响应面法优化超声波辅助提取薏苡仁低聚糖工艺的研究

采用响应面法优化超声波辅助提取薏苡仁低聚糖的工艺条件。在单因素试验基础上,选取液料比、超声波时间以及超声波功率3个因素结合Box-Behnken试验建立数学模型,分析考察3个因素对薏苡仁低聚糖响应值的影响程度,优化工艺参数。各因素对薏苡仁低聚糖提取率影响程度从大到小顺序依次为:超声波功率超声波时间液料比。响应面设计法优化出其最佳超声波提取条件为:超声波温度70℃,液料比33∶1(m L/g),超声波时间27 min,超声波功率450 W。在该条件下,薏苡仁低聚糖提取率为0.94%,与模型预测值0.98%接近。说明使用响应面法优化超声波辅助提取薏苡仁低聚糖的工艺条件是可行的。     

马铃薯AP2/ERF转录因子的克隆及植物表达载体构建

AP2/ERF基因家族转录因子普遍存在于植物中,参与植物体内的各种生物学过程,包括植物的生长发育、生物和非生物胁迫响应等。前期转录组测序的结果发现马铃薯‘加湘1号’一个AP2/ERF家族基因(PGSC0003DMG400012154)在接种晚疫病菌(Phytophthora infestans)24 h后被显著激活。从接种P.infestans 24 h的‘加湘1号’的总RNA中通过RT-PCR获得了该基因CDS序列为885 bp,BLAST分析显示该基因编码一个295个氨基酸残基的蛋白,并且含有1个AP2/ERF结构域,是AP2/ERF转录因子家族中的ERF亚家族的一员。本研究将该基因与XcmⅠ酶切的表达载体pCXSN连接,转化大肠杆菌,通过测序挑选插入正确克隆酶切验证,并成功转化农杆菌。本研究结果为进一步研究该基因的功能提供了帮助。     

白屈菜CmFAD2基因的克隆及植物转化载体构建

通过对白屈菜低温应答过程的转录组分析发现膜脂不饱和化相关基因的表达在一定过程中发生变化,脂肪酸去饱和酶基因FAD2在随温度的变化趋势为正"V"型,且表达量变化显著。利用NCBI等在线软件对序列进行相关生物学信息分析,并对白屈菜FAD家族成员FAD2基因的完整开放阅读框(ORF)进行克隆,并命名为CmFAD2。选用克隆载体pMD-19-T,转化大肠杆菌DH5α,测序验证序列正确性及完整性。将目的基因与植物表达载体pRI-201-AN连接构建重组DNA pRI-201-AN-Cm FAD2,电击法转化农杆菌LBA4404,利用菌液PCR法验证成功。该基因可作为药用植物抗寒品种创制的候选基因。     

Notch1 promotes renal tubulointerstitial fibrosis in renal tissues of diabetic mice by inhibiting autophagy through Akt/mTOR signaling pathway

AIM: To observe the changes of Notch1 expression and autophagy in the renal tissues of diabetic mice, and to explore the regulatory effect of Notch1 on tubulointerstitial fibrosis by inhibiting autophagy in diabetic nephro-pathy. METHODS: The mice were randomly divided into normal control group (db/m mice) and diabetes group (db/db mice), with 8 rats in each group. After 12 weeks of feeding, the mice were sacrificed and the corresponding biochemical indexes were measured. The protein expression of Notch1 in the renal tubular epithelial cells was observed by immunohistochemical staining. The protein levels of Notch1, PTEN, p-Akt (Thr308), Akt, p-mTOR (Ser2448), mTOR, LC3, P62, collagen type Ⅰ (Col-Ⅰ) and collagen type Ⅲ (Col-Ⅲ) were determined by Western blot. RESULTS: Compared with the db/m mice, the blood glucose, glycosylated hemoglobin, serum creatinine, triglyceride and total cholesterol were increased in the db/db mice (P<0.01). Renal tubular epithelial cell vacuolar degeneration, renal tubular expansion and interstitial inflammatory cell infiltration in db/db mouse renal tissues with HE staining were observed. The images of Masson staining showed collagenous fiber-like substance deposition in the glomerular capillaries and renal interstitium, and disarrangement of tubular structure in the renal tissues of db/db mice. The protein expression levels of PTEN and LC3-Ⅱ were decreased (P<0.01 or P<0.05), while the protein levels of Notch1, P62, p-mTOR (Ser2448), p-Akt (Thr308), Col-I and Col-III were increased in the db/db mice as compared with the db/m mice (P<0.01). However, no significant change of total mTOR and Akt proteins between the 2 groups was found. CONCLUSION: Notch1 protein expression was increased, PTEN expression was significantly reduced, Akt/mTOR pathway was activated, autophagy was inhibited, and fibrosis was aggravated in the renal tissues of the diabetic mice.     

An investigation on the effect of heat-moisture treatment on baking quality of wheat by using response surface methodology

Effect of heat-moisture treatment on quality properties of two bread wheats (cvs. Tosunbey and Bayraktar) were investigated by using response surface methodology (RSM). Temperature and moisture conditions in the experimental design were in the range of 55–95 °C and 13–19%. Heat-moisture treated grains were milled into flour and quality properties were determined. The optimum moisture-temperature combination for the highest dry gluten, Zeleny sedimentation, Alveograph W and bread volume values were estimated as 14%-63 °C for Tosunbey and 19%-55 °C for Bayraktar samples. Alveograph W seems to be a good indicator of baking quality for wheats treated at higher temperatures. In order to describe the relationship between the dependent and independent variables (moisture, temperature), the response values were fitted by second order polynomial models. Significance analysis showed that the effect of both moisture and temperature on dry gluten content, sedimentation and falling number values for Tosunbey; falling number and damaged starch values for Bayraktar were significant (p < 0.05). The effect of temperature on Farinograph water absorption, W and P/G, bread volume and firmness values were significant for both cultivars (p < 0.05). It can be concluded that improvement in baking quality can be achieved and flours with different properties can be produced by heat-moisture treatments on wheat.     

Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens

We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control.     

Zerumbone attenuates MPP+-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.     

无源蓄冷控温运输箱设计与试验

针对蓄冷运输箱信息化程度低、控温时间短、控温困难等问题,设计了一款集控温、远程监控、定位、故障诊断等功能于一体的蓄冷运输箱。以脐橙为试验对象,结合能耗模型,对蓄冷控温箱控温性能进行了研究。结果表明,箱内各截面温度不均匀系数分别为0.38、0.47、0.78,温度极差最大值为2.8℃,均匀性较好;当蓄冷剂用量为180 kg,预冷脐橙660 kg,在外部环境平均温度26.39℃的条件下,总控温时长为122 h,风机共执行控温21次,且随着蓄冷量减少,风机开启控温所用时间呈指数上升趋势,决定系数不小于0.928 0;结合能耗模型分析得出,该箱体在广州夏季高温环境下可控温5 d以上,能够满足远距离运输要求。     

热风干燥过程中山药水分状态的变化研究

为了解山药的水分赋存状态的变化，以襄阳道地山药为材料，分别在60、70、80、90、100 ℃条件下进行热风干燥，采用低场核磁共振（LF-NMR）和差式量热扫描（DSC）技术，每隔15 min测定山药在热风干燥过程中的水分状态及迁移规律。结果表明，在60~90 ℃条件下，温度越高，干基含水率的降速越快。山药的T2弛豫图谱有3个较为明显的吸收峰，随干燥进程的延续，各峰面积均明显减少，其中自由水所在峰的面积降幅最大，表明干燥过程中自由水散失最多，而且自由水逐渐向半结合水和结合水迁移，冷冻峰和解冻峰也随之变小。但在100 ℃下干燥时，样品可能因表面板结导致干基含水率、低场核磁吸收峰升高，冷冻和解冻峰面积增加。因此，在实际干燥过程中，山药的热风干燥温度不宜高至100 ℃。     

不适温度条件诱导西瓜嗜酸菌进入VBNC状态的研究

 由西瓜嗜酸菌(Acidovorax citrulli)引起的瓜类果斑病,是危害西瓜和甜瓜等葫芦科作物的一种典型的种传细菌性病害。已有文章报道西瓜嗜酸菌在铜离子诱导下可进入“有活力但不可培养”(viable but non-culturable,VBNC)状态,并在适当条件下复苏,成为生产中潜在的病害初侵染来源。本文结合实际生产中的种子温汤浸种和低温储藏等条件,分别设定了50℃、55℃高温和4℃低温对西瓜嗜酸菌AAC00-1进行处理,通过流式细胞术(flow cytometry,FCM)和平板培养法分别检测菌体活性和可培养性。结果显示,不适宜的温度条件能够诱导西瓜嗜酸菌AAC00-1进入VBNC状态,其中55℃高温20和30 min的两个处理,所有活菌均进入VBNC状态。将处理后的AAC00-1接种西瓜子叶,接种植株未表现出果斑病症状,且不能从接种部位分离得到可培养菌体。表明本诱导条件下的VBNC状态A. citrulli菌体不能通过短时间活体接种复苏。本研究分析了西瓜嗜酸菌在不适宜温度条件下进入VBNC状态的能力,为解析病害可能的初侵染来源提供了理论依据,对葫芦科作物果斑病防控具有重要的生产实践意义。