Systematical studies are lacking on the influencing factors and mechanisms of the heparin enhanced sperm capacitation, although many studies have shown that heparin enhanced sperm capacitation. The effect of heparin concentration and exposure time, incubation temperature and co-culture with oviductal epithelial cells or cumulus cells on goat sperm capacitation were investigated in this study. The motility, membrane and acrosome integrity and capacitated percentage of goat spermatozoa were assessed after different heparin treatments, and rates of fertilization and embryo cleavage were compared after in vitro insemination of oocytes with spermatozoa capacitated by different heparin treatments. The major results are summarized as follows: 1) When spermatozoa were capacitated with heparin at 5, 10, 25, 50 and 100 microg/mL for 45 min, 50 and 100 microg/mL heparin treatments produced the highest capacitated percentages of 55% and 56%, respectively, but the percentage of spermatozoa with intact acrosomes in the 100 microg/mL heparin treatment decreased significantly (P < 0.05) in comparison with that in the control group, indicating that the optimal heparin concentration for goat sperm capacitation would be 50 microg/mL. 2) Capacitated percentage of spermatozoa increased with extension of treatment time when goat sperm were treated with 50 microg/mL heparin for 0, 10, 20, 30, 45, 60 or 120 min. Although heparin treatments for 45 to 120 min did not differ significantly (P > 0.05) in capacitated sperm percentages, sperm motility and membrane integrity decreased significantly when treated with heparin for 120 min. This suggested that the optimal exposure time of heparin at 50 microg/mL for goat sperm capacitation would be 45 to 60 min. 3) Significantly higher capacitated percentages of spermatozoa were obtained when goat sperm were treated at 42 and 38.5 degrees C than at 15 and 37 degrees C, but sperm motility and acrosome integrity were significantly lower when spermatozoa were treated at 42 degrees C than they were treated at other temperatures. Temperature of 38.5 degrees C would, therefore, be the optimal temperature for goat sperm capacitation. 4) The capacitated percentage of spermatozoa was significantly higher when goat sperm were co-cultured with oviductal epithelial cells than when treated with heparin alone or co-cultured with cumulus cells, but sperm motility and membrane and acrosome integrity did not differ significantly among the three treatments. Rates of fertilization (91.3%) and cleavage (72.2%) were significantly higher in the oviductal epithelial cell co-culture group than those in the heparin alone group. This indicated that co-culture with oviductal epithelial cells significantly enhanced goat sperm capacitation by heparin treatment.
Acrosome Reaction ; drug effects ; physiology ; Animals ; Coculture Techniques ; Epithelial Cells ; cytology ; Fallopian Tubes ; cytology ; Female ; Fertilization in Vitro ; Goats ; Heparin ; pharmacology ; Male ; Sperm Capacitation ; drug effects ; physiology ; Sperm Motility ; Spermatozoa ; cytology ; physiology

OBJECTIVETo evaluate the feasibility and curative effect of thymectomy for myasthenia gravis (MG) by video-assisted thoracoscopic surgery (VATS) through right anterior-lateral approach.

METHODSFifty-six patients of MG were treated with thoracoscopic thymectomy and mediastinal fat dissection through right anterior-lateral approach from August 2001 to October 2007. The feasibility, safety, complication and remission for MG were retrospectively analyzed.

RESULTSFifty-five operations were completed by VATS. The mean operative time and blood loss were (96.2 +/- 52.1) min and (68.7 +/- 21.4) ml, respectively. The brachiocephalic vein injury by the electric coagulator occurred in two cases and one of them performed thoracotomy for homeostasis, the other performed ligation. The postoperative pathology showed hyperplasia in 38 cases, atrophy in 5 cases, thymoma in 12 cases and cyst of thymus in 1 case. And the operative complication included one myasthenia crisis (1.8%) at the third day and one death (1.8%) at the eighth day because of postoperative hemorrhage. The average length of stay was (7.9 +/- 2.9) d. All cases were followed up from one to seventy months. Eight (14.3%) of complete remission, 39 cases (69.6%) of partial remission and 7 cases (12.5%) of no change were found. The total effective rate was 83.9%.

CONCLUSIONSThoracoscopic thymectomy through right anterior lateral approach is technically feasible, safe and minimally invasive. It has a high remission rate for MG.

Adolescent ; Adult ; Aged ; Child ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Myasthenia Gravis ; surgery ; Retrospective Studies ; Thoracic Surgery, Video-Assisted ; Thymectomy ; methods ; Treatment Outcome

This study aimed to reveal the pathogen spectrum of hand-foot-and-mouth disease (HFMD) and genetic characteristics of Coxsackievirus A16 (CoxA16) isolates in Beijing in 2009. From 1044 clinical specimens collected from 975 HFMD cases at Beijing Pediatrics Hospital, Beijing You'an Hospital and Beijing Ditan Hospital in 2009, viral nucleic acids of enterovirus were detected by reverse transcriptase polymerase chain reaction (RT-PCR). Enterovirus isolations were conducted with rhabdomyosarcoma (RD) cell line on 200 throat swabs having positive RT-PCR results. Sequencing and analyses of VP1 encoding gene were performed on 9 CoxA16 isolates in this study. The results showed that CoxA16 (49.4%) and EV71 (36.4%) were the major pathogens for the epidemics of HFMD in 2009 in Beijing, and CoxA16 was the predominant serotype, while there were also other enterovirus co-circulating, such as CoxA4, CoxA10, and CoxA9; the CoxA16 strains prevalent in Beijing in 2009 belonged to subgenotype B1a and B1b.
Child ; Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny

BACKGROUNDChemokine-like factor 1 (CKLF1) was recently identified as a novel cytokine. The full-length CKLF1 cDNA contains 530 bp encoding 99 amino acid residues with a CC motif similar to that of other CC family chemokines. Recombinant CKLF1 exhibits chemotactic activity on leucocytes and stimulates proliferation of murine skeletal muscle cells. We questioned whether CKLF1 could be involved in the pathogenesis of inflammation and proliferation in the lung. Therefore we used efficient in vivo gene delivery method to investigate the biological effect of CKLF1 in the murine lung.

METHODSCKLF1-expressing plasmid, pCDI-CKLF1, was constructed and injected into the skeletal muscles followed by electroporation. Lung tissues were obtained at the end of week 1, 2, 3 and 4 respectively after injection. The pathological changes in the lungs were observed by light microscope.

RESULTSA single intramuscular injection of CKLF1 plasmid DNA into BALB/c mice caused dramatic pathological changes in the lungs of treated mice. These changes included peribronchial leukocyte infiltration, epithelial shedding, collagen deposition, proliferation of bronchial smooth muscle cells and fibrosis of the lung.

CONCLUSIONSThe sustained morphological abnormalities of the bronchial and bronchiolar wall, the acute pneumonitis and interstitial pulmonary fibrosis induced by CKLF1 were similar to phenomena observed in chronic persistent asthma, acute respiratory distress syndrome and severe acute respiratory syndrome. These data suggest that CKLF1 may play an important role in the pathogenesis of these important diseases and the study also implies that gene electro-transfer in vivo could serve as a valuable approach for evaluating the function of a novel gene in animals.

Animals ; Base Sequence ; Bronchoalveolar Lavage Fluid ; cytology ; Cell Movement ; Chemokines ; genetics ; physiology ; Electroporation ; Humans ; Lung ; pathology ; MARVEL Domain-Containing Proteins ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Plasmids ; Pulmonary Fibrosis ; etiology

Prohibitin (PHB), an evolutionarily conserved mitochondrial inner membrane protein, is highly expressed in cells that require strong mitochondrial function. Recently, we demonstrated that the deletion of Phb in spermatocytes results in impaired mitochondrial function. In addition, PHB expression in the mitochondrial sheath of human sperm has a significantly negative correlation with mitochondrial reactive oxygen species levels, but a positive one with mitochondrial membrane potential and sperm motility. These results suggest that mitochondrial PHB expression plays a role in sperm motility. However, the mechanism of PHB-mediated regulation of sperm motility remains unknown. Here, we demonstrate for the first time that PHB interacts with protein kinase B (AKT) and exists in a complex with phospho-PHB (pT258) and phospho-AKT in the mitochondrial sheath of murine sperm, as determined using colocalization and coimmunoprecipitation assays. After blocking AKT activity using wortmannin (a phosphatidylinositol 3-kinase [PI3K] inhibitor), murine sperm have significantly ( P < 0.05) decreased levels of phospho-PHB (pT258) and the total and progressive motility. Furthermore, significantly ( P < 0.05) lower levels of phospho-PI3K P85 subunit α+γ (pY199 and pY467) and phospho-AKT (pS473; pT308) are found in sperm from infertile asthenospermic and oligoasthenospermic men compared with normospermic subjects, which suggest a reduced activity of the PI3K/AKT pathway in these infertile subjects. Importantly, these sperm from infertile subjects also have a significantly ( P < 0.05) lower level of phospho-PHB (pT258). Collectively, our findings suggest that the interaction of PHB with AKT in the mitochondrial sheath is critical for sperm motility, where PHB phosphorylation (pT258) level and PI3K/AKT activity are key regulatory factors.