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Purpose: This study aims to describe and identify in-depth the breastfeeding experience of primiparas for more than one year using a phenomenological research method and to raise the understanding of the primiparas’ subjective experiences during the breastfeeding process to contribute to the improving and maintaining maternal and child health.Methods: This research was a phenomenological and qualitative study conducted on 5 primiparas who continued breastfeeding for more than 12 months. Data was collected through individual in-depth interviews from November 1, 2021 to March 20, 2022, and it was analyzed through the sixth stage of phenomenological research analysis presented by Colaizzi.Results: The study consisted of 10 theme clusters and 26 themes from meaningful statements about the breastfeeding success experiences of primiparas for more than one year. The 10 theme collections were "Planning breastfeeding during pregnancy," "Commitment to breastfeeding through information and education," "Challenges faced during breastfeeding," "Continuous hard work," "Continued conflict over breastfeeding amid confusion," "Connects with experts who lead the way," "Establishes one's own standards and direction," "Peace after hard work," "Solid and firm confidence," and "Deep-rooted beliefs brestfeeding serve as reminders."Conclusion: It is expected that this study will help to understand the individual difficulties and problems experienced by primiparas during breastfeeding, the importance of breastfeeding education, the role of medical personnel in the early stage of childbirth, social and environmental support, and the importance of an active window for information exchange. By identifying these factors, this study can serve as basic data for nursing practice to promote successful breastfeeding.

Aptamers are widely used as binders that interact with targets with high affinity or as inhibitors of the function of target molecules. However, they have also been used to modulate target protein function, which they achieve by activating the target or stabilizing its conformation. Here, we report a unique aptamer modulator of the insulin receptor (IR), IR-A62. Alone, IR-A62 acts as a biased agonist that preferentially induces Y1150 monophosphorylation of IR. However, when administered alongside insulin, IR-A62 shows variable binding cooperativity depending on the ligand concentration. At low concentrations, IR-A62 acts as a positive allosteric modulator (PAM) agonist that enhances insulin binding, but at high concentrations, it acts as a negative allosteric modulator (NAM) agonist that competes with insulin for IR. Moreover, the concentration of insulin affects the binding of IR-A62 to IR. Finally, the subcutaneous administration of IR-A62 to diabetic mice reduces blood glucose levels with a longer-lasting effect than insulin administration. These findings imply that aptamers can elicit various responses from receptors beyond those of a simple agonist or inhibitor. We expect further studies of IR-A62 to help reveal the mechanism of IR activation and greatly expand the range of therapeutic applications of aptamers.

Foodborne diseases (FBDs) remain a global public health concern. Climatic factors such as wind-chill temperature, rainfall, and relative humidity affect the incidence of several FBDs. This study was performed to analyze how the various factors of the climate influence the incidence and severity of FBDs. This study retrospectively analyzed the results of multiplex polymerase chain reaction (mPCR) tests for diarrhea-causing bacteria performed on 2300 fecal samples obtained from patients at Dankook University Hospital, Cheonan, from June 2010 to December 2019. The Clostridioides difficile toxin B infection rate positively correlated with the intensity of sunshine, and the content of particulate matter. The Campylobacter spp. infection rate positively correlated with wind-chill temperature and the content of particulate matter. The Escherichia coli O157:H7 infection rate positively correlated with relative humidity. These findings may explain the dynamics and risks of Clostridioides difficile toxin B, Clostridium perfringens, Campylobacter spp., and Escherichia coli O157:H7 infection. They may help predict interrelationships among climatic factors and standardize national environmental health policies. However, in-depth research with large-scale data, molecular biology, and epidemiology would be required going forward. Future research would also require objective indicators of the changes in the prevalence of FBD-causing microbial pathogens for the effective prevention and management of these bacterial infections.

Heavy metals such as lead (Pb) and cadmium (Cd) exist as particulate matter (PM) in the air and can cause biological damage to cells, animals, and humans. However, the mechanism underlying the toxic effects of heavy metals on nerve cells has not yet been completely identified. Glioma is the most common and fatal tumor in the central nervous system; the U87 human glioblastoma cell line is commonly used when researching brain cancer, including aggressive malignant gliomas. Therefore, in this study, cell viability, cytotoxicity, and interleukin-6 (IL-6) levels were analyzed to confirm the effect of Cd and Pb exposure on U87 cells. On confirming the absence of significant effects on cell viability at low concentrations of heavy metals, Cd and Pb exposure had no effect on lactic acid dehydrogenase (LDH) activity at the concentrations (1 μg/L, 30 μg/L, and 1 mg/L) used in this study, and there was a remarkable effect of Cd and Pb exposure on the inflammatory response of these cells. Our findings provide a basis for future research elucidating the effects of heavy metal exposure on cellular pathology. Systematic studies with higher heavy metal concentrations and precision are warranted to deepen our understanding of the relationship between heavy metal exposure and neuronal responses.

Rotavirus A is the most common cause of infectious diarrhea worldwide. This study aimed to retrospectively study and analyze 4009 stool samples that were tested for viruses causing diarrhea, using multiplex reverse transcription PCR at Dankook University Hospital between 2010 and 2019. Furthermore, we determined the correlation between these factors and various climatic factors, including wind-chill temperature, relative humidity, rate of sunshine, and particulate matter. Rotavirus A infections occurred frequently in February, March, and April on an annual basis. Furthermore, during the study, the detection rate was highest at 17.0% (n=61/359) in 2011. Based on an analysis of weather big data, patient age, and period-specific infection during the summer, when the wind-chill temperature and relative humidity were high, the Rotavirus A infection rate was very low. Relative humidity (p=0.020) and particulate matter (p=0.049) were associated with the average number of monthly cases of Rotavirus A infection. However, wind chill temperature (p=0.074) and rate of sunshine (p=0.993) were not associated with the average monthly distribution of Rotavirus A cases. These results indicate that Rotavirus A infection was correlated with relative humidity and particulate matter during the study period and further the current understanding of the distribution of Rotavirus A infections resulting from climatic factors and particulate matter. This could help establish climate-related health policies to reduce the incidence of diarrhea and guide the development of vaccines against Rotavirus A.

Studying relationships between meteorological conditions and respiratory virus infections may help interpret the causality of disease outbreaks and provide a better understanding of the seasonal distribution of viruses. Therefore, in this study, we analyzed the correlations between meteorological data and the trends of infection by human parainfluenza virus-1 (HPIV-1; also known as human respirovirus 1), human parainfluenza virus-2 (human orthorubulavirus 2), and human parainfluenza virus-3 (human respirovirus 3) using 9010 viral samples collected at Dankook University Hospital from January 1, 2012, to December 31, 2018. Infection frequency data were used to detect the seasonal patterns of HPIV-1, HPIV-2, and HPIV-3 infections, and these patterns were compared with local weather data over the same period. We performed descriptive statistical analysis, frequency analysis, t test, and binomial logistic regression analysis to examine the relationships of weather and particulate matter conditions with the incidence of HPIV-1, HPIV-2, and HPIV-3 infections. The highest average infection rate with one of these three viruses (88.17%) was found in children aged 1-9 years. Specifically, the infection rate of HPIV-1 was 91.9% in children aged 1-9 years, whereas that of HPIV-2 and HPIV-3 was 86.3%. HPIV infection exhibited a meaningful relationship with climatic factors, such as temperature, wind-chill temperature, and atmospheric pressure. Our results suggest that climate changes might affect the rate of infection by HPIV. These findings may help in predicting the effectiveness of preventive strategies of HPIV infection.

Purpose: The purpose of this study was to investigate psychological capital, sense of community and stress in relation to school adjustment and to identify factors influencing school adjustment in middle school students. Methods: This study population consisted of 150 middle school students. Data were analyzed using t-test, analysis of variance, Pearson correlation coefficient, and multiple regression analysis. Results: The mean score for school adjustment in middle school students was 3.68 out of 5 points. School adjustment explained 34% of the variance in sense of community (β=.39, p

In this paper, we propose a blind method to detect the mode of AMR speech data. This method provides a compression mode information by use of investing bit stream of unknown compressed data. Unlike uncompressed data, compressed data is inevitable in communication channels due to their efficient data size. However, the compressed data can lose their critical information such as the mode of AMR since the information is possibly delivered in a different channel or lost in a memory device. For these reasons, we propose an algorithm to detect the mode information of AMR when the mode information is missing. The algorithm consists of the processes of analyzing the compressed bitstreams and defining a specific group of bits to detect the mode information efficiently. To evaluate our method, we conduct experiments. Our experiments indicate that the proposed method provides sufficient performance to classify the mode information of AMR.

Most cameras use a color filter array (CFA) to capture color images. This array has only one color among red, green and blue per pixel and so, each pixel on the sensor lacks two color channels which can be reconstructed by a process called demosaicking. This paper proposes a demosaicking method for a plenoptic camera. Plenoptic raw images have a particular lenslet structure which must be formed the empty space, unlike conventional images. These vacant spaces exist because the lens is circular. The existing demosaicking methods degrade the image quality because it interpolates missing colors, including information on the empty space. Recently, a method using information about empty spaces has been studied. However, this method uses empty space information only for G-plane. To improve this method we applied information about empty space to R/B-plane too. Experimental results show that the performance of the proposed method is superior. As compared with the existing demosaicking algorithms, we show that our solution produces the best average demosaicking performance both objectively and subjectively.

This descriptive correlational study was to intended to analyze the relationship among emotional intelligence, academic self-efficacy and career decision level in high school girls. Data were collected from 241 high school girls in B and C city and analyzed using SPSS 20.0. The results are as follows: First, the mean score of emotional intelligence, academic self-efficacy and career decision level were above moderate. The levels of score were significant differences according to grade, school record, economic status, hoping career, and peer relationship. There were positive correlations among emotional intelligence, academic self-efficacy and career decision level. Therefore, it is necessary to consider emotional intelligence and academic self-efficacy for improving career decision level of high school girls.

Melton demonstrated a surprising disappearance using total internal reflection. When he put a Florence flask filled with marbles into a water tank and looked straight down from directly above the flask, he was only able to see marbles above a certain water level. When he added more water into the tank above the top line of the marbles, all of the marbles disappeared. He explained this phenomenon as due to a combination of both refraction and total internal reflection. Here, we wanted to develop this surprising idea to create more surprises. However, in our case, we only took the refraction effect from Melton’s idea to demonstrate our magic. This idea is supported by various perspectives. For instance, Viss and Sikkema demonstrated the critical angle without using total internal reflection, and James showed the novel optical properties of a submerged light bulb.

[Purpose] This study investigates the effects of task-oriented activities on hand function, cognitive function, and self-expression of the elderly with dementia, and then identify the influencing factors on self-expression in sub-factors of dependent variables. [Subjects and Methods] Forty elderly persons were divided into two groups: intervention group (n=20) and control group (n=20). The interventions were applied to the subjects 3 times a week, 50 minutes per each time, for a total of five weeks. We measured the jamar hand dynamometer test for grip strength, the jamar hydraulic pinch gauge test for prehension test, nine-hole pegboard test for coordination test, and Loewenstein Occupational Therapy Cognitive Assessment-Geriatric Population for cognitive function, and self-expression rating scale for self-expression test. [Results] The task-oriented activities promoted hand function, cognitive function (visual perception, spatial perception, visuomotor organization, attention & concentration) and self-expression of the elderly with early dementia, and the factors influencing the self-expression were cognitive function (visual perception) and hand function (coordination). The study showed that the task-oriented program enabled self-expression by improving hand function and cognitive function. [Conclusion] This study suggested that there should be provided the task-oriented program for prevention and treatment of the elderly with early dementia in the clinical settings and it was considered that results have a value as basic data that can be verified relationship of hand function, cognitive function, and self-expression.

BW723C86, a serotonin receptor 2B agonist, has been investigated as a potential therapeutic for various conditions such as anxiety, hyperphagia and hypertension. However, the functional role of BW723C86 against melanogenesis remains unclear. In this study, we investigate the effect of serotonin receptor 2B (5-HTR2B) agonist on melanogenesis and elucidate the mechanism involved. BW723C86 reduced melanin synthesis and intracellular tyrosinase activity in melan-A cells and normal human melanocytes. The expression of melanogenesis-related proteins (tyrosinase, TRP-1 and TRP-2) and microphthalmia-associated transcription factor (MITF) in melan-A cells decreased after BW723C86 treatment. The promoter activity of MITF was also reduced by BW723C86 treatment. The reduced level of MITF was associated with inhibition of protein kinase A (PKA) and cAMP response element-binding protein (CREB) activation by BW723C86 treatment. These results suggest that the serotonin agonist BW723C86 could be a potential therapeutic agent for skin hyperpigmentation disorders.

Aptamers are small synthetic oligonucleotides that bind to target proteins with high specificity and affinity. AS1411 is an aptamer that binds to nucleolin, which is overexpressed in the cytoplasm and occurs on the surface of cancer cells. We investigated the therapeutic potential of aptamers in hepatocellular carcinoma (HCC) by evaluating anti-tumor effects and confirming the affinity and specificity of AS1411- and modified AS1411-aptamers in HCC cells. Cell growth was assessed using the MTS assay, and cell death signaling was explored by immunoblot analysis. Fluorescence-activated cell sorting was performed to evaluate the affinity and specificity of AS1411-aptamers in SNU-761 HCC cells. We investigated the in vivo effects of the AS1411-aptamer using BALB/c nude mice in a subcutaneous xenograft model with SNU-761 cells. Treatment with a modified AS1411-aptamer significantly decreased in vitro (under normoxic [P = 0.035] and hypoxic [P = 0.018] conditions) and in vivo (under normoxic conditions, P = 0.041) HCC cell proliferation compared to control aptamers. AS1411- and control aptamers failed to control HCC cell proliferation. However, AS1411- and the modified AS1411-aptamer did not induce caspase activation. Decrease in cell growth by AS1411 or modified AS1411 was not prevented by caspase or necrosis inhibitors. In a microarray, AS1411 significantly enhanced galectin-14 expression. Suppression of HCC cell proliferation by the modified AS1411-aptamer was attenuated by galectin-14 siRNA transfection. Modified AS1411-aptamer suppressed HCC cell growth in vitro and in vivo by up-regulating galectin-14 expressions. Modified AS1411-aptamers may have therapeutic potential as a novel targeted therapy for HCC.

Objectives: This study was to identify the relationship between the Menopausal symptoms, Menopausal knowledge, Menopausal attitude and Menopausal management of the Middle Aged Women. Methods: The data was collected from three hundred and five Middle Aged Women whose children are studying in K university which is located in B metropolitan city. Results: The mean score of menopause symptoms, menopause knowledge, menopause attitude and menopause management among subjects were 41.08±8.78, 12.66±2.32, 58.67± 8.35 and 46.63±6.91 respectively. There was a positive relation between menopause symptoms and menopause knowledge (r=-.159, p=.006). But, between menopause symptoms and menopause attitude, there was a negative correlation (r=-.130, p=.023). This result showed a significant difference between menopause symptoms and the variables of age, education, job, physical health, and hobby. Also, it showed a significant correlation between menopause management and the variables of religion, income, satisfaction level with life, hobby, exercise, eating habits, eating pattern and regular medical check-up. Conclusion: The present work suggested that it is necessary to develop nursing intervention programs to increase the ability of menopause management through further research on influential factors in menopause symptoms and menopause management.

Anti-angiogenic therapeutics has been investigated extensively for the treatment of retinal and choroidal vascular diseases, and diabetic retinopathy. Anti-Flt1 peptide of GNQWFI is an antagonistic peptide for vascular endothelial growth factor receptor 1 (VEGFR1 or Flt1) inhibiting VEGFR1-mediated endothelial cell migration and tube formation. In this work, anti-Flt1 peptide (GGNQWFI) was chemically conjugated to tetra-n-butyl ammonium modified hyaluronate (HA-TBA) via amide bond formation in dimethyl sulfoxide (DMSO) using benzotriazol-1-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate (BOP). The resulting HA - GGNQWFI conjugate self-assembled to form micelle-like nanoparticles in aqueous solution, as confirmed and characterized by transmission electron microscopy (TEM). According to in vitro biological activity tests, HA - GGNQWFI conjugate exhibited a dose-dependent inhibition effect on the binding of Flt1-Fc to VEGF(165) coated on the well. Furthermore, anti-Flt1 peptide - HA conjugate effectively inhibited retinal choroidal neovascularization (CNV) in laser induced CNV model rats. The retinal vascular permeability and the deformation of retinal vascular structure were also significantly reduced in diabetic retinopathy model rats after treatment with anti-Flt1 peptide - HA conjugate. Pharmacokinetic analysis confirmed the increased mean residence time of anti-Flt1 peptide after conjugation to HA longer than 2 weeks.

Hyaluronic acid (HA) is a biodegradable, biocompatible, non-toxic, non-immunogenic and non-inflammatory linear polysaccharide, which has been used for various medical applications such as arthritis treatment, ocular surgery, tissue augmentation, and so on. In this review, the effect of chemical modification of HA on its distribution throughout the body was reported for target specific and long-acting delivery applications of protein, peptide, and nucleotide therapeutics. According to the real-time bio-imaging of HA derivatives using quantum dots (QDot), HA-QDot conjugates with 35mol% HA modification maintaining enough binding sites for HA receptors were mainly accumulated in the liver, while those with 68mol% HA modification losing much of HA characteristics were evenly distributed to the tissues in the body. The results are well matched with the fact that HA receptors are abundantly present in the liver with a high specificity to HA molecules. Accordingly, slightly modified HA derivatives were used for target specific intracellular delivery of nucleotide therapeutics and highly modified HA derivatives were used for long-acting conjugation of peptide and protein therapeutics. HA has been also used as a novel depot system in the forms of physically and chemically crosslinked hydrogels for various protein drug delivery. This review will give you a peer overview on novel HA derivatives and the latest advances in HA-based drug delivery systems of various biopharmaceuticals for further clinical development.

Agonistic and antagonistic peptides for formyl peptide receptor like 1 (FPRL1) receptor have been investigated as novel drug candidates for inflammatory diseases such as sepsis, asthma, and rheumatoid arthritis. In this work, a novel protocol for the synthesis of hyaluronic acid (HA)-peptide (CWRYMVm) conjugate for FPRL1 receptor was successfully developed for further clinical applications of peptide drugs. Aminoethyl methacrylated HA (HAAEMA) was synthesized by the coupling reaction of tetrabutyl ammonium salt of HA (HA-TBA) and AEMA using benzotriazol-1-yloxy-tris(dimethylamino) phosphonium hexafluorophosphate (BOP) in dimethyl sulfoxide (DMSO). Then, HA-AEMA was conjugated with CWRYMVm in water via Michael addition reaction between methacrylate group of HA-AEMA and thiol group in cysteine. The formation of HA-peptide conjugate was confirmed by 1H NMR and gel permeation chromatography (GPC). The average number of conjugated peptide molecules could be controlled from 5 to 23 per single HA chain. The HA-peptide conjugate showed serum stability longer than four days. In Vitro signal transduction activity of the HA-peptide conjugate for FPRL1 receptor was confirmed from the elevated levels of phospho-extracellular signal-regulated kinase (pERK) and calcium ion in FPRL1 overexpressing RBL-2H3 cells. The partially decreased biological activity of HA-peptide conjugates by the steric hindrance of HA was recovered after its degradation by hyaluronidase treatment.

A novel protocol to control the molecular degradation of hyaluronic acid (HA) hydrogels was successfully developed for tissue augmentation applications. HA has a different conformational structure in water and organic solvent, and the carboxyl group of HA is known to be the recognition site of hyaluronidase and HA receptors. Based on these findings, HA was chemically modified by grafting adipic acid dihydrazide (ADH) to the carboxyl group of HA in the water to prepare HA-ADH(WATER) and in the mixed solvent of water and ethanol to prepare degradation-controlled HA-ADH(WATER/ETHANOL). Three kinds of HA hydrogels were prepared by the crosslinking of HA-ADH(WATER) or HA-ADH(WATER/ETHANOL) with bis(sulfosuccinimidyl) suberate, and by the crosslinking of HA-OH with divinyl sulfone (DVS). In vitro and in vivo degradation tests showed that HA-DVS hydrogels were degraded most rapidly, followed by HA-ADH(WATER) hydrogels and HA-ADH(WATER/ETHANOL) hydrogels. There was no adverse effect during and after in vivo degradation tests. All of the HA hydrogel samples appeared to be biocompatible, according to the histological analysis with hematoxylin-eosin and Alcian blue.

A novel non-viral gene delivery system comprised of a DNA/PEI/Alginate (DPA) polyplex was prepared and assessedin vitro andin vivo. Coating the positively charged DNA/PEI (DP) complex with a polyanion resulted in a high level ofin vitro reporter gene transfection in the presence of 50 vol% serum due to the minimized cytotoxicity of PEI and the reduced nonspecific interactions with serum components. Among the tested anionic polymers, which included sodium alginate, poly(methacrylic acid) and poly(acrylic acid), the sodium alginate showed the highest gene transfection efficiency. The DPA polyplex also showed a reduced level of erythrocyte aggregation in target cells when compared with the DP complex. According toin vivo studies in which reporter genes encoding green fluorescent protein (GFP) and luciferase were used, injection of the DPA polyplex into tumor cells in six week old female C57/BL6 mice resulted in a much higher level of GFP expression and approximately 7 fold higher luciferase expression than treatment with the DP complex. Taken together, these results demonstrated that the anionic alginate coating of the DP complex contributed to efficient gene deliveryin vitro andin vivo.

In a previous study, we found that the phylogenetic analysis of partial rpoB sequences can be used effectively to phylogenetically differentiate Streptomyces spp. [B.J. Kim, C.J. Kim, J. Chun, Y.H. Koh, S.H. Lee, J.W. Hyun, C.Y. Cha, Y.H. Kook, Phylogenetic analysis of the genera Streptomyces and Kitasatospora based on partial RNA polymerase beta-subunit gene (rpoB) sequences, Int. J. Syst. Evol. Microbiol. 54 (2004) 593-598]. In the present study, we analyzed the partial rpoB gene sequences of 19 reference Streptomyces strains associated with potato scab. Furthermore, to empirically confirm the usefulness of rpoB gene analysis for the phylogenetic differentiation of Streptomyces spp., we applied the proposed system to 27 potato scab isolates obtained from the Korean provinces of Jeju-do and Kangwon-do. Phylogenetic relationships among these isolates using the devised rpoB gene-based methods were generally similar to those reported for 16S rRNA gene-based analysis. Isolates from potato scab lesion in Korea were also clearly differentiated into their phylogenetic groups by this method. In addition, the deduced RpoB amino acid sequences were also found to be useful for differentiating these strains. Our data demonstrate that the rpoB gene-based method can be used as a means of complementing other genetic methods such as 16S rRNA gene analysis or DNA-DNA hybridization to phylogenetically differentiate potato scab related Streptomyces spp.

Although Korea is a hepatitis B virus (HBV) endemic area, relatively few full-length genome sequences are available. In particular, no comparative analysis has been performed on the full-genome sequences of different HBV quasispecies from a single Korean patient. This report describes the full-length sequences of five HBV clones (two clones with shorter PCR amplicons and three clones with longer amplicons). Large deletions, that is, 685-bp, 487-bp, and 144-bp, that might interfere with the production of normal proteins were observed in four of five clones. Double mutations in the basal core promoter (BCP) region (T1762/A1764) were detected in two clones but no precore mutations (A1896) were detected in any of the five clones. These data support previous results that genotype C, in particular Korean clones of this genotype, is prone to mutations. Two independent mechanisms, namely, the deletions of long lengths and amino acid substitutions followed by BCP double mutations might contribute to the diversity of HBV quasispecies. Considering the importance of HBV quasispecies as HBV variant sources, the distribution of HBV quasispecies in mutation prone genotype C prevalent areas like Korea, should be monitored to improve the management of chronic HBV infections and to control HBV variants that arise due to the administration of vaccine or antiviral therapy.

A novel sustained release formulation of erythropoietin (EPO) was successfully developed using hyaluronic acid (HA) hydrogels crosslinked by Michael addition. Adipic acid dihydrazide grafted HA (HA-ADH) was prepared and then modified into methacrylated HA (HA-MA). (1)H NMR analysis showed that the degrees of HA-ADH and HA-MA modification were 69 and 29 mol%, respectively. Using the specific crosslinkers of dithiothreitol (DTT) and peptide linker, EPO was loaded during HA-MA hydrogel preparation by Michael addition chemistry between thiol and methacrylate groups. The amount of EPO recovered from both hydrogels after degradation with hyaluronidase SD (HAse SD) was about 90%. The crosslinking reaction with peptide linker (GCYKNRDCG) was faster than that with DTT. The gelation time was about 30 min for peptide linker and 180 min for DTT. In vitro release test of EPO from HA-MA hydrogel at 37 degrees C showed that EPO was released rapidly for 2 days and then slowly up to 7 days from HA-MA hydrogels. The released EPO appeared to be intact from the analysis with RP-HPLC. According to in vivo release test of EPO from HA-MA hydrogels crosslinked with the peptide linker in Sprague-Dawley (SD) rats, elevated plasma concentration of EPO was maintained up to 7 days. There was no adverse effect during and after the in vivo tests.