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1.
牛体外受精胚胎一步脱防冻剂冷冻方法的研究   总被引:7,自引:1,他引:7  
为了研究适合于牛体外受精胚胎的一步除防冻剂的冷冻方法,特进行两个试验。试验1分别用1.5M甘油+0.25M蔗糖、1.5M乙二醇和1.5M丙二醇作防冻剂冷冻体外受精后第7天,已发育至囊胚阶段的牛胚胎。使胚胎降温至-7℃后,植冰,然后以0.3℃/分的速率降温至-30℃,立即将胚胎投入液氮中冷冻保存。在37℃水中解冻胚胎后,使其直接在含15%胎犊血清(FCS)的磷酸缓冲液(PBS)中脱去防冻剂。经体外培养72h后,3组胚胎的孵化率分别为77.27%(102/132)、73.24%(104/142)、47.90%(57/119),第1、2组的孵化率极显著高于第3组(P<0.01),说明用1.5M丙二醇溶液冷冻的胚胎,在解冻后不宜直接在PBS中脱防冻剂。试验2比较用不同浓度(1.0M01.5M,2.0M)的乙二醇和不同投液氮温度(-25℃,-30℃,-35℃)冷冻胚胎的效果。结果表明乙二醇浓度和投液氮温度对胚胎孵化率均无显著影响(P>0.05),各组胚胎的孵化率变动于74.44%-85.48%之间。  相似文献   

2.
The heart rates (HR) and rhythms of 15 healthy donkeys of various ages, both sexes, and various breeds were analyzed throughout 24-hour Holter monitoring. The animals were evaluated at rest in their daily environment without the presence of investigators causing stress, using a three-channel digital Holter recorder. Analysis revealed a maximal HR range from 62.5 to 93.7 beats/min (mean, 72.50 ± 7.51 beats/min), whereas the minimal HR ranged from 29.7 to 42.2 beats/min (mean, 34.90 ± 4.22 beats/min). The daily mean HR was 47.55 ± 3.70 beats/min. The daytime mean HR was 49.39 ± 2.77 beats/min, whereas the night time rate was 46.22 ± 4.38 beats/min. Sinus bradycardia and sinus tachycardia were detected in 7 and 10 of 15 studied donkeys, respectively. Cardiac dysrhythmias due to high vagal tone such as sinoatrial heart block and second-degree atrioventricular heart block were occasionally recorded in 1 and 3 donkeys, respectively. A higher mean HR and fewer cardiac dysrhythmias were observed in donkeys than in horses and ponies at rest. These findings could be justified by differences in the autonomic nervous system tone.  相似文献   

3.
A rapid incremental treadmill exercise test was used to determine the repeatability of the following measurements in exercising horses: maximal oxygen consumption (VO2max), maximal heart rate (HRmax), velocity at a heart rate of 200 beats/min (V-200), oxygen consumption at a heart rate of 200 beats/min (VO2-200), oxygen consumption at HRmax (VO2-HRmax), work rate at a heart rate of 200 beats/min (W-200), work rate at HRmax (W-HRmax) and treadmill velocity at HRmax (V-HRmax). Six Standardbred geldings were exercised on three separate occasions on a treadmill set at an inclination of 6 degrees. The exercise protocol was that each horse was exercised for 2 mins at 3 m/sec, after which the treadmill speed was increased by 1 m/sec every 60 secs, until the horse could no longer maintain its speed. A minimum of 24 h was allowed between repeated tests. No significant differences were found between the three means of any of the eight cardiorespiratory variables with repeated measurement. Variables with high coefficients of variation (greater than 10 per cent) included V-HRmax, W-HRmax and VO2-HRmax. The V-200, W-200 and VO2-200 showed less variation. The VO2max showed good reproducibility, there being coefficients of variation ranging from 1.4 to 9.0 per cent. The individual horse values for VO2max ranged from 104 to 169 ml/kg bodyweight/min. Maximal heart rate was also highly reproducible and the coefficients of variation were less than or equal to 2.7 per cent in all horses. It is concluded that the measurement of VO2max has good reproducibility, but other estimates of maximal aerobic capacity are less precise.  相似文献   

4.
This study evaluated the effectiveness of glycopyrrolate (0.005 or 0.01 mg/kg body weight (BW)) in anesthetized dogs (n = 40) for reversal of bradycardia (< 65 beats/min). Following random intravenous (i.v.) treatment, heart rate was determined at 5 min and, if it was < or = 70 beats/min, the lower dose was repeated. A 2-way analysis of variance considered dose and animal size (< or = 10 kg, > 10 kg) effects (P < 0.05). Glycopyrrolate produced a significant increase in heart rate and infrequent tachycardia (< or = 150 beats/min), which was not dose-related. The size of the dog produced a significant effect on baseline heart rate (higher in small), rate following the first dose (lower in small), and requirement for retreatment (47% in small, 13% in large). In a separate group of anesthetized dogs (n = 20), the blood pressure effect of glycopyrrolate (0.01 mg/kg BW, i.v.) treatment of bradycardia (65-85 beats/min, weight-adjusted) was studied. A significant increase in systolic, diastolic, and mean blood pressure was produced. In conclusion, the effective dose of glycopyrrolate treatment is size-related and produces a beneficial effect on blood pressure.  相似文献   

5.
本文主要研究了①离子霉素(Ion)+6-二甲基氨基嘌呤(6-DMAP)、②Ion+放线菌酮(CHX)、③体积分数7%乙醇+6-DMAP、④体积分数7%乙醇+CHX四种不同化学激活方法对牛胞质内卵丘细胞全细胞注射法所获得重构胚的激活及前期发育的影响。Ion和7%乙醇的处理时间为5min,6-DMAP的处理时间为4h.CHX的处理时间为5h。结果表明,重构胚与颗粒细胞单层细胞共培养2d后,各组卵裂率分别为52.7%、51.5%、53.2%和54.0%.差异不显著(P〉0.05);培养8d后,①组激活的囊胚发育率(20.0%)极显著高于其他组(P〈0.01),其他各组之间差异不显著.分别为8.5%、10.2%和6.1%;培养10d后,①组激活的囊胚孵化率(10.7%)极显著高于其他组(P〈0.01).其他各组之间差异不显著,分别为2.3%、3.0%和1.8%。表明,4种激活方法均可以有效地激活重构胚,但Ion+6-DMAP法激活重构胚的囊胚率和囊胚孵化率均极显著高于其他3组,说明Ion+6-DMAP激活法有利于牛重构胚的发育.可以获得较理想的囊胚率.是胞质内全细胞注射法克隆的理想激活方法。  相似文献   

6.
ABSTRACT

1. Sex-specific variations in early embryonic development rates may pre-empt later variations in embryonic development through to pipping and hatching. Given that erythropoiesis (blood production) can be equated with early embryonic growth rate, it was hypothesised that blood pigment haemoglobin can act as a specific spectral fingerprint for changes in growth rate. Moreover, by measuring longitudinal, rather than lateral, spectral transmission through the egg, a more consistent spectrum with a higher signal-to-noise ratio could be captured.

2. Longitudinal visible transmission (T575/T598 ratio), which is sensitive to haemoglobin, was used to monitor sex-specific early embryonic development rate in white layer chicken eggs from d 0 to 8 of incubation. The sex of these eggs was subsequently confirmed two days after hatching.

3. Embryonic development was detectable from d 3 (72 h) of incubation, 36 h earlier than previously reported lateral spectral measurements, supporting the greater sensitivity of longitudinal measurements.

4. At d 3, the mean T575/T598 ratio for male embryos was significantly lower (P < 0.001) (i.e. higher absorbance of haemoglobin) than for female embryos, which was thought to be due to sex-differences in early embryogenesis. On the other hand, female embryos had a significantly lower (P < 0.05) mean T575/T598 ratio than male embryos at d 7 of incubation, presumably due to the combined effects of oestrogen synthesis receptors and enzymes on erythropoiesis in female embryos at this time.

5. In conclusion, the proposed methodology has the sensitivity to differentiate sex-specific embryonic development rates during early incubation and the potentiality to advance precision incubation management and poultry research.  相似文献   

7.
Primordial germ cells (PGCs) are embryonic precursors of germline cells with potential applications in genetic conservation, transgenic animal production and germline stem cell research. These lines of research would benefit from improved germline transmission of transplanted PGCs in chimeric chickens. We therefore evaluated the effects of pretransplant X-irradiation of recipient embryos on the efficacy of germline transmission of donor PGCs in chimeric chickens. Intact chicken eggs were exposed to X-ray doses of 3, 6 and 9 Gy (dose rate = 0.12 Gy/min) after 52 h of incubation. There was no significant difference in hatching rate between the 3-Gy-irradiated group and the nonirradiated control group (40.0 vs. 69.6%), but the hatching rate in the 6-Gy-irradiated group (28.6%) was significantly lower than in the control group (P<0.05). No embryos irradiated with 9 Gy of X-rays survived to hatching. X-irradiation significantly reduced the number of endogenous PGCs in the embryonic gonads at stage 27 in a dose-dependent manner compared with nonirradiated controls. The numbers of endogenous PGCs in the 3-, 6- and 9-Gy-irradiated groups were 21.0, 9.6 and 4.6% of the nonirradiated control numbers, respectively. Sets of 100 donor PGCs were subsequently transferred intravascularly into embryos irradiated with 3 Gy X-rays and nonirradiated control embryos. Genetic cross-test analysis revealed that the germline transmission rate in the 3-Gy-irradiated group was significantly higher than in the control group (27.5 vs. 5.6%; P<0.05). In conclusion, X-irradiation reduced the number of endogenous PGCs and increased the germline transmission of transferred PGCs in chimeric chickens.  相似文献   

8.
1. Hatching time, hatchability of fertile eggs and embryo mortality under standard egg storage (1 or 5 days at 12?±?1°C and 55% relative humidity) and incubation conditions (37·5?±?0·2°C and 50% relative humidity) were analysed in lines long-term selected for high (HG) and low (LG) relative weight gain between 11 and 28?d of age, respectively, and constant body weight at 49?d of age.

2. Egg storage duration did not have an effect on average hatching time. LG quail, characterised by a fast postnatal growth rate immediately after hatching, hatched earlier than HG quail with a low early growth rate (about 391 vs. 406?h after egg setting, respectively).

3. In contrast to hatching time, the hatchability of fertile eggs was influenced by line as well as egg storage duration. Extended storage decreased hatching success in both lines. However, LG eggs exhibited a higher hatchability than HG eggs (1?d storage: 96·0 vs. 82·5%; 5?d storage: 88·7 vs. 72·7%, respectively).

4. Lower hatchability resulted mostly from a higher frequency of embryo death during early (up to d 7) and late (d 14 and later) phases of incubation.

5. An inadequate nutrient supply to embryos in consequence of developmental delay seems to be a key factor decreasing viability of embryos during incubation.  相似文献   

9.
This study examined the effects of different vitrification medium compositions and exposure times (2, 4 and 6min) on the post-thaw development of buffalo embryos produced in vitro (IVP). The compositions were (1) 40% ethylene glycol (EG); (2) 25% glycerol (G)+25% EG, and (3) 25% EG+25% dimethylsulfoxide (DMSO). The base medium was 25mM Hepes-buffered TCM-199+10% steer serum +50microg/mL gentamycin. The IVP embryos were cryopreserved by a two-step vitrification method at 24 degrees C. After warming, the embryos were cultured in vitro for 72h. The vitrification of morulae and blastocysts in 25% EG+25% DMSO with an exposure time of 2 and 4min, respectively, resulted in a better hatching rate than other combinations. The hatching rate of morulae vitrified in 25% EG+25% G, 25% EG+25% DMSO, and blastocysts vitrified in 40% EG, 25% EG+25% DMSO were negatively correlated with exposure time. However, the hatching rate of blastocysts vitrified in 25% EG+25% G was positively correlated with exposure time. The study demonstrated that the post-thaw in vitro development of IVP buffalo embryos was affected by the vitrification medium composition and exposure time.  相似文献   

10.
The purpose of this study was to determine the heart rate (HR) and blood pressure (BP) effect of glycopyrrolate in anesthetized horses with low HR (< or = 30 beats/min). The horses were randomly treated with glycopyrrolate (2.5 micrograms/kg body weight (BW)) or saline, intravenously (i.v.) (n = 17). If HR failed to increase (by > 5 beats/min within 10 min), glycopyrrolate (same dose) was administered. Heart rate increased by > 5 beats/min in 3 out of 9 horses following the initial glycopyrrolate treatment. Overall changes in HR and mean BP were not significantly different, while systolic and diastolic BP increased significantly (P < 0.025 using a Bonferroni corrected paired t-test). On the 2nd treatment, 3 out of 7 horses given 2.5 micrograms/kg BW glycopyrrolate, and 4 out of 5 horses given 5.0 micrograms/kg BW (total dose) showed an increase in heart rate of > 5 beats/min, which was significant. A significant increase in BP was produced following treatment with 2.5 micrograms/kg BW, but not following 5.0 micrograms/kg BW. A final increase in HR, of > 5 beats/min, was associated with a significant rise in BP (P < 0.05 using an unpaired t-test). In conclusion, an increase in HR can occur with 2.5 to 5.0 micrograms of glycopyrrolate/kg BW, i.v., and results in improvement in BP in anesthetized horses.  相似文献   

11.
The present study was conducted to examine post-thaw in vitro developmental competence of buffalo embryos cryopreserved by cytoskeletal stabilization and vitrification. In vitro produced embryos were incubated with a medium containing cytochalasin-b (cyto-b) in a CO2 incubator for 40 min for microfilament stabilization and were cryopreserved by a two-step vitrification method at 24℃ in the presence of cyto-b. Initially, the embryos were exposed to 10% ethylene glycol (EG) and 10% dimethylsulfoxide (DMSO) in a base medium for 4 min. After the initial exposure, the embryos were transferred to a 7 µl drop of 25% EG and 25% DMSO in base medium and 0.3 M sucrose for 45 sec. After warming, the embryos were cultured in vitro for 72 h. The post-thaw in vitro developmental competence of the cyto-b-treated embryos did not differ significantly from those vitrified without cyto-b treatment. The hatching rates of morulae vitrified without cyto-b treatment was significantly lower than the non-vitrified control. However, the hatching rate of cyto-b-treated vitrified morulae did not differ significantly from the non-vitrified control. This study demonstrates that freezing of buffalo embryos by cytoskeletal stabilization and vitrification is a reliable method for long-term preservation.  相似文献   

12.
Vitrification could provide a promising tool for the cryopreservation of fish embryos. However, to achieve cryopreservation using vitrification, chilling sensitivity and cryoprotectants toxicity were determined using tench embryos at four developmental stages (11, 17, 23 and 29 h). Embryos treated with alcalase (2 ml/998 ml, 2 min at 22°C) were exposed to chilling with/without warming. Other embryos were exposed to methanol and glycerol at the concentration of 10% and 20% for periods of 20 min. At last, embryos were incubated at special incubator cages where hatching rates were counted. Regarding chilling sensitivity and exposure to chilling followed by warming, the hatching rates of embryos decreased significantly (p < 0.001) after exposure to 0°C at all developmental stages except the 29-h stage compared with the controls. The embryo stage most sensitive to chilling was 11-h stage. The 29-h stage exhibited the least sensitivity to low temperature while 17-h and 23-h stages were intermediate in their sensitivity to chilling. The toxicity of methanol increased significantly (p < 0.001) with developmental stage for 11, 17 and 23-h stages. The highest hatching rates of tench embryos were obtained with 29-h embryos using various concentrations of methanol. The hatching rates of tench embryos exposed to glycerol concentrations were approximately similar to those embryos exposed to methanol concentrations except for 11-h embryos that showed no hatching. Unfortunately, we could not obtain living embryos in any of the conditions examined after vitrification. In conclusion, it was quite difficult to vitrify the tench embryos during this study using various vitrifying solutions and the method reported by Chen & Tian (2005) and further studies are needed to achieve successful cryopreservation.  相似文献   

13.
1. Daily changes in embryonic heart rate (HR) of emu were determined non-invasively at 36 degrees C by acoustocardiography (ACG) during the last 30% of artificial incubation (predicted incubation time is 50 d). 2. The pattern of daily changes in mean HR of hatched embryos decreased from about 175 bpm to about 140 bpm towards the end of incubation. 3. The mean HR at 80% of incubation (ca. 170 bpm) was close to the value predicted from an allometric equation reported previously for precocial domesticated birds. 4. ACG could measure embryonic HR even during the external pipping period. 5. If the artificial external pipping procedure is timed correctly after internal pipping, it might aid the embryos in hatching. However, further investigation into this aspect is needed.  相似文献   

14.
A 20-min van journey increased plasma cortisol concentrations to 15-25 ng/ml in male goats, blood glucose concentrations were not affected, but respiratory rates and heart rates were increased, the latter by 40 beats per min. A 2-h van journey increased plasma cortisol to greater than 25 ng/ml and blood glucose to greater than 5 mmol/l. Respiratory rates were increased to greater than 40 breaths per min and heart rates by greater than 100 beats per min. Xylazine alone (0.01 mg/kg) suppressed resting plasma cortisol concentrations, increased blood glucose concentrations to 4.5 +/- 0.8 mmol/l and suppressed respiratory rates by 5-10 breaths per min and heart rates by 20 beats per min. Cortisol concentrations were suppressed by xylazine treatment if given before a 20-min van journey, and for approximately 60 min if given 20 min after the start of a 2-h journey. When combined with transport, xylazine caused an additive effect on glucose concentrations, but suppressed respiratory and heart rates. However, for the latter criteria the timing of suppression was different depending on the time of onset and duration of the stressor. Injection of 50 micrograms ovine corticotrophin releasing factor (CRF) caused an immediate elevation of cortisol concentrations (but not glucose) which lasted for at least 6 h compared with the return to baseline within 60 min after either length of journey. Xylazine pretreatment did not alter the cortisol response to CRF, suggesting that xylazine must act centrally above pituitary level when blocking the cortisol response to transport. It is proposed that under resting conditions the hypothalamus is under alpha 2-adrenergic suppression. Stimulation of cortisol secretion in response to a stressor can be inhibited by an alpha 2-adrenergic agonist.  相似文献   

15.
1. Using a flexible piezoelectric probe, we detected non-invasively the cardiogenic precordial movements of domestic fowl chicks and measured heart rate (HR) daily over the first week after hatching (day 0) for comparison with previously measured embryonic HR. 2. HR of the hatchlings was much more variable than embryonic HR. The mean HR at a given age also varied among the 5 chicks measured. 3. The overall mean HR was 280 +/- 20 (SD, n = 5) bpm on day 0. This value was similar to the prepipping HR and lower than that of externally pipped embryos. It increased to 342 +/- 39 bpm 3 days later.  相似文献   

16.
OBJECTIVE: To determine whether intravenous infusion of nitroglycerin would modify pulmonary arterial, capillary, or venous hypertension in strenuously exercising Thoroughbreds. ANIMALS: 5 healthy Thoroughbred horses. PROCEDURE: Right atrial, right ventricular, and pulmonary vascular pressures were measured. Each horse was used in a control treatment (not medicated) and a nitroglycerin infusion (20 microg/kg of body weight/min) at rest and during exercise on a treadmill. Sequence of treatments was randomized for each horse, and treatments were separated by a 7-day interval. Galloping at 14.2 m/s on a 5% uphill grade elicited maximal heart rate (mean +/- SEM, 212 +/- 2 beats/min) and could not be sustained for > 90 seconds. Nitroglycerin dosage was selected, because maximal pulmonary and systemic hemodynamic effects of i.v. nitroglycerin were elicited at 5 microg/kg/min and increasing the dosage to 20 microg/kg/min did not cause adverse effects. RESULTS: In the control treatment, exercise performed at maximal heart rate resulted in a significant increase in right atrial as well as pulmonary arterial, capillary, and wedge pressures. Nitroglycerin infusion in standing horses significantly decreased right atrial and pulmonary vascular pressures, whereas heart rate increased. Exercise in nitroglycerin-infused horses also resulted in a significant increase in right atrial as well as pulmonary arterial, capillary, and wedge pressures, and these values were not significantly different from data for the control treatment. All horses experienced exercise-induced pulmonary hemorrhage for both treatments. CONCLUSIONS AND CLINICAL RELEVANCE: I.v. administration of nitroglycerin does not modify exercise-induced pulmonary hypertension and is unlikely to affect the incidence or severity of exercise-induced pulmonary hemorrhage in Thoroughbreds.  相似文献   

17.
The well recognized cardiac effects of severe hyperkalemia include progressive rhythm and conduction disturbances such as bradycardia, spiked and narrow T waves, widening QRS complex, widening and flattening P wave, disappearance of the P wave, and cardiac arrest. Paradoxically, a heart rate greater than 200 beats/min may coexist with hyperkalemia in some cats. This report describes three cats with moderate to severe hyperkalemia and concurrent rapid heart rate. In each cat, the serum potassium (K(+)) concentration was > or =7.5 mEq/dl with a concurrent heart rate > 200 beats/min. In each cat, nine-lead electrocardiograms demonstrate an absence of P waves and a wide-complex tachycardia. Hyperkalemia should be considered in the differential diagnosis when a feline electrocardiogram demonstrates a wide-complex tachycardia without identifiable P waves.  相似文献   

18.
Knowledge of the competitive demands of different sports or activities is important for designing appropriate training programmes to ensure that animals reach a sufficient level of fitness to reduce the risk of overexertion and injury or illness and to achieve the best possible performance in relation to an individual's genetic potential. Whilst the physiological demands of many equestrian sports have been described, to the best of our knowledge the cardiovascular demands of polo have not. The aims of the present study were therefore to record heart rate during and after competitive polo games in a group of low-goal (non-elite) polo ponies in order to describe the absolute heart rates during play, the relationship of these heart rates to maximal heart rate and the characteristics of a typical chukka in terms of effort. Six low-goal polo ponies were studied during a total of 59 chukkas. Heart rate was monitored continuously before, during and after competition using a commercial heart rate monitor. Maximal heart rate was determined with field and treadmill incremental exercise tests and used to express work intensity in terms of time during play that each ponies heart rate was less than 75% HRmax, between 75 and 90% HRmax and greater than 90% HRmax. Mean maximum heart rate was not different during play or during field and treadmill exercise tests; 215+/-7 (mean +/- s.d.), 211+/-7 and 213+/-2 beats/min, respectively (P>0.05). Mean heart rate for all ponies over all chukkas was 166+/-6 beats/min with a mean chukka duration of 611+/-18 s. Of this time, 44+/-7% of the time was spent below 75% HRmax, 39+/-8% between 75 and 90% HRmax and 17+/-8% of time above 90% HRmax. When only one chukka had been played, there was a good correlation between mean heart rate during play and 3 min recovery heart rate (r = 0.63, P<0.001). Based on these observations, it is proposed that low-goal polo places moderate to high stress on the cardiovascular system.  相似文献   

19.
Seventy-two crossbred gilts were utilized to examine whether the oviduct rendered zona pellucidae resistant to protease digestion, whether the uterus reversed this resistance and whether such a uterine reversal was necessary for hatching. Oocytes were aspirated from follicles 22 to 28 h after onset of estrus (d 0); oviductal and uterine oocytes or embryos were collected on d 1 to 6. These oocytes and embryos were subjected to a solution containing .1% trypsin and .1% pronase (37 degrees C) for observation of zona pellucidae digestion. Zonae of oviductal oocytes were more (P less than .001) resistant to digestion than were follicular oocytes. Placement of follicular oocytes in oviducts for 30 min rendered zona pellucidae more (P less than .001) resistant to protease digestion than oocytes not exposed to oviductal secretions. Resistance of zona pellucidae to proteases, however, decreased (P less than .001) after entry into the uterus. Zonae of morulae retained in oviducts took longer (P less than .001) to digest than those recovered from the uterus. Blastocysts also were treated with wheat germ agglutinin (WGA; 50 micrograms/ml) for 40 min to determine whether artificial induction of zona resistance to enzymatic digestion affected the ability of embryos to hatch. Though WGA treatment delayed (P less than .001) enzymatic digestion of zona pellucidae, time from collection to hatching was not affected. This experiment indicated that the oviduct delayed enzymatic digestion of the zona pellucidae, whereas the uterus reversed this delay. The re-establishment of enzyme susceptibility after uterine entry, however, appeared to be unrelated to the subsequent ability of blastocysts to hatch.  相似文献   

20.
为研究CRISPR/Cas9腺病毒载体在鸡胚中进行基因敲入的可行性,将包装不同滴度增强型绿色荧光报道基因(Enhance green fluorescent protein,EGFP)的腺病毒载体和慢病毒载体显微注射到HH14时期鸡胚的外周血管中,对胚胎发育至3.5 d和9d鸡胚存活、各器官中EGFP荧光强度等指标进行...  相似文献   

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