OBJECTIVETo establish the two-dimensional electrophoresis profiles with high resolution and reproducibility from the brain tissues of mice with human cytomegalovirus (HCMV) infection.

METHODSForty Kunming mice were randomized into HCMV infection group (n=20) with HCMVAD(169) injection and control group (n=20) with saline injection in the brain. Thirty days after the injections, the brain tissue of the mice were taken and the protein fractions were isolated by two-dimensional gel electrophoresis (2-DE). Image Master 2D software was used to identify the differentially expressed proteins, and the peptide mass fingerprint (PMF) data were obtained for identification of the differential protein spots via database searching. Western blotting was performed to verify the expressions of some of the differential proteins.

RESULTS AND CONCLUSIONThe 2-D maps of the brain tissues with high Well resolution and reproducibility were obtained. Some of the differentially expressed proteins identified by mass spectrometry (MS) matched their counterparts in the SWISS-2DPAGE database. Western blotting analyses verified the differential expression of the individual proteins. These data can be of value for studying the diagnosis, pathogenesis and effective therapeutic targets of the disease.

Animals ; Animals, Newborn ; Brain ; metabolism ; virology ; Cytomegalovirus Infections ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Mice ; Proteins ; analysis ; Proteome ; analysis ; Proteomics ; methods ; Software

AIMTo evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI).

METHODSFifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration.

RESULTSA total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term.

CONCLUSIONIt appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.

Adult ; Cell Culture Techniques ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Female ; therapy ; Infertility, Male ; therapy ; Male ; Oocytes ; growth & development ; Pregnancy ; Pregnancy Rate ; Semen ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; Testis ; cytology

Brain Mapping ; Cognitive Dysfunction ; Humans ; Magnetic Resonance Imaging ; Schizophrenia/therapy* ; Transcranial Magnetic Stimulation