Cryo letters最新文献_第2页

A novel vitrified cryopreservation approach with stem cell-laden hydrogel microcapsules. 使用干细胞水凝胶微囊的新型玻璃化冷冻保存方法。

IF 1 4区生物学

Cryo letters Pub Date : 2024-03-01

T Song, B Liu

引用次数: 0

Optimizing semen cryopreservation in Calomys laucha: a step forward in rodent reproductive research. 优化 Calomys laucha 的精液冷冻保存：啮齿动物生殖研究的一大进步。

IF 1 4区生物学

Cryo letters Pub Date : 2024-03-01

C D Corcini, J Vilela, N Gatti, B Mion, N Castro, R T Franca, A S Varela Junior

{"title":"Optimizing semen cryopreservation in Calomys laucha: a step forward in rodent reproductive research.","authors":"C D Corcini, J Vilela, N Gatti, B Mion, N Castro, R T Franca, A S Varela Junior","doi":"","DOIUrl":"","url":null,"abstract":"Background: Examining semen cryopreservation in Calomys laucha offers valuable insights for reproductive research and species conservation.Objective: To determine the most effective sugar for the cryopreservation of C. laucha semen.Materials and methods: Using 36 epididymides from C. laucha, semen samples were diluted in a 3% skimmed milk medium supplemented with one of four sugars (glucose, fructose, lactose, or sucrose) at a concentration of 0.3 M. These mixtures underwent a conditioning phase at 37 degree C for 10 min, cooled to -80 degree C for another 10 min, and were subsequently stored in liquid nitrogen.Results: Upon thawing, samples treated with lactose and glucose solutions show superior sperm motility, achieving 8.2% and 10.0% respectively, in contrast to the fructose (2.0%) and sucrose (4.1%) mixtures. Furthermore, samples preserved in glucose registered the highest sperm penetration rates, reaching 44.9%.Conclusion: Our findings suggest that a cryopreservation medium containing 0.3 M glucose can contribute to the safeguarding C. laucha rodent semen. https://doi.org/10.54680/fr24210110612.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 2","pages":"134-138"},"PeriodicalIF":1.0,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140335074","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

RNA-Seq analysis reveals the mechanism in response to cold stress of peach cv 'Dingjiaba'. RNA-Seq 分析揭示了桃品种 "丁家坝 "对冷胁迫的响应机制。

IF 1 4区生物学

Cryo letters Pub Date : 2024-03-01

X Li, H Wang, X Wu

{"title":"RNA-Seq analysis reveals the mechanism in response to cold stress of peach cv 'Dingjiaba'.","authors":"X Li, H Wang, X Wu","doi":"","DOIUrl":"","url":null,"abstract":"Background: 'Dingjiaba' is an important Prunus persica cultivar (cv) mainly grown in the Hexi corridor in northwest China, which has an inherited strong cold tolerance.Objective: To compare the transcriptome and physiology data of leaves of cvs 'Dingjiaba' (D) and 'Kanoiwa' (K) following cold treatment at different time periods, in order to gain new insights into the mechanisms of cold adaptation in 'Dingjiaba'.Materials and methods: We analyzed the transcriptomic and physiological data of leaves of D and K cvs exposed to 0 h (D0/K0), 2 h (D2/K2), 6 h (D6/K6) and 12 h (D12/K12) cold stress.Results: Low temperature stress caused membrane damage and led to increased rate of electrolyte leakage and increased MDA content. Cold stress induced the accumulation of soluble sugars, soluble proteins and proline in leaves of both cvs, with a lower increase in K compared to D. Transcriptome analysis identified 4,631, 5,069, 5,662 and 3,886 differentially expressed genes (DEGs) between D0 and K0, D2 and K2, D6 and K6 and D12 and K12, respectively. The differentially expressed genes significantly enriched in metabolic pathways and biosynthesis of secondary metabolites. We further validated the reliability of sequencing data of the RNA-Seq with Real-Time Quantitative PCR, which suggested that the expression trend of the RNA-Seq were same as RT-PCR.Conclusions: These results provide novel insights into a series of molecular mechanisms underlying physiological metabolism and defense. https://doi.org/10.54680/fr24210110312.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 2","pages":"88-99"},"PeriodicalIF":1.0,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140335075","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Viscoelastic properties of decellularized and freeze-dried human dermis between i909c and 40cC. i909c 和 40cC 之间脱细胞和冻干人体真皮的粘弹性能。

IF 1 4区生物学

Cryo letters Pub Date : 2024-01-01

Q P Jia, S L Huang, Y T Tang, W Q Sun

{"title":"Viscoelastic properties of decellularized and freeze-dried human dermis between i909c and 40cC.","authors":"Q P Jia, S L Huang, Y T Tang, W Q Sun","doi":"","DOIUrl":"","url":null,"abstract":"Background: Human donor skin is processed to make the acellular dermis matrix (ADM) for tissue repair and regeneration. There is no data on the viscoelastic properties of ADM at room and subzero temperatures.Objective: The work evaluated the temperature dependence of viscoelastic properties of freeze-dried ADM.Materials and methods: Donor skin was de-epidermized, de-cellularized and freeze-dried with trehalose as the lyo-protectant. Glass transition of freeze-dried ADM was measured by differential scanning calorimeter (DSC), and viscoelastic properties were examined by dynamic mechanical analyzer (DMA).Results: At the low moisture range (1.4 +/- 0.5%), the glass transition temperature (Tg) of freeze-dried ADM was 90 degree C to 100 degree C. As the moisture content increased, the Tg decreased steadily. At the high moisture range (10.8 +/- 2.9%), the Tg was 40 degree C to 60 degree C. There were large donor-to-donor variations in viscoelastic properties of freeze-dried ADM as demonstrated by the changes in storage modulus (G'), loss modulus (G\") and damping factor tan delta (G\"/G'). However, the trends of the temperature dependence for G', G\" and tan delta were similar among all 8 donors. For each donor, changes in G' and G\" were relatively small between -90 degree C and 40 degree C, and G' was at least one order of magnitude greater than G\". Two viscoelastic relaxations were observed in freeze-dried ADM, one at -20 degree C and the other at -60 degree C respectively.Conclusion: Freeze-dried ADM was protected in the glassy carbohydrate matrix. DMA observed two viscoelastic relaxations (i.e., alpha process at -20 degree C and beta process at -60 degree C). Overall changes in G' and G'' of freeze-dried ADM were relatively small within one order of magnitude between -90 degree C and 40 degree C. https://doi.org/10.54680/fr24110110612.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 1","pages":"55-59"},"PeriodicalIF":1.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140305153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Supplementation of extender with chamuangone-enriched Garcinia cowa leaf extract improves quality parameters and longevity of cold-stored cat semen. 在扩增剂中添加富含辣椒素的加西牛肝菌叶提取物，可提高冷藏猫精液的质量指标和寿命。

IF 1 4区生物学

Cryo letters Pub Date : 2024-01-01

S Panyaboriban, N Kupthammasan, K Madsri, N Mukem, S Tarngwiriyakul, P Khirilak, P Panichayupakaranant, M Wittayarat

{"title":"Supplementation of extender with chamuangone-enriched Garcinia cowa leaf extract improves quality parameters and longevity of cold-stored cat semen.","authors":"S Panyaboriban, N Kupthammasan, K Madsri, N Mukem, S Tarngwiriyakul, P Khirilak, P Panichayupakaranant, M Wittayarat","doi":"","DOIUrl":"","url":null,"abstract":"Background: Semen preservation by cooling is less expensive, simpler and results in less sperm damage than freezing does. However, spermatozoa can only be preserved for a short period due to the excessive formation of reactive oxygen species (ROS). Although several antioxidants can protect sperms from ROS damage during storage at low temperatures, the use of natural antioxidants derived from plants would be a better alternative.Objective: To assess the effects of chamuangone, which can reduce oxidation reactions in cells, on cat semen quality after preservation at 4 degree C for 15 days.Materials and methods: Epididymal sperm samples were collected before being diluted with tris-citric-fructose-egg yolk (TCFE) extender containing different concentrations of chamuangone (0, 50, 100, 150 and 200 ug/mL) and preserved at 4 degree C. Semen samples were evaluated before chilling and then every 3 days after chilling for up to 15 days. Each sample was assessed for sperm motility, viability, DNA integrity, plasma membrane integrity and percentage of spermatozoa with intact acrosomes.Results: A significantly higher sperm motility was observed in the group supplemented with 100 ug/mL chamuangone compared to the control after 6 days of storage. However, the chamuangone concentration at 200 ug/mL did not significantly increase the sperm motility when compared to the control for the entire storage period.Conclusion: 100 µg/mL chamuangone can improve sperm characteristics during 15 days of preservation at 4 degree C, keeping sperm alive (49.3 ± 5.2%) and moving (7.1 ± 2.4%). These results can be used for the development of breeding programs using technologically advanced reproductive procedures in domestic and wild cats. https://doi.org/10.54680/fr24110110212.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 1","pages":"41-48"},"PeriodicalIF":1.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140305150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Dog sperm cryopreservation using cryovials and different dilution steps. 使用低温瓶和不同稀释步骤进行狗精子冷冻保存。

IF 1 4区生物学

Cryo letters Pub Date : 2024-01-01

S Ibrahim, N A Hamad Talha, J Cho, Y Jeon, I J Yu

{"title":"Dog sperm cryopreservation using cryovials and different dilution steps.","authors":"S Ibrahim, N A Hamad Talha, J Cho, Y Jeon, I J Yu","doi":"","DOIUrl":"","url":null,"abstract":"Background: The conventional sperm freezing method for dog sperm is with straws and includes two-step dilution and a long equilibration time.Objective: To develop a more efficient freezing method using cryovials.Materials and methods: Three freezing protocols using cryovials (0.5 mL) were conducted with dog spermatozoa at 1 x 108 sperm/mL: Group 1 spermatozoa were cooled in cryovials and extender 1 (E1) and extender 2 (E1 +1 M glycerol) at 4 degree C for 50 min and then frozen over LN2 for 20 min; Group 2 sperm was cooled and frozen in cryovials with a mixture of E1 and E2 (1:1) in a deep freezer (-80 degree C) for 30 min; Group 3 sperm in cryovials and E1 were cooled at 4 degree C for 20 min, cooled for an additional 20 min after addition of E2 (E1:E2, 1:1), and then frozen using LN2/ vapour for 20 min. The control (Group 4) consisted of spermatozoa in straws being frozen using the conventional freezing method using two-step dilution. All groups were plunged and stored in LN2 after freezing and their functional performance and gene expression determined.Results: Progressive motility and acrosome integrity were highest (P < 0.05) in Groups 2, 3 and 4 (only acrosome integrity). Viability in Group 3 was significantly better that in the other Groups, and the reactive oxygen species (ROS) level and phosphatidylserine (PS) translocation index were significantly lower in Group 2 than the other Groups. The expression of sperm mitochondria-associated cysteine-rich protein (SMCP) and anti-apoptotic B-cell lymphoma 2 (BCL2) genes was highest (P < 0.05) in Group 2 and the expression of pro-apoptotic Bcl2-associated X protein (BAX) was lowest (P < 0.05) in Group 4.Conclusion: The sperm frozen using cryovials, one step dilution and the deep freezer (Group 2) proved to be a simple and suitable cryopreservation method for dog sperm. https://doi.org/10.54680/fr24110110312.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 1","pages":"16-27"},"PeriodicalIF":1.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140305186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effects of stearic acid on the embryo cryopreservation in mouse. 硬脂酸对小鼠胚胎冷冻的影响

IF 1 4区生物学

Cryo letters Pub Date : 2024-01-01

T N Igonina, T A Rakhmanova, A N Omelchenko, K A Okotrub, E Yu Brusentsev, I N Rozhkova, S Ya Amstislavsky

{"title":"Effects of stearic acid on the embryo cryopreservation in mouse.","authors":"T N Igonina, T A Rakhmanova, A N Omelchenko, K A Okotrub, E Yu Brusentsev, I N Rozhkova, S Ya Amstislavsky","doi":"","DOIUrl":"","url":null,"abstract":"Background: Intracellular lipids are sensitive to freezing. Lipidome modification is an important tool for studying the role of intracellular lipids in cryotolerance of mammalian oocytes and preimplantation embryos.Objective: To study the effects of in vitro exposure of murine embryos to saturated stearic acid (SA) on the lipid content, embryo development and cryotolerance.Materials and methods: In vivo derived mouse embryos were cultured with 100 uM SA for 48 h up to the morula/blastocyst stage. Some of the SA-treated embryos were chosen for the evaluation of their development competence and the change in the lipidome, and other embryos were either slowly frozen or rapidly vitrified.Results: Nile red staining combined with confocal laser scanning microscopy revealed a decrease in the total amount of lipids in the SA-treated embryos. Raman measurements showed that the lipid unsaturation was lower in embryos after in vitro SA culture. The addition of SA did not affect the embryo development before cryopreservation, but negatively affected the results of slow freezing cryopreservation and vitrification.Conclusion: In vitro SA exposure lowered the total amount of intracellular lipids and unsaturation in mouse embryos. The changes were accompanied with a significantly lower efficacy of embryo cryopreservation. https://doi.org/10.54680/fr24110110512.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 1","pages":"28-35"},"PeriodicalIF":1.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140305187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The comet assay as a method for assessing dna damage in cryopreserved samples. 彗星试验作为一种评估低温保存样本中 dna 损伤的方法。

IF 1 4区生物学

Cryo letters Pub Date : 2024-01-01

B P Plitta-Michalak, A Ramos, D Stepien, M Trusiak, M Michalak

{"title":"The comet assay as a method for assessing dna damage in cryopreserved samples.","authors":"B P Plitta-Michalak, A Ramos, D Stepien, M Trusiak, M Michalak","doi":"","DOIUrl":"","url":null,"abstract":"The preservation of the nuclear genome's integrity is paramount for the viability and overall health of cells, tissues, and organisms. DNA, being susceptible to damage under physiological conditions and vulnerable to both endogenous and environmental factors, faces constant threats. To assess DNA damage and repair within individual eukaryotic cells, the comet assay presents itself as a versatile, gel electrophoresis-based, relatively simple, and highly sensitive method. Originally designed to monitor DNA damage and repair within populations of mammalian cells, the comet assay has now found applications across diverse domains, including yeast, protozoa, plants, and invertebrates. This technique has proven invaluable in cryopreservation studies, serving as a valuable adjunct for determining suitable cryopreservation protocols. These protocols encompass choices related to cryoprotectants, sample preparation, as well as storage conditions in terms of time and temperature. In the realm of animal cryopreservation research, the comet assay stands as a gold-standard method for assessing DNA integrity. Nevertheless, when applied in plant-oriented investigations, additional efforts are essential due to the distinct nature of plant cells and associated technical challenges. This review elucidates the fundamental principles underlying the comet assay, discusses its current iterations, and delineates its applications in the cryopreservation of both animal and plant specimens. Moreover, we delve into the primary challenges confronting the comet assay's utility as a monitoring tool in the context of plant sample cryopreservation. https://doi.org/10.54680/fr24110110112.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 1","pages":"1-15"},"PeriodicalIF":1.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140305151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The effect of different concentrations of laminarin on the quality of cryopreserved ram semen. 不同浓度的层粘蛋白对冷冻保存的公羊精液质量的影响。

IF 1 4区生物学

Cryo letters Pub Date : 2024-01-01

N Zangishhi, H Hajarian, H Karamishabankareh, L Soltani

{"title":"The effect of different concentrations of laminarin on the quality of cryopreserved ram semen.","authors":"N Zangishhi, H Hajarian, H Karamishabankareh, L Soltani","doi":"","DOIUrl":"","url":null,"abstract":"Background: Increasingly, sheep breeders are using artificial insemination to produce lambs, so finding methods that preserve ram sperm can be useful.Objective: To determine the protective effects of different concentrations of laminarin on ram sperm motility, viability, abnormalities, membrane, and DNA integrity, superoxide dismutase enzyme (SOD) activity, and malondialdehyde (MDA) production after freeze-thawing.Materials and methods: The ejaculates of four rams were collected and stored at 35 degree C. Semen samples were diluted with a tris-base extender containing 100, 200, 400, and 800 ug/mL of laminarin and a control extender containing no laminarin, then frozen in liquid nitrogen after 4 h in the refrigerator.Results: In the treatment of frozen-thawed spermatozoa with 800 ug/mL laminarin, motility, viability, membrane integrity, and DNA integrity were significantly higher than in the control. In spermatozoa that were exposed to 800 ug/mL laminarin after thawing, MDA production was significantly lower than in the control group. The percentage of abnormal spermatozoa in 800 ug/mL laminarin was significantly lower than that in the control.Conclusion: The addition of 800 ug/mL laminarin to the freezing extender increases motility, viability, SOD activity, and plasma membrane integrity, while reducing abnormality and MDA production in freeze-thawed ram semen. https://doi.org/10.54680/fr24110110812.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 1","pages":"60-68"},"PeriodicalIF":1.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140305152","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Reduced sperm number of murrah (Bubalus bubalis) bull semen in french mini-straw affects kinetic and functional competence after cryopreservation. 法式小吸管中布拉氏公牛精液的精子数量减少会影响冷冻保存后的动力学和功能能力。

IF 1 4区生物学

Cryo letters Pub Date : 2024-01-01

H P Yadav, T K Mohanty, R K Dewry, S A Lone, S Nath, M Bhakat, R K Baithalu, S Tiwari, D K Swain, P Kumar, A K Mohanty, T K Datta

{"title":"Reduced sperm number of murrah (Bubalus bubalis) bull semen in french mini-straw affects kinetic and functional competence after cryopreservation.","authors":"H P Yadav, T K Mohanty, R K Dewry, S A Lone, S Nath, M Bhakat, R K Baithalu, S Tiwari, D K Swain, P Kumar, A K Mohanty, T K Datta","doi":"","DOIUrl":"","url":null,"abstract":"Background: Extensive dilution of cattle semen with tris-based extender compromises certain sperm kinetic and functional traits following cryopreservation.Objective: To study sperm functions of buffalo bulls under high dilution rates.Materials and methods: Twenty-four ejaculates were harvested twice a week from four buffalo bulls, and diluted to sperm concentrations of 80, 60, 40 and 20 million/mL. Diluted samples were filled in straws, equilibrated at refrigeration temperature for 4 h, and frozen in liquid nitrogen. Frozen sperm samples were thawed for evaluation of kinetic and functional attributes.Results: Compared to 20 million/mL (million/mL) sperm sample, the total motility, progressive motility and rapid motility were reduced (P < 0.05) in 5 million/mL sample. The proportion of live sperm were significantly (P < 0.05) higher in 10, 15 and 20 million/mL samples than in 5 million/mL sample. The percentage of moribund sperm, dead sperm, and sperm with lipid per oxidation increased significantly (P < 0.05) in 5 million/mL sample.Conclusion: The reduction of sperm concentrations to < 10 million/mL affects post-thaw Buffalo sperm kinetic and functional attributes.. https://doi.org/10.54680/fr24110110712.","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"45 1","pages":"36-40"},"PeriodicalIF":1.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140305188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0