In vitro developmental competence of pig nuclear transferred embryos: effects of GFP transfection, refrigeration, cell cycle synchronization and shapes of donor cells.

The present study was designed to evaluate the feasibility of producing pig transgenic blastocysts expressing enhanced green fluorescent protein (GFP) and to examine the effects of shape and preparation methods of donor cells on in vitro developmental ability of pig nuclear transferred embryos (NTEs). In experiment 1, the effect of GFP transfection on development of pig NTEs was evaluated. The cleavage and blastocyst rates showed no significant difference between NTEs derived from transfected and non-transfected donors. In experiment 2, the effect of different nuclear donor preparation methods on in vitro development of NTEs was examined. The cleavage rate showed no statistically significant differences among three preparation methods. The blastocyst rates of donor cells treated once at -4 degrees C and those of freshly digested cells were similar to each other (26.3% vs 17.9%). The lowest blastocyst rates (5.88%) were observed when cells cryopreserved at -196 degrees C were used as donors. In experiment 3, the effect of different cell cycle synchronization methods on the in vitro development potential of pig NTEs was evaluated. The cleavage rate of NTEs derived from cycling cells was much better than that of NTEs derived from serum-starved cells (64.4% vs 50.5%, p < 0.05), but no significant difference was observed between the the blastocyst rates of the two groups. In experiment 4, the effect of different shapes of cultured fibroblast cells on the in vitro development of pig NTEs was examined. The fusion rate for couplets derived from rough cells was poorer than that observed in couplets derived from round smooth cells (47.8% vs 76.8%, p < 0.05). However, there were no significant differences observed in the cleavage rate and blastocyst rate. In conclusion, the present study indicated that (i) refrigerated pig GFP-transfected cells could be used as donors in nuclear transfer and these NTEs could be effectively developed to blastocyst stage; (ii) serum starvation of GFP-transfected cells is not required for preimplantation development of pig NTEs; and (iii) a rough surface of GFP-transfected donor cells affects fusion rate negatively but has no influence on the cleavage rate or blastocyst rate of pig NTEs.

[Surgery of nasopharyngeal angiofibroma].

OBJECTIVE: To study the operations of nasopharyngeal angiofibroma. METHOD: Twenty-two patients with nasopharyngeal angiofibroma operated by various operative approaches were studied in this research. All their clinic data were collected and analyzed. RESULT: In the operation, the brooding amount was from 400 ml to 1600 ml. One case was recurred in half a year after operation. The other 21 cases didn't recur after one to two years follow-up. CONCLUSION: In order to improve the treatment, reduce the recurrence of tumor and decrease brooding in operation, better preparation before surgery, reasonable surgical route,external carotid artery delegation, embolization under digital subtraction angiography(DSA) and resecting the all tumor as possible in the operation are very important

[Protective effect of estrogens replacement on apoptosis cells of nasal mocosas in ovariectomized rats].

OBJECTIVE: To study the protective effect of nylestriol on apoptosis cells of atrophic nasal mucosas in ovariectomized rats and the difference between nasal drip and gastrogavage. METHOD: Sixty rats with atrophic rhinitis were divided into four groups at random (each with 15 rats): contrary group (Cg), ovariectomized group (Og), ovariectomized + nylestriol nasal drip group (ONNDg), and ovariectomized + nylestriol by gastrogavage (ONGg). Earlier apoptosis cells of nasal mucosas taken from nasal septum were measured with flow cytometry. RESULT: After being ovariectomized, the number of apoptosis cells of mucosas increased. In ONGg, the number of apoptosis cells of mucosas increased at 15 d after operation (P < 0.05), but it recovered at 30 d and 60 d after operation. In ONGg, the number of apoptosis cells of mucosas increased at 15 d and 30 d after operation (P < 0.05), but it recovered at 60 d after operation. CONCLUSION: Estrogen replacement by gastrogavage and via nasal drip have effects on protecting cells of mucosas from lacking of estrogen by decreasing apoptosis cells in ovariectomized rats. It might cost more time to get therapeutic effectiveness via nasal drip than by gastrogavage.

[Distribution of calcitonin gene-related peptide in guinea pig cochlea].

OBJECTIVE: To observe the distribution of CGRP in guinea pig cochlea. METHOD: CGRP was labeled by immunostaning technique. Distribution of CGRP immunoreactivity was observed with light microscopy. RESULT: CGRP immunoreactivity was located in SGN, nerve fibers at osseous spiral lamina, stria vascularis, IHC, adjacent area of OHC and Deiters' cells. CONCLUSION: CGRP distributes extensively in guinea pig cochlea. CGRP may play an important role as neurotransmitter and/or neuromodulator to hearing function by modulating HC, supporting cells, SGN and other neurons along the process of auditory nerve. It may have effect on on cochlear blood flow, too.

[Protective effect of daizein on apoptosis cells of nasal mucosas in ovariectomized rats].

OBJECTIVE: To study the protective effect of daizein on apoptosis cells of atrophic nasal mucosas in ovariectomized rats. METHODS: Sixty rats were divided into four groups as contrary, ovariectomized, ovariectomized + nylestriol (O + N) and ovariectomized + daizein (O + D), each with 15 rats. Earlier apotosis cells of nasal mucosas taken from nasal septum were measured with flow cytometry. RESULTS: Compared with contrary group, the number of apoptosis cells of mucosas increased after being ovariectomized, the number of apoptosis cells of mucosas in O + N and O + D groups didn't change. CONCLUSION: Estrogen replacement and daizein might have effects on protecting cells of mucosas from lacking of estrogen by decreasing apoptosis cells in ovariectomized rats.