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In Vitro Culture of the Isolated Mouse Preantral Follicles: Effect of Different Types of FSH and Vitrification (생쥐 Preantral 난포의 체외배양: FSH의 종류와 농도 및 초자화 냉동보존의 영향)

  • Lee, Sook-Hyun;Shin, Chang-Sook;Chung, Hyung-Min;Ko, Jung-Jae;Cha, Kwang-Yul;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.4
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    • pp.387-392
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    • 2000
  • Objectives: 1) To compare the efficacy of urofollitropin (Follimon) to that of recombinant human FSH (rhFSH) on the growth and maturation of mouse early preantral follicles in vitro, and 2) effect of vitrification on the growth and maturation of preantral follicles and oocytes . Methods: Isolated early preantra1 follicles (100-130 ${\mu}m$ diameter) were cultured for 12 days in 20 ${\mu}l$ ${\alpha}$-MEM media drop under the mineral oil. Follimon or rhFSH was added to the culture medium at various concentrations (0, 10, 100, and 1000 mIU/ml). Results: With Follimon, the dose of 10 mIU/ml showed the best follicle survival, growth, and MIl rate of oocyte than the other concentrations. Whereas the optimal dose of rhFSH was 100 mIU/ml. Despite the different optimal doses, the efficacy of two different FSHs on the follicle growth and maturation was similar. Isolated mouse preantral follicles were cryopreserved by vitrification and cultured in vitro for 12 days with 100 mIU/ml rhFSH. Despite the decreased follicular survival rate after thawing, the follicular growth and maturation rate of its oocyte were comparable to those of the fresh follicle. Conclusion: Results from the present study revealed that 1) the optimal doses of Follimon and rhFSH for in-vitro culture of mouse follicles are different, and 2) the frozen-thawed follicles develop normally after vitrification.

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Phase II Clinical Study on the GEMOX Regimen as Second-line Therapy for Advanced Ovarian Cancer

  • Yuan, Shao-Fei;Zhang, Lian-Ping;Zhu, Lin-Jia;Chen, Wen-Jun;Zheng, Wei-E;Xiong, Jian-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3949-3953
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    • 2013
  • Aim: To investigate the effectiveness and adverse effects of gemcitabine by fixed-dose rate infusion plus oxaliplatin (GEMOX regimen) as second-line therapy for advanced ovarian cancer. Methods: 64 patients with advanced ovarian cancer were divided into an experimental group (44 cases) and a control group (20 cases). The experimental group was treated with continuous intravenous infusion of gemcitabine at 1000 $mg/m^2$ with a fixed-dose rate of 10 $mg/m^2/min$, on days 1 and 8 and oxaliplatin at 100 $mg/m^2$ on day 1, IVGTT, repeated every 3 weeks. The control group was treated with intravenous infusion of gemcitabine at 1000 $mg/m^2$ within 30 min on days 1 and and oxaliplatin at 100 $mg/m^2$ on day 1, IVGTT, again repeated every 3 weeks. CT scans or MRI were used for review every 1-2 cycles. Results: The effective rate in the experimental group was significantly high than control group (43.2% vs 35.0%; P < 0.05), with no obvious difference of hematologic or non-hematologic toxicity between the two groups (P > 0.05). Conclusion: GEMOX regimen is very effective to treat advanced ovarian cancer, with low toxicity, good tolerance and improved life quality in patients.

Laparoscopic Distal Gastrectomy for Gastric Cancer in Morbidly Obese Patients in South Korea

  • Jung, Ji Hoon;Ryu, Seong Yeop;Jung, Mi Ran;Park, Young Kyu;Jeong, Oh
    • Journal of Gastric Cancer
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    • v.14 no.3
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    • pp.187-195
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    • 2014
  • Purpose: Laparoscopic gastrectomy in obese patients has been investigated in several studies, but its feasibility has rarely been examined in morbidly obese patients, such as in those with a body mass index (BMI) of ${\geq}30kg/m^2$. The present study aimed to evaluate the technical feasibility and safety of laparoscopic gastrectomy in morbidly obese patients with gastric cancer. Materials and Methods: A total of 1,512 gastric cancer patients who underwent laparoscopic distal gastrectomy (LDG) were divided into three groups: normal (BMI< $25kg/m^2$, n=996), obese (BMI $25{\sim}30kg/m^2$, n=471), and morbidly obese ($BMI{\geq}30kg/m^2$, n=45). Short-term surgical outcomes, including the course of hospitalization and postoperative complications, were compared between the three groups. Results: The morbidly obese group had a significantly longer operating time (240 minutes vs. 204 minutes, P=0.010) than the normal group, but no significant differences were found between the groups with respect to intraoperative blood loss or other complications. In the morbidly obese group, the postoperative morbidity and mortality rates were 13.3% and 0%, respectively, and the mean length of hospital stay was 8.2 days, which were not significantly different from those in the normal group. Subgroup analysis showed that postoperative complication rates were not high in morbidly obese patients, independent of the type of anastomosis technique used and level of lymph node dissection. Conclusions: LDG is technically feasible and safe in morbidly obese patients with a BMI of ${\geq}30kg/m^2$ and early gastric carcinoma. Except for a longer operating time, LDG might represent a reasonable treatment option in these patients.

Clinical Study on the Successful Pregnancy and Delivery after Transfer of Human Blastocysts Cryopreserved by Vitrification (인간 포배란의 유리화동결 융해 후 임신 및 분만에 관한 연구)

  • Choi, Dong-Hee;Chung, Hyung-Min;Chung, Mi-Kyung;Lee, Sook-Hwan;Nam, Yoon-Seung;Park, Chan;Kwak, In-Pyung;Yoon, Tae-Ki
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.4
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    • pp.367-372
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    • 2000
  • Objective: This study was performed to evaluate whether vitrification method could be used for the cryopreservation of human blastocysts derived from IVF program. Methods: Surplus embryos were obtained from consented IVF patients. Controlled ovarian hyperstimulation was done with midluteal GnRH agonist, gonadotropin and hCG. After oocyte retrieval and insemination, fresh embryo transfer was done at $4{\sim}8$ cell stage. The surplus embryos after ET were cultured in blastocyst medium up to 6 days after oocyte retrieval. Obtained blastocysts were cryopreserved with our vitrification method. Blastocysts were exposed to 1.5 Methylene glycol (EG) in phosphate buffered saline (PBS) for 2.5 minutes, followed by 5.5 M EG plus 1 M sucrose for 20 seconds. Then 1 to 3 blastocysts were mounted on electron microscope (EM) grid and the grid was plunged into liquid nitrogen for storage. For thawing, blastocyst-containing EM grids were sequentially transferred in 1.0 M, 0.5 M, 0.25 M, 0.125 M and 0 M sucrose solution at the intervals of2.5 minutes. And blastocysts were cultured for about 6 hours and only re-expanded blastocysts were transferred to uterus of the patients on 4 to 5 days after ovulation in natural cycle or on 18 to 19 day of artificial cycle. Results: From Oct. 1998 to Jul. 1999, 34 patients were agreed to participate in this study. The mean age and duration of infertility of the patients were 31.6 years and 4.1 years, respectively. Among 34 cycles. replacements could be done in 20 cycles (58.8%). A total 93 blastocysts were thawed and 48 (51.6%) of them survived. Thirty-eight blastocysts, mean 1.9 embryos per patient, were transferred, resulting in 5 clinical pregnancies which consisted of 1 triplet, 2 sets of twins and 2 singleton pregnancies. The pregnancy rate per transfer was 25% and implantation rate was 23.6%. Five patients delivered 7 healthy babies including 2 sets of twins at term. Conclusion: Successful pregnancies and deliveries were established after transfer of vitrified human blastocysts. Vitrification using ethylene glycol as cryoprotectant and electron microscope grid is a rapid and simple method that can be effectively applied for the cryopreservation of human blastocysts.

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The effect of LH Receptor in the Pregnancy of Poor Responders (황체호르몬 수용체의 발현이 저반응 환자군의 임신에 미치는 영향)

  • Lee, Jung-Bok;Do, Byung-Rok;Kim, Eun-Soo;Kim, Myung-Hee;Chun, Eun-Kyung;Jeong, Hyeon-Jeong;Roh, Sung-Il;Kim, Moon-Kyoo;Yoon, Hyun-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.2
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    • pp.111-117
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    • 2004
  • Objectives: To investigate the effect of LH receptor in folliculogenesis, we confirm the expression level of LH receptor (LH-R) mRNA in human granulosa cells (GCs) and its expression levels were analyzed by comparison to embryo developmental rate and pregnancy rate. Materials and Methods: GCs were obtained at the time of oocyte retrieval from the patients undergoing IVF-ET program. The patients were divided into two groups: Group I (n=20) is poor responder (retrieved oocyte(s)$\leq$3ea), Group II (n=80) is normal responder (retrieved oocytes>3ea). After the extraction of total RNA, semiquantitative RT-PCR was performed and the expression level of LH-R mRNA was normalized by $\beta$-actin. Statistical analysis was performed by using $X^2$ test, Student's t-test and Pearson correlation. Results: In Group II, the relative values of LH-R mRNA (0.680 vs. 0.463, p<0.005) and pregnancy rate (54.7% vs. 23.1%, p<0.05) were significantly higher than in Group I. Number of retrieved oocyte(s) was gradually increased when the expression of LH-R mRNA was increased (p<0.05). But the quality of retrieved oocyte and transferred embryo were not related with the expression of LH-R mRNA. When the pregnancy rate was compared with FSH only group and FSH combined with hMG group in the ovarian stimulation protocol, FSH combined with hMG group was significantly higher than FSH only group in Group I (37.5% vs. 0%), and the expression of LH-R mRNA was significantly higher in hMG combined group than FSH only group (p<0.05). Conclusion: Expression level of LH-R mRNA has important role in ovarian function related with the response to gonadotrophin in human folliculogenesis. Furthermore these data might provide the evidence that additional use of hMG is helpful to poor responders.

Vitamin D Inhibits Expression and Activity of Matrix Metalloproteinase in Human Lung Fibroblasts (HFL-1) Cells

  • Kim, Seo Hwa;Baek, Moon Seong;Yoon, Dong Sik;Park, Jong Seol;Yoon, Byoung Wook;Oh, Byoung Su;Park, Jinkyeong;Kim, Hui Jung
    • Tuberculosis and Respiratory Diseases
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    • v.77 no.2
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    • pp.73-80
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    • 2014
  • Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.

Validation of the LC-MS/MS Method for Ginsenoside Rb1 Analysis in Human Plasma (LC-MS/MS를 이용한 인체 혈장에서 Ginsenoside Rb1의 분석법 검증)

  • Han, Song-Hee;Kim, Yunjeong;Jeon, Ji-Young;Hwang, Minho;Im, Yong-Jin;Lee, Sun Young;Chae, Soo-Wan;Kim, Min-Gul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.12
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    • pp.1753-1757
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    • 2012
  • A new liquid chromatographic tandem mass spectrometric (LC-MS/MS) assay for the quantification of ginsenoside Rb1 in human plasma was developed and validated. The separation was performed on a Agilent C18 column ($4.6mm{\times}150mm$, particle size 5 ${\mu}m$) with a gradient elution of 0.1% formic acid in water and 0.1% formic acid in methanol and a flow rate of 0.9 mL/min. The analyte was determined using electrospray positive ionization mass spectrometry in the multiple reaction monitoring (MRM) mode (m/z 1131.714${\rightarrow}$365.303). Human plasma samples were extracted with acetone : water (50:50) by the liquid-liquid extraction method. The method was linear over the dynamic range of 10~500 ng/mL with a correlation coefficient of r=0.9995. The intra-and inter-day precision over the concentration range of ginsenoside Rb1 was lower than 5.8% (correlation of variance, CV), and the accuracy was between 96.0~104.6%. This LC-MS/MS assay of ginsenoside Rb1 in human plasma is applicable for quantification in a pharmacokinetic study.

Predictors of Good Outcomes in Patients with Failed Endovascular Thrombectomy

  • Hyungjong Park;Byung Moon Kim;Jang-Hyun Baek;Jun-Hwee Kim;Ji Hoe Heo;Dong Joon Kim;Hyo Suk Nam;Young Dae Kim
    • Korean Journal of Radiology
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    • v.21 no.5
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    • pp.582-587
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    • 2020
  • Objective: Endovascular thrombectomy (EVT) fails in approximately 20% of anterior circulation large vessel occlusion (AC-LVO). Nonetheless, the factors that affect clinical outcomes of non-recanalized AC-LVO despite EVT are less studied. The purpose of this study was to identify the factors affecting clinical outcomes in non-recanalized AC-LVO patients despite EVT. Materials and Methods: This was a retrospective analysis of clinical and imaging data from 136 consecutive patients who demonstrated recanalization failure (modified thrombolysis in cerebral ischemia [mTICI], 0-2a) despite EVT for AC-LVO. Data were collected in prospectively maintained registries at 16 stroke centers. Collateral status was categorized into good or poor based on the CT angiogram, and the mTICI was categorized as 0-1 or 2a on the final angiogram. Patients with good (modified Rankin Scale [mRS], 0-2) and poor outcomes (mRS, 3-6) were compared in multivariate analysis to evaluate the factors associated with a good outcome. Results: Thirty-five patients (25.7%) had good outcomes. The good outcome group was younger (odds ratio [OR], 0.962; 95% confidence interval [CI], 0.932-0.992; p = 0.015), had a lower incidence of hypertension (OR, 0.380; 95% CI, 0.173-0.839; p = 0.017) and distal internal carotid artery involvement (OR, 0.149; 95% CI, 0.043-0.520; p = 0.003), lower initial National Institute of Health Stroke Scale (NIHSS) (OR, 0.789; 95% CI, 0.713-0.873; p < 0.001) and good collateral status (OR, 13.818; 95% CI, 3.971-48.090; p < 0.001). In multivariate analysis, the initial NIHSS (OR, 0.760; 95% CI, 0.638-0.905; p = 0.002), good collateral status (OR, 14.130; 95% CI, 2.264-88.212; p = 0.005) and mTICI 2a recanalization (OR, 5.636; 95% CI, 1.216-26.119; p = 0.027) remained as independent factors with good outcome in non-recanalized patients. Conclusion: Baseline NIHSS score, good collateral status, and mTICI 2a recanalization remained independently associated with clinical outcome in non-recanalized patients. mTICI 2a recanalization would benefit patients with good collaterals in non-recanalized AC-LVO patients despite EVT.

Improvement of Pregnancy Rates by Coculture of Human Embryos with Cumulus Cells in Glucose and Phosphate Free M-TALP Media (Glucose와 Phosphate가 제거된 M-TALP 배지에서의 난구세포 공배양에 의한 임신율 향상에 관한 연구)

  • Chung, B.S.;Chang, W.H.;Lee, M.H.;Kim, J.Y.;Bang, J.H.;Kim, K.H.;Suh, T.K.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.75-81
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    • 1999
  • The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

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Sentinel Lymph Node Identification Using $^{99m}Tc$-Neomannosyl Human Serum Albumin in Esophageal Cancer; Comparison with $^{99m}Tc$-Phytate (식도암 환자에서 $^{99m}Tc$-Neomannosyl Human Serum Albumin과 $^{99m}Tc$-Phytate를 이용한 감시림프절 탐색의 비교)

  • Kim, Hyun-Koo;Kang, Du-Young;Kim, Seung-Eun;Park, Jong-Jae;Jeong, Jae-Min;Mok, Young-Jae;Choi, Young-Ho
    • Korean Journal of Bronchoesophagology
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    • v.16 no.2
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    • pp.109-114
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    • 2010
  • Background: The aim of this study was to compare a novel mannose receptor-binding agent(Technetium-99m human serum albumin, $^{99m}Tc$-MSA) with $^{99m}Tc$-phytate for sentinel node mapping in patients with esophageal cancer. Material and Method: Twenty patients with clinical stage T1N0m0 or T2N0M0 esophageal cancer that were candidates for esophagectomy were enrolled. Endoscopic injection of $^{99m}Tc$-MSA or $^{99m}Tc$-phytate was administered at the peri-tumor region before surgery in 10 patients. The radioactive lymph nodes were identified with a handheld gamma probe after lymph node dissection. Results: The patient's age and the sex ratio of both groups were similar. The clinical stage, tumor location, and operative technique did not differ. The total number of dissected lymph nodes did not differ ($28.5{\pm}9.12$ in MSA group, $32.2{\pm}11.24$ in phytate group, p=0.430). The sentinel node was identified in all cases in both groups. The number of sentinel nodes per patient was $2.7{\pm}1.57$ in the MSA group, which was significantly greater than the $1.7{\pm}0.88$ in the phytate group (p=0.036). Five out of 20 patients whose sentinel nodes could be identified had metastases; however, neither group had any false-negative results for sentinel node identification. Conclusion: Sentinel nodes were detected more frequently with MSA than with phytate.

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