detection reservoir on this chip to determine sperm parameters using the computer-assisted sperm analysis system. We analyzed 30 samples using both microfluidic and swim-up methods, and compared the results with those obtained before sorting.</p><p><b>RESULTS</b>The rate of grade a + b sperm, the rate of morphologically normal sperm, straight-line velocity (VSL), average path velocity (VAP) and straightness (STR) were (29.78 +/- 11.24)%, (8.00 +/- 5.19)%, (18.89 +/- 4.90) microm/s, (26.84 +/- 5.13) microm/s and (70.15 +/- 7.61)%, respectively, before sorting, (71.65 +/- 11.18)%, (14.95 +/- 6.79)%, (24.14 +/- 5.95) microm/s, (32.61 +/- 6.36) microm/s and (73.87 +/- 9.34)%, respectively, after swim-up sorting, and (92.37 +/- 6.33)%, (23.33 +/- 7.67)%, (34.03 +/- 16.78) microm/s, (38.73 +/- 16.40) microm/s and (84.91 +/- 12.56)%, respectively, after sorting on the microfluidic chip. The sperm parameters obtained before sorting showed statistically significant differences from those obtained on the chip (P < 0.01) and by the swim-up method (P < 0.05).</p><p><b>CONCLUSION</b>Imitation of the physiological interaction between sperm and cervical mucus on the microfluidic chip helped the realization of both the natural sorting and real-time analysis of sperm. The quality of the sperm sorted on the microfluidic chip is significantly better than that of the sperm before sorting and sorted by the swim-up method. This has prepared the ground for imitating the fertilization process under the physiological condition on the microfluidic chip.</p>
Subject(s)
Humans , Male , Cell Movement , Cell Separation , Cervix Mucus , Microfluidic Analytical Techniques , Microfluidics , Methods , Oligonucleotide Array Sequence Analysis , Semen Analysis , Sperm Motility , Physiology , Spermatozoa , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of a microfluidic sperm sorter on the routine parameters and DNA integrity of human sperm.</p><p><b>METHODS</b>We divided 40 semen samples into two aliquots and performed sperm sorting using a self-made polydimethylsiloxane microfluidic sperm sorter and the swim-up method, respectively. Then we evaluated and compared the effects of these two methods on the sperm routine parameters and DNA integrity by computer-assisted sperm analysis and sperm chromatin dispersion test.</p><p><b>RESULTS</b>After processing, sperm motility, normal morphology and tail hypoosmotic swelling rate were significantly improved, while sperm DNA damage remarkably decreased (P < 0.01). The microfluidic sperm sorter achieved a significantly lower rate of sperm DNA damage than the swim-up method ([ 8.4 +/- 5.8 ]% vs [16.4 +/- 9.2] %, P < 0.01), but no statistically significant differences were found in all other parameters between the two methods.</p><p><b>CONCLUSION</b>High-quality sperm with less DNA integrity damage could be obtained in sperm sorting with the microfluidic sperm sorter.</p>
Subject(s)
Adult , Humans , Male , Middle Aged , Cell Separation , Methods , DNA , DNA Damage , Infertility, Male , Genetics , Microfluidics , Protein Array Analysis , Semen Analysis , Methods , Sperm Motility , SpermatozoaABSTRACT
A multiplex RT-PCR assay based on GeXP system was developed in order to detect simultaneously human enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) and other coxsackieviruses (CVA4, 5, 9 and 10, CVB1, 3 and 5). Enterovirus detection was performed with a mixture of 12 pairs of oligonucleotide primers including one pair of published primers for amplifying all known pan-enterovirus genomes and eleven primer pairs specific for detection of the VP1 genes of EV71, C A16, CVA4, CVA5, CVA9, CVA10, CVB1, CVB3 and CVB5, respectively. The specificity of multiplex RT-PCR system was examined using enterovirus cell cultures and positive strains identified previously from hand-foot-and-mouth disease (HFMD) patients. Serial dilution of titrated EV71 and C A16 cell cultures and in vitro transcripted RNA of enterovirus VP1 regions were used to detect the sensitivity of the multiplex RT-PCR system. The limit of detection for this multiplex RT-PCR system was 10(0.5) TCID50/microL for EV71 and C A16 cell cultures and 1000 copies for in vitro transcripted RNA of nine viruses per assay. This multiplex RT-PCR assay is a rapid, sensitive and specific assay for the diagnosis of common enterovirus infection in cases of HFMD outbreak and is also potentially useful for molecular epidemiological investigation.
Subject(s)
Humans , DNA Primers , Genetics , Enterovirus , Classification , Genetics , Hand, Foot and Mouth Disease , Diagnosis , Virology , Reverse Transcriptase Polymerase Chain Reaction , MethodsABSTRACT
The objective of this study was to investigate the synergistic effect of soluble human recombinant tumor necrosis factor related apoptosis inducing ligand (TRAIL) protein combined with anti-vascular endothelial growth factor (anti-VEGF) antibody on inducing apoptosis of leukemia K562 cells. The inhibitory rates and apoptotic rates of K562 cells treated with TRAIL and anti-VEGF antibody alone and their combination for 48 hours were examined by CCK-8 assay and flow cytometry respectively. The results indicated that the apoptotic rates of K562 cells induced with 75, 100 and 150 ng/ml TRAIL after culture for 48 hours were (4.26±0.67)%, (8.91±0.55)% and (11.71±0.78)% respectively. The apoptotic rates of K562 cells induced with 2.5, 5 and 7.5 µg/ml anti-VEGF antibody after culture for 48 hours were (3.95±0.69)%, (7.98±0.74)% and (10.26±0.83)% respectively. The apoptotic rates of K562 cells treated with combination use of 2.5 µg/ml anti-VEGF antibody and 75 ng/ml TRAIL, 5 µg/ml anti-VEGF antibody and 100 ng/ml TRAIL, and 7.5 µg/ml anti-VEGF antibody and 150 ng/ml TRAIL for 48 hours were (22.16±0.93)%, (36.32±1.31)% and (49.19±0.71)% respectively. The combined use of above mentioned agents induced significantly higher apoptosis and cytotoxicity than that of TRAIL or anti-VEGF antibody alone (p<0.05). It is concluded that the combination use of TRAIL and anti-VEGF antibody can significantly increase the sensitivity of K562 cells to apoptosis.
Subject(s)
Humans , Antibodies, Monoclonal , Pharmacology , Apoptosis , K562 Cells , TNF-Related Apoptosis-Inducing Ligand , Pharmacology , Vascular Endothelial Growth Factor A , Allergy and ImmunologyABSTRACT
Objective To develop an effective way to evaluate the accurate platelet count in a patient with anticoagulants-induced pseudothrombocytopenia (PTCP).Methods It was studied that various anticoagulants effect on the platelets count for an infrequent patient with anticoagulants-dependent PTCP. When vitamin B6,aminophylline,gentamicin and amikacin were separately added to four anticoagulated blood samples from anticoagulants-dependent patient within 15 min after blood withdrawal,platelets count and morphological changes of blood cells after 4 hours of incubation at room temperature were investigated. The best anti-aggregating agent and its optimal concentration among them were explored.Results The four anticoagulants all could not inhibit the aggregation of the patient's platelets.Only amikaein among the above anti-aggregating agents can prevent and dissociate the aggregation of platelets without apparent morphological changes of blood cells and the platelet counts was stable within 4 hours after blood drawn when amikacin was added either before or after blood sampling.With increasing the concentration of amikaein,the platelet counts increase and then tend to be stable.The optimal concentration of amikacin is 5 mg/ml blood.Conclusions The supplementation of amikaein either before or after blood sampling is a useful method for the diagnosis anticoagulants-dependent PTCP and for the eva/uation of platelet counts in infrequent patients with anticoagulants-dependent PTCP.