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Validation of Stem-loop RT-qPCR Method on the Pharmacokinetic Analysis of siRNA Therapeutics (Stem-loop RT-qPCR 분석법을 이용한 siRNA 치료제의 생체시료 분석법 검증 및 약물 동태학적 분석)

  • Kim, Hye Jeong;Kim, Taek Min;Kim, Hong Joong;Jung, Hun Soon;Lee, Seung Ho
    • Journal of Life Science
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    • v.29 no.6
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    • pp.653-661
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    • 2019
  • The first small interfering RNA (siRNA) therapeutics have recently been approved by the Food and Drug Administration in the U.S., and the demand for a new RNA therapeutics bioanalysis method-which is essential for pharmacokinetics, including the absorption, distribution, metabolism, and excretion of siRNA therapeutics-is rapidly increasing. The stem-loop real-time qPCR (RT-qPCR) assay is a useful molecular technique for the identification and quantification of small RNA (e.g., micro RNA and siRNA) and can be applied for the bioanalysis of siRNA therapeutics. When the anti-HPV E6/E7 siRNA therapeutic was used in preclinical trials, the established stem-loop RT-qPCR assay was validated. The limit of detection was sensitive up to 10 fM and the lower limit of quantification up to 100 fM. In fact, the reliability of the established test method was further validated in three intra assays. Here, the correlation coefficient of $R^2$>0.99, the slope of -3.10 ~ -3.40, and the recovery rate within ${\pm}20%$ of the siRNA standard curve confirm its excellent robustness. Finally, the circulation profiles of siRNAs were demonstrated in rat serum, and the pharmacokinetic properties of the anti-HPV E6/E7 siRNA therapeutic were characterized using a stem-loop RT-qPCR assay. Therefore, the stemloop RT-qPCR assay enables accurate, precise, and sensitive siRNA duplex quantification and is suitable for the quantification of small RNA therapeutics using small volumes of biological samples.

Questionnaire survey for the clinical trial participants who experienced both digital and conventional impression (디지털 인상법과 종래인상법을 동시에 경험한 임상시험자를 대상으로 한 설문지분석)

  • Yang, Eunbee;Kim, Bongju;Lee, Jun Jae;Lee, Seung-Pyo;Lim, Young-Jun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.34 no.4
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    • pp.270-279
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    • 2018
  • Purpose: The aim of this study was to assess the patients' perception, acceptance, and preference of the difference between a conventional impression and digital impression through questionnaire survey. Materials and Methods: Thirteen (6 male, 7 female) subjects who experienced both digital and conventional impression at the same day were enrolled in this study. Conventional impression were taken with polyvinylsiloxane and digital impression were performed using a newly developed intra-oral scanner. Immediately after the two impressions were made, a survey was conducted with the standardized questionnaires consisting of the following three categories; 1) general dental treatment 2) satisfaction of conventional impression 3) satisfaction of digital impression. The perceived source of satisfaction was evaluated using Likert scale. The distribution of the answers was assessed by percentages and statistical analyses were performed with the paired t-test, and P < 0.05 was considered significant. Results: There were significant differences of the overall satisfaction between two impression methods (P < 0.05). Digital impression showed high satisfaction in less shortness of breath and odor to participants compared to conventional impression. The use of an oral scanner resulted in a discomfort of TMJ due to prolonged mouth opening and in lower score of the scanner tip size. Conclusion: It was confirmed that the preference for the digital impression using intraoral scanner is higher than the conventional impression. Most survey participants said they would recommend the digital impression to others and said they preferred it for future prosthetic treatment.

The Optimization and Verification of an Analytical Method for Sodium Iron Chlorophyllin in Foods Using HPLC and LC/MS (식품 중 철클로로필린나트륨의 HPLC 및 LC/MS 최적 분석법과 타당성 검증)

  • Chong, Hee Sun;Park, Yeong Ju;Kim, Eun Gyeom;Park, Yea Lim;Kim, Jin Mi;Yamaguchi, Tokutaro;Lee, Chan;Suh, Hee-Jae
    • Journal of Food Hygiene and Safety
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    • v.34 no.2
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    • pp.148-157
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    • 2019
  • An optimized analytical method for sodium iron chloriphyllin in foods was established and verified by using high performance liquid chromatography with attached diode array detection. An Inertsil ODS-2 column and methanol-water (80:20 containing 1% acetate) as a mobile phase were employed. The limit of detection and quantitation of sodium iron chloriphyllin were 0.1 and 0.3 mg/kg, respectively, and the linearity of calibration curve was excellent ($R^2=0.9999$). The accuracy and precision were 93.9~104.95% and 2.0~7.7% in both inter-day and intra-day tests. Recoveries for candy and salad dressing were ranged between 93 and 104% (relative standard deviation, (RSD) 0.3~4.3%), and between 83 and 115% (RSD 1.2~2.0%), respectively. Liquid chromatography mass spectrometry was used to verify the main components of sodium iron chlorophyllin which were Fe-isochlorin e4 and Fe-chlorin e4.

Survival in clinical stage I endometrial cancer with single vs. multiple positive pelvic nodes: results of a multi-institutional Italian study

  • Uccella, Stefano;Falcone, Francesca;Greggi, Stefano;Fanfani, Francesco;De Iaco, Pierandrea;Corrado, Giacomo;Ceccaroni, Marcello;Mandato, Vincenzo Dario;Bogliolo, Stefano;Casarin, Jvan;Monterossi, Giorgia;Pinelli, Ciro;Mangili, Giorgia;Cormio, Gennaro;Roviglione, Giovanni;Bergamini, Alice;Pesci, Anna;Frigerio, Luigi;Uccella, Silvia;Vizza, Enrico;Scambia, Giovanni;Ghezzi, Fabio
    • Journal of Gynecologic Oncology
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    • v.29 no.6
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    • pp.100.1-100.13
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    • 2018
  • Objective: To investigate survival outcomes in endometrioid endometrial cancer (EEC) patients with single vs. multiple positive pelvic lymph nodes. Methods: We performed a retrospective evaluation of all consecutive patients with histologically proven International Federation of Gynecology and Obstetrics (FIGO) stage IIIC1 EEC who underwent primary surgical treatment between 2004 and 2014 at seven Italian gynecologic oncology referral centers. Patients with pre- or intra-operative evidence of extra-uterine disease (including the presence of bulky nodes) and patients with stage IIIC2 disease were excluded, in order to obtain a homogeneous population. Results: Overall 140 patients met the inclusion criteria. The presence of >1 metastatic pelvic node was significantly associated with an increased risk of recurrence and mortality, compared to only 1 metastatic node, at both univariate (recurrence: hazard ratio [HR]=2.19; 95% confidence interval [CI]=1.2-3.99; p=0.01; mortality: HR=2.8; 95% CI=1.24-6.29; p=0.01) and multivariable analysis (recurrence: HR=1.91; 95% CI=1.02-3.56; p=0.04; mortality: HR=2.62; 95% CI=1.13-6.05; p=0.02) and it was the only independent predictor of prognosis in this subset of patients. Disease-free survival (DFS) and disease-specific survival (DSS) were significantly longer in patients with only 1 metastatic node compared to those with more than 1 metastatic node (p=0.008 and 0.009, respectively). Conclusion: The presence of multiple metastatic nodes in stage IIIC1 EEC represents an independent predictor of worse survival, compared to only one positive node. Our data suggest that EEC patients may be categorized according to the number of positive nodes.

Performance of Investment Strategy using Investor-specific Transaction Information and Machine Learning (투자자별 거래정보와 머신러닝을 활용한 투자전략의 성과)

  • Kim, Kyung Mock;Kim, Sun Woong;Choi, Heung Sik
    • Journal of Intelligence and Information Systems
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    • v.27 no.1
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    • pp.65-82
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    • 2021
  • Stock market investors are generally split into foreign investors, institutional investors, and individual investors. Compared to individual investor groups, professional investor groups such as foreign investors have an advantage in information and financial power and, as a result, foreign investors are known to show good investment performance among market participants. The purpose of this study is to propose an investment strategy that combines investor-specific transaction information and machine learning, and to analyze the portfolio investment performance of the proposed model using actual stock price and investor-specific transaction data. The Korea Exchange offers daily information on the volume of purchase and sale of each investor to securities firms. We developed a data collection program in C# programming language using an API provided by Daishin Securities Cybosplus, and collected 151 out of 200 KOSPI stocks with daily opening price, closing price and investor-specific net purchase data from January 2, 2007 to July 31, 2017. The self-organizing map model is an artificial neural network that performs clustering by unsupervised learning and has been introduced by Teuvo Kohonen since 1984. We implement competition among intra-surface artificial neurons, and all connections are non-recursive artificial neural networks that go from bottom to top. It can also be expanded to multiple layers, although many fault layers are commonly used. Linear functions are used by active functions of artificial nerve cells, and learning rules use Instar rules as well as general competitive learning. The core of the backpropagation model is the model that performs classification by supervised learning as an artificial neural network. We grouped and transformed investor-specific transaction volume data to learn backpropagation models through the self-organizing map model of artificial neural networks. As a result of the estimation of verification data through training, the portfolios were rebalanced monthly. For performance analysis, a passive portfolio was designated and the KOSPI 200 and KOSPI index returns for proxies on market returns were also obtained. Performance analysis was conducted using the equally-weighted portfolio return, compound interest rate, annual return, Maximum Draw Down, standard deviation, and Sharpe Ratio. Buy and hold returns of the top 10 market capitalization stocks are designated as a benchmark. Buy and hold strategy is the best strategy under the efficient market hypothesis. The prediction rate of learning data using backpropagation model was significantly high at 96.61%, while the prediction rate of verification data was also relatively high in the results of the 57.1% verification data. The performance evaluation of self-organizing map grouping can be determined as a result of a backpropagation model. This is because if the grouping results of the self-organizing map model had been poor, the learning results of the backpropagation model would have been poor. In this way, the performance assessment of machine learning is judged to be better learned than previous studies. Our portfolio doubled the return on the benchmark and performed better than the market returns on the KOSPI and KOSPI 200 indexes. In contrast to the benchmark, the MDD and standard deviation for portfolio risk indicators also showed better results. The Sharpe Ratio performed higher than benchmarks and stock market indexes. Through this, we presented the direction of portfolio composition program using machine learning and investor-specific transaction information and showed that it can be used to develop programs for real stock investment. The return is the result of monthly portfolio composition and asset rebalancing to the same proportion. Better outcomes are predicted when forming a monthly portfolio if the system is enforced by rebalancing the suggested stocks continuously without selling and re-buying it. Therefore, real transactions appear to be relevant.

Tranexamic Acid Reduces Postoperative Blood Loss in Reverse Total Shoulder Arthroplasty (역행성 견관절 전치환술에서 트라넥삼산의 출혈 및 수혈 감소 효과)

  • Park, Kee Young;Kim, In Bo;Kim, Eun Yeol;Lee, Kwang Suk
    • Journal of the Korean Orthopaedic Association
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    • v.56 no.5
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    • pp.391-397
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    • 2021
  • Purpose: Tranexamic acid (TXA) can reduce perioperative blood loss and the frequency of blood transfusions in lower extremity surgery. On the other hand, the effects of TXA on reverse total shoulder arthroplasty (rTSA) remain undetermined. This study evaluated the efficacy of TXA on perioperative blood loss, transfusion requirements, and the change in the hemotologic index. Materials and Methods: This study evaluated patients who underwent rTSA from September 2009 to July 2020. The patients were classified into two groups. The TXA group were administered TXA intravenously and topical TXA during surgery. The non-TXA group was not administered TXA. The quantity of hemovac drainage, which represented the postoperative blood loss, transfusion requirements, and postoperative change in hemoglobin and hematocrit level, were recorded. Results: The TXA and non-TXA groups consisted of 93 and 84 patients, respectively. The preoperative demographics showed no significant differences in age (72.0±7.0 vs. 71.5±5.8, p=0.656), sex (male:female, 28:65 vs. 23:61, p=0.689) and the prevalence of hypertension and diabetes (hypertension:diabetes:both, 36:3:13 vs. 32:3:8, p=0.806) between the two groups. There were significant differences in the requirements of transfusion (0 vs. 9, p=0.001), hemovac drainage at the 1st (98.8±61.2 ml vs. 162.7±98.8 ml, p<0.001), the 2nd postoperative day (73.8±48.4 ml vs. 91.5±54.5 ml, p=0.024), hemoglobin level at the 1st (11.7±1.2 g/dl vs. 11.2±1.4 g/dl, p=0.048), 3rd (10.9±1.2 g/dl vs. 10.2±1.2 g/dl, p<0.001), and 6th (11.2±1.3 g/dl vs. 10.7±1.3 g/dl, p=0.020) postoperative day, and the hematocrit level at the 1st (35.0%±3.6% vs. 32.5%±3.8%, p=0.001), 3rd (32.3%±5.0% vs. 29.8%±3.6%, p<0.001), and 6th (33.5%±3.8% vs. 31.5%±3.7%, p<0.001) postoperative day between the two groups. Conclusion: Intravenous and topical intra-articular TXA can reduce the transfusion requirement and blood loss in rTSA.

Outcome of Conservative Treatment of the Zone I, II 5th Metatarsal Base Fracture under Early Weight-Bearing (제5 중족골 제1, 2 구역 골절의 조기 체중부하의 비수술적 치료 결과)

  • Gwak, Heui-Chul;Park, Dae-Hyun;Kim, Jung-Han;Lee, Chang-Rack;Kwon, Yong-Uk;Kim, Dong-Seok
    • Journal of the Korean Orthopaedic Association
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    • v.56 no.2
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    • pp.150-156
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    • 2021
  • Purpose: To determine how the location, displacement, intra-articular involvement, comminution of a 5th metatarsal base fracture affect results of early weight-bearing treatment. Materials and Methods: From January 2013 to July 2017, 34 cases of 34 patients diagnosed with a fracture of the zone I and II 5th metatarsal base were enrolled. The mean follow-up period was 13 months (6-15 months). One patient was excluded as a refracture during the follow-up period, and 33 patients underwent conservative treatment. Anteroposterior, lateral, and simple oblique radiography and computed tomography of the foot were performed to evaluate the location and displacement of the fracture, the degree of joint involvement, and comminution. In all 33 patients, a short leg cast or boot brace was selected immediately after the injury, tolerable weight bearing was allowed. If the pain disappeared, full weight bearing was performed after wearing a plain shoe or postoperative shoe. As a clinical result, the American Orthopedic Foot and Ankle Society (AOFAS) score was evaluated at the final follow-up. During outpatient follow-up, a simple radiograph of the foot was taken to confirm the time of radiological bone union and return to work. Results: Nine males and 24 females, with an average age of 48.7 years, were enrolled in the study. Twenty-four patients had zone I fractures, and nine patients had zone II fractures. Twenty-two out of 33 patients had a fracture displacement of 2 mm or more. Nine and five patients had joint involvement and comminution, respectively. There was a statistically significant return to work from zone I to zone II. The AOFAS score was excellent at the final follow-up and there was no significant difference. When classifying and comparing the degree of fracture displacement, joint involvement, and comminution, there were no significant differences in the radiological union time and return to work. In all cases, satisfactory results were obtained at the final follow-up. Conclusion: Satisfactory clinical results can be obtained by allowing early weight-bearing regardless of the fracture location, displacement, joint involvement, or comminution in zone I and II 5th metatarsal base fractures.

The Validation of a Commercial Testosterone RIA Test Kits (Testosterone RIA 검사 kit 별 유효성 비교평가)

  • Ryu, Hwa-jin;Shin, Seon-yeong;Cho, Seong-uk
    • The Korean Journal of Nuclear Medicine Technology
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    • v.26 no.1
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    • pp.38-41
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    • 2022
  • Purpose Testosterone is a steroid hormone synthesized by the Leydig cells of the testes in men, and by the adrenal cortex and ovaries in women. Testosterone production is regulated by luteinzing hormone secreted by the anterior pituitary gland. In this experiment, the effectiveness of testosterone radioimmunoassay (RIA) kits produced by three companies was evaluated and compared in case the production of testosterone kits was stopped or supply problems occurred. Materials and Methods In October 2021, samples were collected from the patients (n=49) who requested the testosterone RIA test. The experiment was conducted by dividing the patient's sample into low concentration (1.0 ng/mL or less), medium concentration (2.0-4.0 ng/mL) and high concentration (6.0 ng/mL or more). The Testosterone RIA test compared and evaluated the validity of Company A kits used in this hospital and those of Company B and C used in other hospitals. The precision, sensitivity, recovery, linearity and correlation were evaluated for each kit. The testosterone RIA test was carried out in accordance with the insert kit manual for each manufacturer. Results As a result of measuring the precision of the intra assay, the Coefficient of Variation (CV) value of the company A kit was high at 11.4% only in the low concentration sample, and in the case of the company B and C kits, the CV value was less than 10% at low, medium, and high concentrations. In the inter-assay precision measurement, the CV value was less than 15% in both A and C kits, but in the case of the B kit, the CV value exceeded 15% at low and medium concentrations. Sensitivity was 0.13 ng/mL for company A, 0.01 ng/mL for company B, and 0.01 ng/mL for company C, and the linearity of all three kits showed excellent linearity. In the case of recovery rate, all of the A, B, and C company kits showed results that were out of 90-110%. In the case of correlation test, when compared with the company A kit currently use in here, the correlation coefficient (R2) value for the company B kit was 0.9508, and for the company C kit was 0.9352 Conclusion As a result, there was a slight difference in precision at the low concentration sample. The correlation test showed an excellent correlation coefficient. However, it was difficult to secure samples of various concentrations because there were not many tests of testosterone requested at this hospital. So, additional experiments should be carried out by acquiring samples of various concentrations on each laboratory later.

Initial Evaluation using Geochemical Data to infer Tectonic Setting of Mt. Baekdu/Changbaishan Volcano (백두산 화산의 지체구조 추론을 위한 지구화학적 데이터를 이용한 기초 평가)

  • Yun, Sung-Hyo;Chang, Cheolwoo;Pan, Bo
    • Journal of the Korean earth science society
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    • v.43 no.1
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    • pp.128-139
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    • 2022
  • This study aimed to investigate the tectonic setting of the volcanic edifice at Mt. Baekdu by analyzing petrochemical characteristics of Holocene felsic volcanic rocks distributed in the Baekdusan stratovolcano edifice and summit of the Cheonji caldera rim, as well as Pleistocene mafic rocks of the Gaema lava plateau and Changbaishan shield volcano edifice. During the early eruption phases, mafic eruption materials, with composition ranging from alkali basalt to trachybasalt, or from subalkaline (tholeiitic) basalt to basaltic andesite formed the Gaema lava plateau and Changbaishan shield volcanic edifice, whereas the Baekdusan stratovolcano edifice and Holocene tephra deposits near the summit of the Cheonji caldera comprises trachytic and rhyolitic compositions. Analysis results revealed bimodal compositions with a lack of 54-62 SiO2, between the felsic and mafic volcanic rocks. This suggested that magmatic processes occurred at the locations of extensional tectonic settings in the crust. Mafic volcanic rocks were plotted in the field of within-plate volcanic zones or between within-plate alkaline and tholeiite zones on the tectonic discrimination diagram, and it was in good agreement with the results of the TAS diagram. Felsic volcanic rocks were plotted in the field of within-plate granite tectonic settings on discrimination diagrams of granitic rocks. None of the results were plotted in the field of arc islands or continental margin arcs. The primitive mantle-normalized spider diagram did not show negative (-) anomalies of Nb and Ti, which are distinctive characteristics of subduction-related volcanic rocks, but exhibited similar patterns of ocean island basalt. Trace element compositions showed no evidence of, magmatic processes related to subduction zones, indicating that the magmatic processes forming the Baekdusan volcanic field occurred in an intraplate environment. The distribution of shallow earthquakes in this region supports the results. The volcanic rocks of the Baekdusan volcanic field are interpreted as the result of intraplate volcanism originating from the upwelling of mantle material during the Cenozoic era.

Clinical Outcomes of Ultrasound-Guided Transmeniscal Injection in Medial Compartment Knee Osteoarthritis (무릎 내측 구획 골관절염에서 초음파 유도하 경 반월상 연골 주사의 임상적 효과)

  • Jung, Eui Yub;Wang, Joon Ho;Lee, Eui-Sub;Lee, Sung-Sahn;So, Sang-Yeon
    • Journal of the Korean Orthopaedic Association
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    • v.55 no.5
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    • pp.418-425
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    • 2020
  • Purpose: The purpose of this study was to introduce the ultrasound-guided transmeniscal injection in medial compartment knee osteoarthritis and analyze the clinical outcomes. Materials and Methods: The electronic medical records of 36 patients with medial compartment knee osteoarthritis who were treated with an ultrasound-guided transmeniscal injection from March 2019 to July 2019 were accessed for this retrospective review. Using an ultrasound guided spinal needle, the patients received an intra-articular steroid injection at the medial compartment of the knee. A physical examination was conducted at the initial visit (pre-injection), and at one week, four weeks, and eight weeks after the injection. The numeric pain rating scale (NRS), Lequesne index, and Western Ontario and McMaster Universities Osteoarthritis (WOMAC) score were measured at each visit and analyzed over time. The percentage change of the patients who revealed substantial improvement was analyzed. The NRS, Lequesne index, and percentage of patients, who revealed substantial improvement over time classified by osteoarthritis grade, were analyzed. Results: The NRS and Lequesne index decreased at one week, four weeks, and eight weeks after the injection compared to the initial baseline, and the pain-relief effect continued without change until eight weeks. The percentage of patients who showed substantial improvement at one, four, and eight weeks was 50.0%, 47.2%, and 52.8%, respectively. The WOMAC scores decreased at one, four, and eight weeks compared to the initial baseline, and the decrease was continued without any difference until eight weeks. The percentage of patients with osteoarthritis stage 1 or 2 who revealed more than substantial improvement was significantly higher at one, four, and eight weeks than those with osteoarthritis stages 3 or 4 (p<0.05). Conclusion: In patients with medial compartment knee osteoarthritis, the pain reduction and functional improvement persisted for at least eight weeks after the ultrasound-guided transmeniscal injection at the medial compartment. In particular, patients with medial compartment osteoarthritis stage 1 or 2 showed more effective pain reduction.