• Journal of Trauma and Injury
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• v.33 no.2
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• pp.74-80
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• 2020

Purpose: This study was conducted to investigate whether rapid and efficient administration of blood products was achieved and whether clinical outcomes were improved by applying a massive transfusion protocol (MTP). Methods: From January 2016 to September 2019, the medical records of trauma patients who received at least 10 units of packed red blood cells (PRBC) at Pusan National University Hospital (level I trauma center) were retrospectively reviewed. The patients treated from January 2016 to January 2018 were designated as the non-MTP group, and those treated from February 2018 to September 2019 were designated as the MTP group. Results: During the study period, 370 patients received massive transfusions. The non-MTP and MTP groups comprised 84 and 55 patients, respectively. No significant between-group differences were found in the units of PRBC (23.2 vs. 25.3, respectively; p=0.46), fresh frozen plasma (FFP) (21.1 vs. 24.4, respectively; p=0.40), and platelets (PLT) (15.4 vs. 17.0, respectively; p=0.54) administered in the first 24 hours. No statistically significant differences between the non-MTP and MTP groups were found in the FFP-to-PRBC ratio (0.9 vs. 0.94, respectively; p=0.44) and or the PLT-to-PRBC ratio (0.72 vs. 0.72, respectively; p=0.21). However, the total number of cryoprecipitate units was significantly higher in the MTP group than in the non-MTP group (7.4 vs. 15.3 units, respectively; p=0.003) and the ratio of cryoprecipitate to PRBC in the MTP group was significantly higher than in the non-MTP group (0.31 vs. 0.62, respectively; p=0.021). The time to transfusion was significantly reduced after MTP implementation (41.0 vs. 14.9 minutes, respectively; p=0.003). Conclusions: Although no significant differences were found in the clinical outcomes of patients who had undergone severe trauma, rapid and balanced transfusion was achieved after implementing the MTP.

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.127-132
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• 1999

This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, removed seminal plasma(RSP) semen and fractional semen of small dogs, and the effect of temperature and preservatio time and cryopreservation on motility of whole and removed seminal plasma semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. 1. Average sperm concentration, sperm motility and abnormal sperm rates of whole semen and RSP semen were 5.07$\pm$2.32$\times$10$^{6}$ cells/$m\ell$, 95.42$\pm$2.65%, 4.42$\pm$0.157% and 4.69$\pm$3.27~4.25$\pm$3.65$\times$10$^{6}$ cells/$m\ell$, 91.17$\pm$3.85~88.52$\pm$3.85%, 6.57$\pm$0.43~5.54$\pm$0.52%, respectively. 2. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 1st fractional semen were 0.92$\pm$0.7$m\ell$, 4.57$\pm$0.78$\times$10$^{6}$ cells/$m\ell$, 10.72$\pm$3.21% and 5.50$\pm$0.70%. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 2nd fractional semen were 2. 14$\pm$0.19$m\ell$, 2.01$\pm$0.12$\times$10$^{6}$ cells/$m\ell$, 95.44$\pm$4.21% and 4.31$\pm$0.53%. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 3rd fractional semen were 2.66$\pm$0.23$m\ell$, 2.35$\pm$0.21$\times$10$^{6}$ cells/$m\ell$, 90.71$\pm$2.63%, 6.33$\pm$0.91%, respectively. 3. Motility of whole semen and RSP semen were higher at 2$0^{\circ}C$ than at 4$^{\circ}C$ or 37$^{\circ}C$. When preservation temperature was 2$0^{\circ}C$, sperm motility were 98.32% at 1 hr, 92.15% at 5 hrs, 90.23% at 10 hrs 82.08% at 15 hrs 70.07% at 20 hrs 60.02% at 20 hrs 37.19% at 40 hrs respectively. 4. Average sperm motility of frozen 2nd fraction semen and RSP semen were 33.3$\pm$8.7, 54.7$\pm$9.5%, respectively. Sperm motility was significantly higher in frozen 2nd fraction semen and RSP semen compared with control group.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.439-449
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• 2000

This study was to investigate the expression of transgene after co-injection of spermatozoon and EGFP gene into mature oocytes in cattle. From frozen semen, spermatozoa were treated by DTT with 0.03% Tween-20, freezing and thawing or 0.02% Triton X-100 to disrupt their plasma membranes. The sperm injected oocytes were co-cultured with bovine oviduct epithelial cells in CRlaa, and expression of EGFP in embryos were observbed under epifluorescent microscope. Two pronuclei were formed in oocytes injected with sperm treated by DTT (44.6%), DTT-Tween-20 (48.4%), DTT-freezing (44.4%) and DTT-Triton X-100 (42.9%). Cleavage and blastocyst formation rates of bovine oocytes which injected with sperm treated by DTT, DTT-Tween-20, DTT-freezing, and DTT-Triton X-100 were 49.1, 58.5, 43.9, and 48.4% and 10.2, 13.0, 6.8, and 6.5%, respectively. Although the most of embryos were showing mosaicism, embryos expressing EGFP gene were observed in all treated groups. Therefore, these results indicate that membrane disrupted sperm could interact with exogenous DNA, and that this procedure may be useful to introduce foreign gene into bovine oocytes.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.2
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• pp.79-92
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• 1992

This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

• Journal of Chest Surgery
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• v.37 no.10
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• pp.817-826
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• 2004

Background: Several studies have demonstrated that conventional hypothermic cardiopulmonary bypass (CPB) causes cellular injury, abnormal responses in peripheral vascular beds and increased postoperative bleeding, whereas normothermic CPB provides protection of the hypothermic-induced effects and better cardiac recovery. The present study was prospectively performed to compare the effects of normothermic CPB to those of hypothermic CPB on the inflammatory and hematologic responses during cardiac surgery. Material and Method: Thirty-four adult patients scheduled for elective cardiac surgery were randomly assigned to hypothermic CPB (nasopharyngeal temperature $26～28^{\circ}C,$ n=17) or normothermic CPB (nasopharyngeal $temperature>35.5^{\circ}C,$ n=17) group. In both groups, cold $(4^{\circ}C)$ crystalloid cardioplegia was applied for myocardial protection. Blood samples were drawn from radial artery before (Pre-CPB), 10 minutes after starting (CPB-10) and immediately after ending (CPB-OFF) CPB. Total leukocyte and platelet counts, interleukin-6 (IL-6) level(expressed as percent to the baseline of Pre-CPB), D-dimer level, protein C and protein S activity were measured with the blood samples. The amount of bleeding for postoperative 24 hours and blood transfusion after operation were also assessed. All parameters were compared between the two groups. Result: The total leukocyte counts $(10,032\pm65/mm^3)$ and the increased ratio of IL-6 $(353\pm7.0%)$ at CPB-OFF in the normothermic group were higher than that $(7,254\pm48/mm^3$ and $298\pm7.3%)$ of the hypothermic group(p=0.02 and p=0.03). In the normothermic group, protein C activity $(32\pm3.8%)$ and protein S activity $(35\pm4.1%)$ at CPB-OFF were significantly lower than that $(45\pm4.3%$ and $51\pm3.8%)$ of the hypothermic group (p=0.04 and p=0.009). However, there were no differences in platelet counts and D-dimer concentration. In the normothermic group, the amount of bleeding for postoperative 24 hours $(850\pm23.2$ mL) and requirements for blood transfusion after operation such as packed cell $(1,402\pm20.5$ mL), fresh frozen plasma $(970\pm20.8$ mL) and platelet $(252\pm6.4$ mL) were higher than that $(530\pm21.5$ mL, $696\pm15.7$ mL, $603\pm18.2$ mL and $50\pm0.0$ mL) of the hypothermic group. Conclusion: These results indicate that normothermic CPB with cold crystalloid cardioplegia was associated with higher increase in inflammatory response, hemostatic abnormalities and postoperative bleeding problem than moderate hypothermic CPB.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.153-161
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• 2003

This study has evaluated effect of the spermatozoa incubation on the glycosidase activity and fertilizing ability in vitro in the pig. To identify sperm glycosidases specific for sugar residues found in the zona pellucida of pig oocytes, the spermatozoa were treated experimentally and assayed for activities of $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase and N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase). The glycosidases activity were higher in spermatozoa incubated for 2h than without incubation. The $\beta$-GlcNAc'ase activity was at least two-fold higher than other glycosidase regardless of spermatozoa incubation. In the same glycosidases, the activity had a tendency to increase as time of spermatozoa incubation was prolonged, but there were no differences in spermatozoa incubated during the various periods (4~24h). The percentages of spermatozoa that reached acrosome reaction were affected by glycosidases in the medium (P<0.05, for mannosidase), and were higher in spermatozoa with that than without incubation. On the other hand, the spermatozoa motility were decreased with incubation periods, but no effects by different glycosidases on the change of sperm motility during the various periods of incubation. In other experiment, the binding and penetration of pig spermatozoa were tested with oocytes matured in vitro in the presence of various glycosidase. The penetration rates were decreased with incubation of spermatozoa when oocytes were inseminated in medium with different glycosidases. These rates were higher in spermatozoa non-incubated than with incubation for 2h (P<0.05 for GlcNAc'ase; P<0.01 for control group). The sperm-zona binding rate in control group were higherthan in medium with glycosidases. In addition, the highest binding rate were obtained in medium with GlcNAc'ase. In all glycosidases, the sperm-zona binding rate in spermatozoa without incubation were higher than incubation for 2h. The significant differences were obtained in spermatozoa treated with $\alpha$-D-mannosidase (P<0.05). These results suggest that $\beta$-GlcNAc'ase is present mainly in the plasma membrane of pig spermatozoa. It was also shown that the glycosidase activity were increased in all glycosidases in spite of low sperm-zona binding rate and penetration rates by spermatozoa incubation.