• Title/Summary/Keyword: Direct-Injection

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Biogenesis of Lysosome-related Organelle Mutant Silkworms by Direct Injection of a Cas9 Protein-guided RNA Complex into Bombyx mori Embryos (Cas9 단백질/ 가이드 RNA 복합체를 이용한 누에 BmBLOS 유전자 편집)

  • Kim, Kee Young;Yu, Jeong Hee;Kim, Su-Bae;Kim, Seong-Wan;Kim, Seong-Ryul;Choi, Kwang-Ho;Kim, Jong Gil;Park, Jong Woo
    • Journal of Life Science
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    • v.29 no.5
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    • pp.537-544
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    • 2019
  • Genome editing technology employing gene scissors has generated interest in molecular breeding in various fields, and the development of the third-generation gene scissors of the clustered, regularly interspaced short palindromic repeat (CRISPR) system has accelerated the field of molecular breeding through genome editing. In this study, we analyzed the possibility of silkworm molecular breeding using gene scissors by genomic and phenotypic analysis after editing the biogenesis of lysosome-related organelles (BmBLOS) gene of Bakokjam using the CRISPR/Cas9 system. Three types of guide RNAs (gRNA) were synthesized based on the BmBLOS gene sequence of Bakokjam. Complexes of the prepared gRNA and Cas9 protein were formed and introduced into Bombyx mori BM-N cells by electroporation. Analysis of the gene editing efficiency by T7 endonuclease I analysis revealed that the B4N gRNA showed the best efficiency. The silkworm genome was edited by microinjecting the Cas9/B4N gRNA complex into silkworm early embryos and raising the silkworms after hatching. The hatching rate was as low as 18%, but the incidence of mutation was over 40%. In addition, phenotypic changes were observed in about 70% of the G0 generation silkworms. Sequence analysis showed that the BmBLOS gene appeared to be a heterozygote carrying the wild-type and mutation in most individuals, and the genotype of the BmBLOS gene was also different in all individuals. These results suggest that although the possibility of silkworm molecular breeding using the CRISPR/Cas9 system would be very high, continued research on breeding and screening methods will be necessary to improve gene editing efficiency and to obtain homozygotes.

Arthroscopic Iliopsoas Tenotomy of Iliopsoas Impingement after Total Hip Arthroplasty (고관절 전치환술 후 발생한 장요건 충돌의 관절경하 장요건 절단술)

  • Huh, Soon Ho;Choi, Byeong Yeol;Han, Sang Roc;Chung, Woo Chull
    • Journal of the Korean Orthopaedic Association
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    • v.56 no.2
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    • pp.125-133
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    • 2021
  • Purpose: The clinical outcomes were investigated to determine if arthroscopic management is a useful method for 19 hips with iliopsoas tendon impingement (IPI) after total hip arthroplasty (THA). Materials and Methods: Eighteen patients (19 hips), who complained of groin pain and flexion pain that persisted after THA from September 2013 to December 2019, were the subjects of this investigation. The mean time to manifestation after THA was four months (range, 1-9 months) in patients of an average age of 60 years (range, 50-69 years). Thirteen out of 18 patients underwent THA using the direct anterior approach and five by the lateral approach. IPI was diagnosed by the medical history, physical examination, blood test, radiographic examination using X-ray and computed tomography, and topical injection therapy. All patients underwent arthroscopic treatment and a dynamic arthroscopic physical examination after exposure to the iliopsoas tendon revealed impingement. Tenotomy was then performed on the muscle portion through the total tendon portion. Symptoms and pain levels of preoperative, postoperative and follow-up period were investigated and compared. Results: The Western Ontario and McMaster Universities Osteoarthritis Index score decreased from an average of 58.4 (range, 40-88) before surgery to an average of 35.0 (range, 15-76) after surgery. Similarly, the visual analogue scale decreased from an average of 4.0 (range, 2-6) before surgery to an average of 1.4 (range, 0-4) after surgery. Sixteen patients (88.9%) showed pain relief and improvement in the straight leg raise test, and two patients showed postoperative muscle weakness and sustained pain. In the follow-up period, muscle weakness improved. One patient underwent arthroscopic iliopsoas tenotomy at the lesser trochanteric level but the symptoms persisted. The clinical symptoms were improved after one more tenotomy at the joint level. Conclusion: Arthroscopic iliopsoas tenotomy performed in patients with IPI after THA showed good clinical results.

A Study on the Expressed Desire at Discharge of Patients to Use Home Nursing and Affecting Factors of the Desire (퇴원환자의 가정간호 이용의사와 관련 요인)

  • Lee, Ji-Hyun;Lee, Young-Eun;Lee, Myung-Hwa;Sohn, Sue-Kyung
    • The Korean Journal of Rehabilitation Nursing
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    • v.2 no.2
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    • pp.257-270
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    • 1999
  • The purpose of this study is to investigate factors related to the intent of using home nursing of chronic disease patients who got out of a university hospital. For the purpose, the study selected 153 patients who were hospitalized and left K university hospital with diagnoses of cancer, hypertension, diabetes and cerebral vascular accident and ordered to be discharged and performed interviews with them and surveys on their medical records to obtain the following results. For this study a direct-interview survey and medical record review was conducted from June 28 to Aug. 30, 1998. The frequency and mean values were computed to find the characteristics of the study subjects, and $X^2$-test, t-test, factor analysis and multiple logistic regession analysis were applied for the analysis of the data. The following results were obtained. 1) When characteristics of the subjects were examined, men and women occupied for 58.8% and 41.2%, respectively. The subjects were 41.3 years old in aver age and had the monthly aver age earning of 0.99 million won or below, which was the most out of the total subjects at 34.6%. Among the total, 87.6% resided in cities and 12.4 in counties. The most left the hospital with diagnosis of cancer at 51.6%, followed by hyper tension at 24.2%, diabetes at 13.7% and cerebral vascular accident at 7.2%. 2) 93.5% of the selected patients had the intent of using home nursing and 6.5%, didn't. Among those patients having the intent, 85.6% had the intent of paying for home nursing and 14.4%, didn't. The subjects expected that the nursing would be paid 9,143 won in aver age and 47.7% of them preferred national authorities as the main servers. 86.3% of the subjects thought that home nursing business had the main advantage of making it possible to learn nursing methods at home and thereby contributing to improving the ability of patients and their facilities to solve health problems. 3) Relations between the intent of use and characteristics of the subjects such as demography-related social, home environment, disease and physical function characteristics did not show statistically significant differences among one another. Compared to those who had no intent of using home nursing, the group having the intent had more cases of male patients, the age of 39 or below, residence in cities, 5 family member s or more, no existence of home nursing servers, leaving the hospital from a non-hospitalized building, disease development for five months or below, hospitalization for ten days or more, non-hospitalization with in the recent one month, two times or over of hospitalization, leaving the hospital with no demand of special treatment, operation underwent, poor results of treatment, leaving the hospital with demand of rehabilitation services, physical disablement and high evaluation point of daily life. 4) Among those patients having the intent of using home nursing, 47.6% demanded technical nursing and 55.9%, supportive nursing. As technical nursing,' inject into a blood vessel ' and 'treat pustule and teach basic prevention methods occupied for 57.4%, respectively, topping the list. Among demands of supportive nursing, 'observe patients 'status and refer them to hospitals or community resources as available, if necessary' was the most with percent age point of 59.5. Regarding the intent of paying for home nursing, 39.2% of those patients wishing to use the nursing responded paying for technical services and 20.2, supportive services. In detail, 70.0% wanted to pay for a service stated as 'inject into a blood vessel', highest among the former services and 30.7%, a service referred to as 'teaching exercises needed to make the body of patients move', highest among the latter. When this was analyzed in terms of a relation between the need(the need for home nursing) and the demand(the intent of paying for home nursing), The rate of the need to the demand was found two or three times higher in technical nursing(0.82) than in supportive nursing(0.35). In aspects of tech ical nursing, muscle injection(1.26, the 1st rank) was highest in the rate while among aspects of supportive nursing, a service referred to as 'teach exercises needed for making patients move their bodies normally'(0.58, the 1st rank). 5) factors I(satisfaction with hospital services), II(recognition of disease state), III(economy) and IV(period of disease) occupied for 34.4, 13.8, 11.9 and 9.2 percents, respectively among factors related to the intent by the subjects of using home nursing, totaled 59.3%. In conclusion, most of chronic disease patients have the intent of using hospital-based home nursing and satisfaction with hospital services is a factor affecting the intent most. Thus a post-management system is needed to continue providing health management to those patients after they leave the hospital. Further, supportive services should be provided in order that those who are satisfied with hospital services return to their community and live their in dependent lives. Based on these results, the researcher would make the following recommendation. 1) Because home nursing becomes more and more needed due to a sharp increase in chronic disease patients and elderly people, related rules and regulations should be made and implemented. 2) Hospital nurses specializing in home nursing should be cultivated.

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