Effects of platelet-rich plasma on chondrocyte apoptosis and NLRP3/IL-1β signaling pathway in rabbits with osteoarthritis

AIM To explore the effect of platelet-rich plasma (PRP) on rabbit osteoarthritis and its possible mechanism. METHODS The rabbits with knee osteoarthritis were prepared and then divided into model group, sodium hyaluronate (SH) group and PRP group, and another sham operation group was set up, with 6 rabbits in each group. The gross morphological changes of rabbit cartilage were observed. HE staining was used to evaluate the pathomorphological changes of the cartilage. TUNEL staining was used to detect the apoptosis of chondrocytes. The expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/interleukin-1β （IL-1β） signaling pathway-related molecules was observed by immunohistochemical staining, and the protein levels of caspase-3, Bcl-2 and Bax were determined by Western blot. Chondrocytes were isolated and processed according to grouping, and the NLRP3 and IL-1β levels of the cells were measured by ELISA. RESULTS Compared with sham operation group, Pelletier score, Mankin score, chondrocyte apoptotic rate, the positive protein expression rates of NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax in model group were increased significantly (P<0.05), while the protein expression of Bcl-2 was decreased significantly (P<0.05). Compared with model group, Pelletier score, Mankin score, the apoptotic rate of chondrocytes, the positive protein expression rates of NLRP3, ASC, caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax in SH group and PRP group were decreased significantly (P<0.05), while the protein expression of Bcl-2 was increased significantly (P<0.05). In PRP group, Pelletier score, Mankin score, the apoptotic rate of chondrocytes, the positive protein expression rates of NLRP3, ASC, caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax were lower than those in SH group, while the protein expression of Bcl-2 was higher than that in SH group (P<0.05). Compared with control group, the expression of NL?RP3 and IL-1β in MCC950 (NLRP3 ihibitor) group were significantly reduced (P<0.05), the expression of NLRP3 in eucalyptol (IL-1β inhibitor) group was not significantly changed (P>0.05), and the expression of IL-1β was significantly reduced (P<0.05). CONCLUSION Platelet-rich plasma promotes the repair of cartilage in osteoarthritis rabbits, which has better effect than SH. The mechanism may be related to the inhibition of NLRP3/IL-1β pathway and the reduction of chondrocyte apoptosis.     

Mechanism of elemene enhancing radiosensitivity of human glioma U251 cells

AIM To investigate the effect of elemene on the radiosensitivity of human glioma U251 cells and its mechanism. METHODS The U251 cells were used as a glioma model in vitro, and were exposed to different concentrations of elemene and different doses of radiation. The cell viability was measured by MTT assay, the apoptosis and cell cycle distribution were analyzed by flow cytometry, and the related protein levels were determined by Western blot. RESULTS Elemene inhibited the viability of U251 cells in vitro and enhanced the radiosensitivity of the cells. The cells in radiotherapy combined with elemene group had higher rates of early apoptosis, secondary necrosis and total cell death than those in radiation group. Elemene induced G₂/M phase arrest in the U251 cells. Elemene reduced the protein expression of cell division cycle protein 2 （Cdc2）, which resulted in the decrease in cyclin B1 expression induced by radiotherapy, thereby inhibiting the formation of cyclin B-Cdc2 complex. Elemene reduced Cdc2 activity by inhibiting the phosphorylation of Cdc2 protein at threonine 161, thereby inducing G₂/M phase arrest in the cells. It also mediated apoptosis by down-regulating survivin expression. CONCLUSION Elemene may increase the sensitivity of U251 cells to radiotherapy by down-regulating Cdc2 protein, decreasing cyclin B1 expression, inhibiting the formation of cylcin B-Cdc2 complex and down-regulating the expression of survivin.     

ROCK promotes high glucose-induced cardiomyocyte apoptosis by inhi-biting PI3K/Akt signaling pathway

AIMTo investigate whether Rho-associated coiled-coil kinase (ROCK) is involved in high glucose-induced apoptosis of primary cardiomyocytes by regulating PI3K/Akt signaling pathway. METHODSPrimary Wistar rat cardiomyocytes were cultured and identified by α-sarcomeric actin (α-SCA) immunohistochemistry. Cardiomyocytes were treated with 5.5, 33 and 40 mmol/L glucose for 48 h. The cell viability was measured by MTT assay, and the mRNA expression of ROCK1 and ROCK2 in the cardiomyocytes was detected by RT-qPCR. Flow cytometry was used to analyze the apoptosis of the cardiomyocytes. The protein levels of ROCK1, ROCK2, cleaved caspase-3, Bcl-2, PI3K, Akt and p-Akt were determined by Western blot. In order to confirm the regulatory effect of ROCKs on PI3K/Akt signaling pathway, the cells were divided into control group (5.5 mmol/L glucose), high glucose group (33 mmol/L glucose) and high glucose+Y27632 (ROCK inhibitor) group. Western blot was used to detect the protein levels of ROCK1, ROCK2, PI3K, Akt and p-Akt. RESULTSAfter 48 h of high glucose exposure, the values of relative cell viability in 33 and 40 mmol/L glucose groups were (79.71±2.43)% and (68.41±7.49)%, respectively, both of which were significantly decreased compared with normal control group (P<0.05). After 48 h of high glucose exposure, the relative mRNA levels of ROCK1 and ROCK2 in 33 and 40 mmol/L glucose groups were significantly increased compared with normal control group (P<0.05). Compared with normal control group, the apoptotic rate in 33 and 40 mmol/L glucose groups was increased significantly (P<0.05). Compared with normal control group, the protein expression of ROCK1, ROCK2 and cleaved caspase-3 in 33 and 40 mmol/L glucose groups was increased (P<0.05), while the protein expression of Bcl-2 was decreased (P<0.05). No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed, while the protein level of p-Akt in 33 and 40 mmol/L glucose groups was decreased compared with normal control group (P<0.05). Compared with high glucose group, the expression of ROCK1 and ROCK2 was decreased in high glucose+Y27632 group. No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed. Compared with normal control group, the protein level of p-Akt in high glucose group was decreased, and the protein level of p-Akt in high glucose+Y27632 group was increased significantly compared with high glucose group. CONCLUSION Under high glucose environment, ROCK may reduce the level of p-Akt by inhibiting the PI3K/Akt signaling pathway, thus promoting the apoptosis of cardiomyocytes.     

甜樱桃园春季管理技术要点

从果园建立、整形修剪、土肥水管理、花果管理、病虫害防治和预防晚霜危害等方面介绍了甜樱桃园的春季管理要点,为同时做好疫情防控和果园管理提供参考。     

葡萄新品种妮娜女皇的引种表现和优质栽培技术

2014年引入妮娜女皇葡萄一年生苗木在淮安市园艺技术服务站果树示范基地试栽,经过5年试验观察,表现优良,经赤霉素和氯吡苯脲处理后,平均穗重680g,平均单果重17g,可溶性固形物含量平均18%,最高可达21%,甘甜可口,苗木栽植第4年每666.7m^2产量1000~1250kg。适应性强,丰产性良好,可推广。总结了妮娜女皇葡萄的优质栽培技术。     

氯虫苯甲酰胺水分散粒剂对桃小食心虫的防治效果研究

本试验比较了不同浓度氯虫苯甲酰胺水分散粒剂对桃小食心虫的防治效果。结果表明:试验药剂35%氯虫苯甲酰胺水分散粒剂在水中分散性良好,700倍用药量(有效成分量为41.2 g/hm^2)防治桃小食心虫的速效性和持效性均良好,14 d后防效达到最大值,为99.1%。试验证明,氯虫苯甲酰胺水分散粒剂可用来防治果树桃小食心虫。     

槲树叶提取液对冷藏期间桃品质及抗氧化系统的影响

以生理成熟期‘新泰红’桃果实为试材,分别用0、2.5、5.0、7.5 g/L槲树叶提取液浸泡果实,以蒸馏水为对照,并于0℃冷藏,测定冷藏期间果实可溶性蛋白等品质指标以及酶活力与活性氧等物质,探讨槲树叶提取液对采后桃果实贮藏品质的影响。结果表明:槲树叶提取液处理能够有效延缓冷藏期间桃果实冷害指数升高以及硬度下降,使整个冷藏期间果实可溶性蛋白、可溶性糖、维生素C含量维持相对较高水平,并且能够有效延缓膜脂过氧化产物(丙二醛)以及活性氧物质(超氧阴离子自由基、过氧化氢和羟基自由基)积累;同时,槲树叶提取液处理还能够显著抑制果实多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)活性,从而延缓冷藏期间桃果实褐变程度。综上所述,槲树叶提取液处理能够缓解冷藏期间桃果实冷害症状,以5.0 g/L槲树叶提取液处理效果最为明显。     

甘肃成县核桃种质资源晚霜冻害调查

为了解核桃种质资源对晚霜冻害的抗性表现,筛选抗晚霜冻的核桃种质资源,为抗晚霜冻核桃品种的选育提供优异材料,于晚霜冻害发生后的第3~5 d分别对成县当地核桃品种(系)、引种国内核桃品种、引种国外核桃品种、黑核桃及山核桃属的长山核桃等30份不同核桃种质的抗晚霜冻情况进行了田间调查和鉴定。结果表明:成县当地核桃品种(系)中,‘陇南755’、Y1102、Y1201等表现为抗晚霜冻型,B0802-8、Y1001萌芽较迟,属于避晚霜冻型;引种国内品种中,‘辽核4号’和‘辽核7号’表现为抗晚霜冻型;引种国外品种中,日本核桃‘清香’属于抗晚霜冻型,美国核桃种质‘强特勒’‘土莱尔’‘美国黑核桃’等属于避晚霜冻型,‘维纳’‘美国819’‘美国长山核桃’等属于抗或高抗晚霜冻型。     

不同介质对真姬菇菌种保藏效果的对比

选择适宜的菌种保藏介质,是有效保障真姬菇菌种品质的重要一环,不仅关系到真姬菇栽培者的直接利益,也关系到种质资源的保存与延续。通过测定菌丝生长速率和菌丝脱氢酶活力,对22种真姬菇菌种保藏介质的保藏效果进行了对比分析。结果表明0.1%的PEG6000溶液对真姬菇菌种保藏6个月和9个月的整体保藏效果最好,其次是0.1%的PEG10000溶液与超纯水,再次是0.8%葡萄糖溶液和矿泉水。0.6%葡萄糖溶液仅适于真姬菇菌种9个月的保藏,纯净水仅适于真姬菇菌种6个月的保藏。其他介质的保藏效果略差。     

不同处理方法对不同澳洲坚果品种嫁接试验研究

为了提高澳洲坚果育苗的成活率,确保种苗供应。对桂热1号、Hinde(H2)、Pahala(788)、O.C、344等5个澳洲坚果品种枝条分别采用环剥、萘乙酸、环剥+萘乙酸处理后进行嫁接试验。结果表明,环剥+萘乙酸处理5个澳洲坚果品种的平均嫁接成活率较对照提高了42.9%,新梢较对照多3.9条,新梢长度较对照长18 cm;环剥处理平均嫁接成活率较对照提高了33.6%,新梢较对照多3.3条,新梢长度较对照长14.16 cm;萘乙酸处理平均嫁接成活率较对照提高了24.6%,新梢较对照多2.5条,新梢长度较对照长3.92 cm。以环剥+萘乙酸处理嫁接效果最好。