• 대한수의학회지
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• 제44권3호
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• pp.469-473
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• 2004

This study was performed to investigate the effects of calcium administration on the blood coagulation mechanism through APTT in the calf. Five male calves (70~90 kg) were used in this experiment. In the control group, heparinized normal saline (1 IU/kg/min) had been infusing into the jugular vein for 100 minutes. For the analysis of calcium effects on the APTT, the same solutions had been infusing during the first 40 minutes, subsequently the solution including calcium gluconate (3.3 mg/ml/min) had been infusing for 60 minutes. Blood samples were serially collected every 10 minutes for the APTT and platelets count and every 20 minutes for the calicum level during the infusion. In the calcium-treated group, after 70 minutes the APTT ratio (APTT heparin/APTT baseline) was higher than the therapeutic range. APTT was significantly increased at 50, 60 and 70 minutes in the calcium-treated group as compared to the control group (p<0.01). In the control group, calcium level was decreased significantly after heparin infusion (p<0.01). The platelet count was gradually decreased without significant variation in the both control and calcium-treated groups. These results suggested that APTT is slightly increased in combined heparin and calcium administration.

• Archives of Plastic Surgery
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• 제47권1호
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• pp.54-61
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• 2020

Background Conventional methods of external bleeding for congested fingertip replants exhibit notable problems, including uncontrollable bleeding and unpredictable survival of the replant. We have added a local injection of heparin calcium to the routine use of systemic heparinization for inducing external bleeding. We retrospectively examined patients who underwent external bleeding using our method. Methods Local subcutaneous injections of heparin calcium were made in 15 congested replants in addition to systemic heparinization. Each injection ranged from 500 to 5,000 U. The average duration of the injections was 4.1 days. Surgical outcomes were analyzed and compared with a control group of patients who underwent external bleeding without heparin calcium. Results The overall survival rate was 93.3%, which was higher than that of the control group (83.3%), but the difference was not statistically significant (P=0.569). The survival rate for subzones I and II by the Ishikawa subzone classification was 100%, whereas it was 87.5% in subzones III and IV. No statistically significant difference was observed. The rate of partial necrosis was 0% in subzones I and II, whereas it was significantly higher (66.7%) in subzones III and IV (P=0.015). The mean total blood loss via external bleeding was 588 g in 10 fingers. No patients required blood transfusion. Conclusions Congestion of a replanted fingertip can be successfully managed without blood transfusion by our method. Although complete relief from congestion in replants in subzones I and II is achievable, there is a higher risk of partial necrosis in subzones III and IV.

• Journal of Industrial and Engineering Chemistry
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• 제68권
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• pp.220-228
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• 2018

In this study, we found a simple surface biofunctionalization of biphasic calcium phosphate (BCP) based on the high affinity between alendronate and the calcium ions of BCP, and the strong interaction between heparin and bone morphogenic protein-2 (BMP-2). The biofunctionalized BCP did not be precipitated well and display a remarkable enhancement of osteogenic activity of human adipose-derived stem cells by showing increased alkaline phosphatase (ALP), calcium deposition and osteogenic-related genes (i.e., Runx-2, ALP, osteocalcin, and osteopontin), and bone regeneration in the calvarial defect model. Therefore, this simple surface technique can be used to easily functionalize various calcium phosphates.

• The Journal of Advanced Prosthodontics
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• 제3권3호
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• pp.152-160
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• 2011

PURPOSE. This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity. MATERIALS AND METHODS. The anodized titanium discs, not coated with any material, were used as a control group. In heparinized-Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells ($^*$P<.05 and $^{**}$P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti ($^{**}$ P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-${\alpha}$ and IL-6 of proinflammatory cytokine ($^*$P<.05 and $^{**}$P<.001). CONCLUSION. The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

• 한국식품영양학회지
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• 제22권1호
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• pp.20-28
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• 2009

According to traditional oriental medicine, only non-coagulated native deer blood is said to be effective, and coagulated deer blood is ineffective. Thus, a drying and tablet-producing method for deer blood was developed to maintain its physiological and therapeutic activity, and so that after drying, it can be redissolved and protected from coagulation. Proteases such as trypsin, pepsin, chymotrypsin, and aminopeptidase were added to the deer blood indicating that it coagulated in an hour, as shown by the reference. Wax gourd extract, which is high in protease, was added to the blood resulting in anticoagulation for 31 hours. Also, additions of 1% EDTA, 0.38% sodium citrate, 0.16% calcium oxalate, 1.2% ethanol, and 0.006% heparin to the deer blood resulted in anticoagulation for 1 hour, 4 hours, 2 hours, 1 hour, and 31 hours, respectively. In an experiment using 0.19% sodium citrate plus 1% wax gourd extract, and 0.006% heparin plus 1% wax gourd extract, anticoagulation was maintained for up to 72 hours. However, since heparin can not be used in food, the deer blood tablet was made with the addition of 0.19% sodium citrate and 1% wax gourd extract, followed by freeze drying. The dissolution rate for the tablet manufactured in this manner was 96.7%. And the dissolution rates for spray-dried deer blood, vacuum-dried deer blood, and marketed deer blood tablets were 85%, 81%, and 25.5%, respectively. The composition of the tablet produced from the freeze-dried deer blood was 56.4% protein, 18.7% lactose, 1.2% amino acids, 1.0% glucose, 0.7% lipids, 180 mg/100 g of iron, 13 mg/100 g of potassium, 39.1 mg/100 g of calcium, 480 mg/100 g of sodium, 368 mg/100 g of chloride, each.

• 약학회지
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• 제41권2호
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• pp.212-219
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• 1997

Multiple forms of extracellular phospholipase $A_2$ have been detected in human amniotic fluid (HAF). When HAF was subjected to heparin-Sepharose column chromatography, phospholipase $A_2$ activity was detected in both heparin-non binding and binding fraction. The activity of heparin-non binding fraction was further purified by sequential uses of column chromatographies on butyl-Toy-opearl 650M and DEAE-Sephacel. DEAE-Sephacel fraction contained three different phospholipase $A_2$ activities (Peak I, II, III). The molecular weight of DEAE-Sephacel fraction phospholipase $A_2$ determined by SDS-PAGE were about 52KDa (Peak I). Peak II, III required micromolar $Ca^{2+}$ ion for its maximum activity, but Peak I enzyme showed calcium independent phospholipase $A_2$ activity and showed broad range of pH (6.0~10.0) optimum. All these enzymes were not recognized by a monoclonal antibody raised against phospholipase $A_2$ from human synovial fluid. These results suggest that HAF might contain multiple forms of extracellular phospholipase $A_2$, which may neither belong to the 14KDa group II phospholipase $A_2$ family nor cytosolic phospholipase $A_2$.

• The Journal of Advanced Prosthodontics
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• 제3권3호
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• pp.145-151
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• 2011

PURPOSE. This study was performed to investigate the ability of recombinant human-bone morphogenic protein-2 immobilized on a heparin-grafted bone substrate to enhance the osteoblastic functions. MATERIALS AND METHODS. The Bio-$Oss^{(R)}$, not coated with any material, was used as a control group. In rhBMP-2-Bio-$Oss^{(R)}$ group, rhBMP-2 was coated with Bio-$Oss^{(R)}$ using only deep and dry methods (50 ng/mL, 24 h). In heparinized rhBMP-2-Bio-$Oss^{(R)}$ group, dopamine was anchored to the surface of Bio-$Oss^{(R)}$, and coated with heparin. rhBMP-2 was immobilized onto the heparinized- Bio-$Oss^{(R)}$ surface. The release kinetics of the rhBMP-2-Bio-$Oss^{(R)}$ and heparinized rhBMP-2-Bio-$Oss^{(R)}$ were analyzed using an enzyme-linked immunosorbent assay. The biological activities of the MG63 cells on the three groups were investigated via cytotoxicity assay, cell proliferation assay, alkaline phosphatase (ALP) measurement, and calcium deposition determination. Statistical comparisons were carried out by one-way ANOVA test. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The heparinized rhBMP-2-Bio-$Oss^{(R)}$ showed more sustained release compared to the rhBMP-2-Bio-$Oss^{(R)}$ over an extended time. In the measurement of the ALP activity, the heparinized group showed a significantly higher ALP activity when compared with the non-heparinized groups (P<.05). The MG63 cells cultivated in the group with rhBMP-2 showed increased calcium deposition, and the MG63 cells from the heparinized group increased more than those that were cultivated in the non-heparinized groups. CONCLUSION. Heparin increased the rhBMP-2 release amount and made sustained release possible, and heparinized Bio-$Oss^{(R)}$ with rhBMP-2 successfully improved the osteoblastic functions.

• 한국발생생물학회지:발생과생식
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• 제12권3호
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• pp.267-274
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• 2008

포배의 분화는 배아의 착상에 있어 핵심적인 단계로 배아 자체 또는 생식수관에서 유래하는 조절요인의 조절을 받는다. 이들 조절요인과 포배와의 순차적인 신호의 주고 받음은 분화의 중요한 단계로 인식되고 있다. 한편, 포배기 때 자유 칼슘을 통한 신호전달경로가 포배의 분화에 중요한 축의 하나로 제안되어 왔다. Concanavalin A(Con A)가 포배의 자유 칼슘 농도 증감을 유도한다는 것을 밝혀졌으나, 포배 내 자유 칼슘 농도를 변형시켜 부화와 그 이후의 발생을 촉진하는 것으로 알려진 heparin-binding epidermal growth factor-like growth factor(HB-EGF)와는 달리 팽창 이후의 부화를 억제하였다. 따라서 본 연구에서는 착상과정에서 중요한 역할을 하는 것으로 알려진 prostaglandin $E_2(PGE_2)$가 포배의 분화에 관여하는지를 Con A와 연계하여 알아보았다. Con A는 그 처리 시간에 관계없이 1시간 처리군 그리고 계속처리군에서 팽창은 촉진하고 부화는 유의하게 억제하였다. 특히 계속처리군에서 부화율이 1시간 처리군에 비하여 유의하게 감소하였다. 또한, $PGE_2$도 포배 내 자율 칼슘 농도를 증가시켰으나 팽창과 부화를 촉진하지 않았다. 또한, $10{\mu}m\;PGE_2$ 농도에서는 부화가 억제되는 경향을 보였다. 그러나 흥미롭게도 $PGE_2$는 Con A가 처리된 포배의 부화를 촉진하였다. Con A를 전처리한 포배에 $PGE_2$를 처리할 경우 포배 내 자유 칼슘의 농도 증감이 진행됨을 공촛점현미경을 이용하여 분석할 수 있었다. 이러한 결과는 신호물질에 의해 유도된 자유 칼슘 농도의 증감이 신호물질에 따른 각기 다른 칼슘 매개로 활성화되는 신호경로를 조절하는 것을 추정할 수 있다. 또한, 순차적 신호물질 조절에 의한 자유 칼슘의 농도 증감이 포배의 분화에 있어 중요함을 제안한다.

• 한국임상수의학회지
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• 제9권2호
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• pp.373-390
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• 1992

To assay the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 13 boars(Duroc. Landrace and Yorkshire) which had been proved to be fertile in the past. then, were preserved in BWW medium or in raw state at 18$^{\circ}C$ or at room temperature. The preserved semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding). ova penetrated by sperm(penetration) and formation of a male pronucleus(pronucleus formation) and also numbers of both bound and penetrated sperm per ovum. Between BWW and TBM medium for boar sperm. no difference in the results of hamster test was obtained. The boar spermatozoa in BWW medium, BWW with caffeine, BWW with heparin, and BWW with both caffeine and heparin showed no difference in the results of hamster test. The boar spermatozoa in BWW medium containing both calcium and RSA showed considerably higher rates of sperm binding, penetration and pronucleus formation as well as higher numbers of both bound and penetrated sperm than those not containing calcium with or without BSA( p<0.01) and also the same results higher than that containing calcium without BSA( p< 0.05). The boar spermatozoa irradiated by X-ray(70 KVP, 20mA) for 3 seconds. then, maintained at 18$^{\circ}C$ for 18 hours showed considerably lower rate of sperm binding than all the other groups including the control and X-ray groups irradiated by smaller dose or maintained for shorter period(p<0.01), and also showed lower number of bound sperm than the other groups(p<0.01, p<0.05). All the control groups of both raw and diluted sperm in BWM medium showed higher rates of sperm binding, penetration and pronucleus formation as well as higher number of penetrated sperm than all the X-ray groups irradiated for 3 seconds(70KVP, 20mA) and maintained for either 3 or 18 hours (p<0.01, p<0.05). At the same time the control groups of diluted sperm showed considerably higher rates of sperm penetration and pronucleus formation than the control group of raw sperm( p<0.01). These results indicates that fertile boar sperm showed considerably lower rates In the results of hamster test, when incubated in the medium without calcium and irradiated by X-ray than when incubated in the medium with calcium and not irradiated by X-ray, respectively, to prove consequently that hamster test would be of great value in assaying the fertilizing capacity of boar spermatozoa.

• Toxicological Research
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• 제20권3호
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• pp.241-250
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• 2004

We previously found that bisphenol A (BPA) caused neurotoxic behavioral alteration. Since disturbance of calcium homeostasis is an implicated contributor in the neurotoxic mechanism of environmental toxicants, we investigated whether BPA alters calcium homeostasis. Unlike other neurotoxic agents which cause increase of intracellular calcium level, BPA decreased $[Ca^{2+}]_i$ dose-dependently in PC12 cells and cortical neuronal cells regardless of the calcium existence in buffer. BPA at greater concentrations than 100 $\mu\textrm{M}$ reduced cell viability significantly in both types of cells. BPA also suppressed L-glutamate (L-type channel activator, 30 mM) and trifluoperazine (calmodulin antagonist, 30 $\mu\textrm{M}$)-induced increase of $[Ca^{2+}]_i$. BPA further lowered caffeine (RYR activator, 100 $\mu\textrm{M}$)-decreased $[Ca^{2+}]_i$, but did not alter dantrolene (RYR inhibitor, 100 $\mu\textrm{M}$), heparin (IP3 inhibitor, 200 units/ml) and xestospongin C (IP3 inhibitor, 5 $\mu\textrm{M}$)-decreased $[Ca^{2+}]_i$. Cell viability was not directly related to intracellular calcium change by bisphenol A that alternation of intracellular calcium may not be a direct causal factor of BPA-induced neuronal cell death.