串番茄新品种红星302 的选育

红星302 是以抗病、耐低温弱光材料01-61 为母本，以01-23 为父本育成的串番茄一代杂种，无限生长类型，果实串收，果色亮红，果实椭圆形，酸甜适口，品质佳，VC 含量为364 mg · kg^-1，可滴定酸0.44%，可溶性糖4.25%，平均单果质量30.79 g，硬度为11.40×10⁵ Pa。单株结果数约为53个，产量4 900 kg ·（ 667 m²）^-1 左右，高抗蕨叶病毒病、青枯病，适宜河北地区冬季日光温室和早春保护地栽培。     

人溶菌酶密码子优化及其在牛乳腺细胞中高效表达

【目的】分析牛乳腺中7种主要乳蛋白基因密码子使用偏好性,筛选高频密码子和低频密码子,依据其对人溶菌酶基因进行密码子局部优化和全局优化,通过牛乳腺上皮细胞等多种细胞对优化效果进行分析评价,为提高重组人溶菌酶表达量,开发新型、高效、安全的重组人溶菌酶提供理论依据。【方法】利用CodonW和EMBOSS等软件对7种主要牛乳蛋白和人溶菌酶基因密码子偏好进行生物信息学分析,筛选牛乳蛋白基因高频、低频密码子并根据牛乳蛋白基因密码子使用偏好对人溶菌酶基因翻译起始区前22位密码子（LYZop22）和全局密码子(LYZop)分别进行优化。构建人溶菌酶-荧光素酶融合表达载体（pGL3-LYZcw/op22/op）和人溶菌酶过表达载体（pcDNA-LYZcw/op22/op）,将上述载体分别转染牛乳腺上皮细胞(BMEC)、牛成纤维细胞（BFFC）和C127小鼠乳腺上皮细胞等3种细胞,通过荧光素酶、实时荧光定量PCR及Western-blot等方法检测密码子优化对溶菌酶表达的影响。【结果】牛乳蛋白基因密码子偏好以GC结尾,GC3s平均含量为0.537±0.062,而人溶菌酶GC3s含量为0.407,偏好以AT结尾;聚类结果表明,牛乳中酪蛋白与乳清蛋白类基因在密码子使用偏好性上也存在一定差异。依据筛选获得的牛主要乳蛋白基因5个高频密码子（RSCU>1.5）和7个低频密码子（RSCU<0.5）对人溶菌酶基因密码子进行优化并转染多种细胞进行表达效果分析。荧光素酶分析发现,相比野生型溶菌酶密码子（LYZcw）,翻译起始区密码子优化类型LYZop22分别在BMEC、BFFC细胞中提高1.48倍（P<0.01）和1.30倍（P>0.05）;而全局密码子优化类型LYZop分别在BMEC、BFFC中提高2.2倍（P<0.01）和2.44倍（P<0.01）,说明密码子优化能明显提高人溶菌酶在多种细胞中表达量。实时荧光定量PCR结果表明,LYZop22相比LYZcw在BMEC和BFFC细胞中分别提高了2.08倍（P<0.05）和1.5倍（P>0.05）,而LYZop则分别提高了22倍（P<0.01）和17.8倍（P<0.01）,mRNA表达水平与密码子优化后的mRNA二级结构稳定性呈正相关。Western-blot结果也进一步表明,密码子优化后的LYZop22和LYZop能明显提高重组人溶菌酶在牛乳腺上皮细胞中的表达量。上述结果表明,根据牛乳腺中主要乳蛋白基因密码子使用偏好进行密码子优化,能显著提高人溶菌酶在牛乳腺上皮细胞及成纤维细胞中的表达量,且溶菌酶基因全局密码子优化效果优于翻译起始区密码子优化效果。【结论】通过生物信息学分析获得了牛乳蛋白基因使密码子使用偏好及高低频密码子;依据牛乳蛋白密码子使用偏好性对人溶菌酶密码子优化能显著提高重组人溶菌酶mRNA水平和蛋白水平的表达量,为今后利用生物反应器高效生产重组人溶菌酶奠定基础。     

不同增氧滴灌方式对蔬菜生长生理指标的影响

为了研究不同增氧方式对盆栽小白菜生长生理指标的影响,以小白菜为供试作物,采用盆栽地下滴灌的方式,以普通地下滴灌作为对照(CK),设置循环曝气(MAI)、双氧水(H₂O₂)、纯氧扩散器曝气(OC)及射流振荡器曝气(FO)4个增氧灌溉处理.结果表明,增氧地下滴灌显著提高了土壤呼吸速率,处理MAI,OC和FO较对照处理分别增大了65.87%,66.79%和111.62%.增氧地下滴灌促进了小白菜的根系生长、光合作用、蒸腾速率和气孔导度,进而提高了小白菜的物质量积累和产量.与对照相比,处理MAI的地下部鲜质量增大了42.03%,地下部干质量增大了79.85%;处理MAI,H₂O₂,OC和FO的光合速率分别增大了868.62%,794.14%,778.67%和650.19%;处理MAI,H₂O₂和OC的气孔导度较CK增大了157.14%,128.57%和85.71%,蒸腾速率增大了55.61%,32.38%和19.58%;处理MAI和H₂O₂的产量分别增大了56.36%和38.72%.综上,增氧地下滴灌可增强小白菜根区的土壤呼吸作用,改善光合作用、蒸腾速率和气孔导度,提高了产量及水分利用效率.其中,循环曝气处理的改善效果最为显著.     

Effect of pidotimod on renal function and inflammatory response in rat IgA nephropathy model

AIM:To explore the effect of pidotimod on the renal function in IgA nephropathy (IgAN) rat model, and to further study whether this effect is related to the inhibition of inflammatory response. METHODS:The SD rats (n=36) were randomly divided into control group, IgAN model group, IgAN with prednisone treatment group and IgAN with pidotimod treatment group, with 9 rats in each group. The IgAN model was induced by consecutive oral administration of bovine gamma globulin (BGG) for 8 weeks followed by injection of BGG through tail vein for 3 d. After the IgAN model was established, the drug was continuously used for 4 weeks. At the end of the treatment, the urine protein, serum creatinine and blood urea nitrogen were examined by an automated analyzer. IgA deposition in the renal tissues was observed by immunofluorescence staining. The mRNA expression levels of renal fibrosis markers transforming growth factor-β1 (TGF-β1) and fibronectin 1 in the renal tissues were detected by RT-qPCR. The mRNA and protein levels of pro-inflammatory cytokines interleukin-1β (IL-1β) and IL-6 in the renal tissues were determined by RT-qPCR and Western blot, respectively. RESULTS:No significant difference of the body weight was observed in different groups. Compared with control group, the content of urine protein, serum creatinine and blood urea nitrogen were significantly increased (P<0.01), whereas those were reversed by pidotimod treatment. The results of immunofluorescence staining showed that pidotimod inhibited IgA deposition in the IgAN rats. Pitomod treatment inhibited the mRNA expression levels of renal fibrosis markers TGF-β1 and fibronectin 1, and the mRNA and protein levels of pro-inflammatory cytokines IL-1β and IL-6 in the renal tissues of IgAN rats. CONCLUSION:Pidotimod alleviates IgAN progression in rats by inhibition of inflammatory response.     

Regulatory effect of astragaloside IV on SDF-1α/CXCR4 in EPCs

AIM: To investigate the effects of astragaloside IV (AS-IV) on chemokine receptor 4 (CXCR4) and stromal cell-derived factor 1α (SDF-1α) in endothelial progenitor cells (EPCs) and its mechanism. METHODS: Rat bone marrow-derived EPCs were cultured in vitro. The proliferation, adhesion, migration, apoptosis and tube formation capacity of EPCs treated with AS-IV and AMD3100, a specific blocker of CXCR4, were observed. The effects of AS-IV on the expression of SDF-1α/CXCR4 at mRNA and protein levels and the protein level of p-CXCR4 in the EPCs were determined. RESULTS: AS-IV significantly enhanced the proliferation, adhesion, migration and tube formation abilities of EPCs, reduced the apoptosis of EPCs, and up-regulated the mRNA and protein expression of SDF-1α and CXCR4 and the p-CXCR4 protein level in the EPCs. On the other hand, AMD3100 blocked the up-regulating effect of AS-IV on the mRNA and protein expression of CXCR4 and the p-CXCR4 protein level in the EPCs, but did not affect the effect of AS-IV on the expression of SDF-1α. CONCLUSION: AS-IV might enhance the biological function of EPCs by regulating the expression of SDF-1α/CXCR in EPCs.     

水稻资源全生育期耐盐性鉴定筛选

对来自国内外不同地区的550份水稻资源进行全生育期耐盐性鉴定。设置淡水、0.3%和0.5%盐溶液浇灌3个处理,插秧10d后,通过浇灌不同体积的淡水与海水调至设计浓度对水稻进行不同浓度的盐胁迫处理。分别调查了淡水及0.3%盐处理下水稻株高、有效分蘖数、主穗长度、主穗结实率、单株产量、抽穗期6项农艺性状和0.5%盐处理下的水稻耐盐表型。与淡水浇灌相比,全生育期在0.3%盐溶液处理下,550份水稻(100%)株高显著降低;124份(22.55%)水稻有效分蘖数(90份增加, 34份减少)、414份(75.27%)水稻主穗长度(405份变短、9份增长)、145份(26.36%)水稻主穗结实率(84份减少, 61份增加)、375份(68.18%)水稻单株产量(343份减少, 32份增加)存在(极)显著差异;水稻资源的抽穗期无显著差异。主成分分析表明,主穗结实率、有效分蘖数及单株产量3项性状累计贡献77.25%的变异。根据产量耐盐系数筛选了121份耐盐水稻资源(产量耐盐系数≥0.8),在0.5%盐胁迫持续处理42 d后,筛选了78份耐盐水稻资源(耐盐表型为3级),其中25份水稻资源全生育期在0.3%盐处理下单株产量耐盐系数≥0.8,在0.5%盐胁迫持续处理42 d后的耐盐表型为3级。筛选的耐盐水稻资源为培育耐盐新品种及深入研究耐盐机制提供材料。     

巨菌草对七彩山鸡生长性能、免疫器官以及肠绒毛形态的影响

试验旨在探究日粮中添加巨菌草(Pennisetum giganteum)对七彩山鸡(Phasianus colchicus)生长性能、免疫器官以及肠绒毛形态的影响。试验选取1周龄健康的七彩山鸡(母)210只,随机分为1个对照组(0%组)和4个饲料处理组,每组3个平行。4个饲料处理组配制时,分别添加4%、8%、12%、16%的巨菌草添加剂于基础饲料中。结果表明,与0%组相比,12%组七彩山鸡的平均体重和平均日采食量显著增加(P<0.05);4%、8%、12%组平均日增重显著大于16%组(P<0.05);8%组料重比显著降低(P<0.05);饲料处理组免疫器官指数中肝脏、脾脏指数均在0%组的基础上有所增加,但差异不显著(P>0.05);8%的巨菌草添加剂可显著增加七彩山鸡肠道的绒毛长度(P<0.05),隐窝深度显著降低(P<0.05),扩大肠道黏膜吸收面积。说明日粮中添加巨菌草可以提高七彩山鸡肠道对营养的吸收能力,一定程度上改善七彩山鸡生长性能和免疫性能。     

Effect of DNA methylation of miR-30a-5p promoter region on hepatic injury induced by homocysteine

AIM: To explore the role of DNA methylation of microRNA-30a-5p(miR-30a-5p) promoter region in hepatic injury. METHODS: Four-week-old normal mice and cystathionine β-synthase (CBS) single gene knockout mice were used and divided into normal (CBS^+/+, n=12) group and single gene knockout (CBS^+/-, n=12) group, and the mice were fed with high methionine diet for 8 weeks. HL-7702 hepatic cells were routinely cultured in vitro and divided into control group, homocysteine (Hcy) group and Hcy+5-azacytidne (AZC) group. Serum Hcy, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured by automatic biochemical analyzer. The levels of ALT and AST in the cells culture medium were determined by the microplate method. Hepatic injury in the mice were observed with HE staining. Cell viability staining was used to measure the viability of hepatocytes. RT-qPCR was used to detect the expression of miR-30a-5p in the liver tissues and hepatocytes. The correlation between the expression of miR-30a-5p and serum ALT and AST levels was analyzed by Pearson correlation analysis. DNA methylation level of miR-30a-5p promoter region in the liver tissues and hepatocytes was detected by nested landing methylation-specific PCR (nMS-PCR). RESULTS: Compared with the CBS^+/+ mice, the serum levels of Hcy, ALT and AST in the CBS^+/- mice were significantly increased (P < 0.05). HE staining showed the hepatocyte swelling and nuclear fragmentation and dissolution. The expression level of miR-30a-5p in the liver tissues was decreased (P < 0.01). Besides, the expression level of miR-30a-5p in the mice was negatively correlated with serum ALT and AST levels (r²=0.4557, P=0.0003, r²=0.4626, P=0.0003), and the DNA methylation of miR-30a-5p promoter region was increased (P < 0.01). In the HL-7702 cells, compared with control group,the ALT and AST levels were increased in Hcy group (P < 0.05, P < 0.01), and the cell viability was remarkablely decreased. DNA methylation of miR-30a-5p promoter region was increased (P < 0.01), which decreased after treated the cells with AZC (P < 0.05), while the expression level of miR-30a-5p in the cells was increased (P < 0.05). CONCLUSION: Hypermethylation of miR-30a-5p promoter region may play an important role in hepatic injury.     

不同光质照射下真姬菇的生长发育与光受体的响应表达

采用 5 种不同光质光源，分别对真姬菇（Hypsizygus marmoreus）的菌丝、原基和子实体进行照射，检测菌丝生长速度、原基形成数量、子实体菌柄长度、菌柄直径、菌盖直径以及产量等。结果显示：真姬菇菌丝在黑暗和红光下生长较快，在红绿蓝复合光下生长最慢；在蓝光下形成的原基数量最多，而在红光下原基数量最少；子实体阶段，黑暗下菌柄的长度及直径最大，绿光下菌柄长度及直径最小；而在蓝光和红绿蓝复合光下菌盖直径最大。相同培养时间，真姬菇在黑暗和蓝光下产量较高，在绿光下产量最低。进一步检测真姬菇光受体基因 WC1 和 WC2 在不同光质条件下的差异表达。结果表明，在菌柄中 WC1 在黑暗下表达量最高，WC2 在蓝光下表达量最高；而在菌盖中 WC1 和 WC2 均在蓝光下表达量最高。     

LED补光组合对大棚越橘生长发育的影响

以2年生南高丛越橘‘Emerald’为材料，以大棚内自然光作为对照，研究LED光源的红蓝光（3︰1和6︰1）、紫外光（UVA）对植株长势、叶片光合作用、碳氮代谢、开花基因表达及开花率、果实品质的影响。结果表明：红蓝光组合处理下‘Emerald’的植株营养生长较旺盛，株高、1年生枝条长度和粗度显著高于对照。此外，红蓝光（6︰1）处理下叶片的叶绿素相对含量、比叶重和净光合速率均显著提高。红蓝光组合也可诱导植株开花，开花基因FT表达量和开花率明显高于对照。紫外光下叶片的氮含量、开花率及FT基因表达量显著高于对照。不同光质组合补光对‘Emerald’的果实品质有显著影响，红蓝光（3︰1）处理下果实的质量、横纵径、可溶性固形物、可溶性糖、花青素含量和糖酸比均高于对照。总之，不同光质补光会促进越橘‘Emerald’的生长发育，红蓝光6︰1组合对促进营养生长作用相对较大，红蓝光3︰1组合对提高果实品质效果较好。紫外光虽能改变植株形态和促进开花，但果实品质提高效果较红蓝光处理稍弱。