Quick detection of sex determining region protein gene in mouse fetuses

AIM: To detect quickly the Y-chromosome specific sex determining region protein (Sry) gene in mouse fetuses on embryonic day 14.5 with a PCR method. METHODS: We designed specific primers with the OLIGO 5. 0 software. Templates were prepared in 30 minutes by the following way. About 1 mg embryonic tissue but not fetal liver was suspended, and treated with 200μL of lysis buffer, consisting of PCR buffer containing 20 mg/L proteinase K, 0. 5% NP-40, and 0.05% Tween 40, at 60°C for 15 minutes, heated for 5 minutes at 100 °C, 10μL was used as template. The PCR react ion was performed in 50μL, using two sets of primers specific for Sry gene (chromosome Y) and IL-3 gene (chromosome 11) . PCR conditions and cycle numbers were optimized. The assessment of the results was done by electrophoresis in 3% agarose run at high voltage. The specificity of the method was conf irmed by fluorescent in situ hybridization (FISH) using a specific male probe on embryonic tissue cells. RESULTS: Electrophoresis showed that PCR product of male control DNA consisted of a 649 bp product representing the IL-3 gene and a 444 bp product representing the Y-specific Sry gene, female control DNA only one 649 bp product. Fetuses with two bands matching those as seen inmale control DNA are the presumpt ive male fetuses. Fetuses, only the IL-3-associated 649 bp band, are the presumptive female fetuses. These were confirmed by FISH. The ent ire procedure took <3. 5 h. CONCLUSION: The established PCR assay offers a quick, simple, accurate, and sensitive detection of sex determining region protein gene in mouse fetuses. This method allowed the preparation and culture of pure male and female hematopoietic stem cells from fetal tissue.     

Bcl-2 antisense oligodeoxynucleotide increases the sensitivity of HL60 and K562 cells to daunorubicin

AIM:To investigate whether the bcl-2 antisense oligonucleotide increases the sensitivity of HL60 and K562 cell lines to daunorubicin.METHODS:IC50 for HL60 and K562 was determined with MTT method, the expression levels of Bcl-2 protein were assayed by immunofluorescence using fluoresce isothiocyanate labeling. In addition, apoptosis was detected by morphological observation and flow cytometric analysis of DNA fragmentation.RESULTS:It was found that the two oligonucleotides directed against the coding region and the translation initiation of bcl-2 mRNA, combined respectively with daunorubicin, inhibited expression of bcl-2 protein, increased apoptosis in HL60 and K562 cells, and decreased IC50 of daunorubicin significantly (P＜0.05). Compared to the antisense oligonucleotide directed against the translation initiation of bcl-2 mRNA, the antisense oligonucleotide directed against the coding region showed stronger effects in the aspects of increasing the sensitivity of HL60 cells to daunorubicin (P＜0.05).CONCLUSIONS:These two antisense sequences in the translation initiation and the coding region of bcl-2 mRNA increased the sensitivity of HL60 and K562 cell lines to daunorubicin in a sequence-specific manner.     

Rat urotensin-II-induced main pulmonary artery contractions are mediated by MAPK

AIM: To investigate rat Urotensin-II(rat U-II)-induced vasoconstriction of rat main pulmonary arteries and the role of mitogen-activated protein kinase(MAPK). METHODS: The main pulmonary artery was dissected from the male Sprague-Dawley rats and artery ring width was 3-4 mm. Concentration-response curves were generated to rat U-II(0.03 nmol/L-30 nmol/L).Inhibitor of MAPK, PD 98059(0.1 μmol/L-10 μmol/L) were added into the medium after rat U-II(30 nmol/L)induced vasoconstriction had reached plateau to construct the relaxant concentration-response curves and their EC₅₀ and E_max. RESULTS：Rat U-II was a potent vasoconstrictor of isolated rat main pulmonary arteries [EC₅₀=7.95±0.40, E_max=(14.28±6.34)% of the response to 60 mmol/L KCl]; PD 98059 caused concentration-dependent relaxations of rat U-II precontracted arteries [EC₅₀=5.91±0.45, E_max=(81.39±13.65)%]. CONCLUSION: Rat U-II was a potent vasoconstrictor of rat main pulmonary arteries and this response was mediated through MAPK.     

Using laser to measure stem thickness and cut weed stems

Summary Stem thickness of the weed Solanum nigrum and the crop sugarbeet was determined with a He–Ne laser using a novel non‐destructive technique measuring stem shadow. Thereafter, the stems were cut close to the soil surface with a CO₂ laser. Treatments were carried out on pot plants, grown in the greenhouse, at two different growth stages, and plant dry matter was measured 2–5 weeks after treatment. The relationship between plant dry weight and laser energy was analysed using two different non‐linear dose–response regression models; one model included stem thickness as a variable, the other did not. A binary model was also tested. The non‐linear model incorporating stem thickness described the data best, indicating that it would be possible to optimize laser cutting by measuring stem thickness before cutting. The general tendency was that more energy was needed the thicker the stem. Energy uses on a field scale are discussed.     

涩柿脱涩保脆研究初报

研究了常温条件（14－19℃）和低温条件（－1－1℃）下，不同容器和不同二氧化碳（CO2）浓度对柿果脱涩保脆的影响。试验表明，低温条件下，采用密封性好的容器，保持80％CO2，脱涩时间虽稍长，但硬果率可达到96％以上，果实硬度达10kg/cm^2以上，脱涩保脆效果最好。     

根癌农杆菌介导绿色荧光蛋白基因转化印度酸桔的研究

 通过根癌农杆菌介导将绿色荧光蛋白基因转入印度酸桔的胚性愈伤组织中, 经潮霉素筛选,获得抗性愈伤组织, 并再生植株。对这些植株进行GUS 染色、PCR 分析、绿色荧光检测和Sourthern 杂交验证, 结果表明绿色荧光蛋白已经在转基因植株中表达。     

几种落叶果树H2O2含量变化与自然休眠关系的研究

 以设施栽培中常见的几种核果类果树品种和两个葡萄品种为试材, 分析了芽休眠期间 H₂O₂含量变化动态, 并探讨了温度、生长调节剂及化学破眠物质对H₂O₂含量影响的效应。结果表明: 休眠期间,不同树种( 品种) 芽内 H₂O₂含量存在差异, 基本趋势是晚熟品种高于早熟品种, 花芽高于叶芽, 但葡萄品种相反, 早熟的‘京秀’高于晚熟的‘巨峰’; 休眠期芽内 H₂O₂含量基本呈稳步上升后急剧下降的趋势,不同品种急剧下降的时间略有差别, 且与自然休眠解除的时间相吻合。低温(5 ℃) 处理显著增加了芽中 H₂O₂含量, 中温(10 ℃) 使 H₂O₂含量略有增加, 而高温(20 ℃) 却导致 H₂O₂含量降低。休眠前期50 mg.L^-1 ABA 处理显著提高了芽中H₂O₂含量, 而100 mg.L^-1的GA₃和6-BA 处理有减少 H₂O₂含量的趋势, 但二者差异不明显。热带地区常用的化学破眠物质对芽 H₂O₂的影响因树种( 品种) 、使用时期不同而异, 硫脲、KNO₃前期使用对核果类果树影响明显, CaCN₂对核果类无明显效应, 但对葡萄品种作用显著。果树芽 H₂O₂含量的动态变化表明, H₂O₂可能是低温解除自然休眠的原因。     

日光温室黄瓜栽培CO_2浓度的消长规律初探

近 3a(年 )的研究结果表明 :日光温室内CO2 浓度有明显的季节变化和日变化。在整个生长期内因通风时间和通风量的不同 ,日光温室内的CO2 浓度 11月和 3月较高 ,5月较低。各时期日变化基本相同 ,但变化幅度因季节而异 ,上午随Pn的逐渐增大而下降 ,中午 12 :0 0～ 14 :0 0时降至最低 ,下午又随Pn的减小而缓慢回升。叶片的光合作用、呼吸作用和土壤呼吸是影响日光温室内CO2 浓度日变化的主要因素。有机肥施用量对室内土壤呼吸和日光温室CO2 浓度有较大影响 ,在有机肥充足的条件下 ,室内CO2 浓度基本满足黄瓜光合作用的需要 ,无须补施 ,如果在作物生长期间再定时随水向土壤中冲施有机肥 ,效果就更好     

A reactive pattern of ischemia-induced nestin protein in the rat brain

AIM:To explore the expressive profile of nestin protein in the focal ischemic brain and to study the recovery mechanism of brain focal infarct.METHODS:Cellular morphology,time-course and distribution pattern of nestin positive response were immunohistochemically examined in different brain regions of 36 adult male SD rats. RESULTS:Nestin positive response of different brain regions in sham operated rats was present in small- and micro-vasculartures and the third ventricle bottom and ependyma. A large number of nestin positive cells were detected in ischemic brain, and were more remarkable in the cortical areas of parietal lobe and preoptic area as well as ischemic caudoputamen. Stellate nestin positive cells were located in the deep layer of ischemic cortex, but fibrillary cells were located in the shallow layer. Nestin positive cells in the ischemic caudoputamen showed the same changes of morphology as those cells in the deep layer of ischemic cortex. Morphological and number alterations of nestin positive cells were the most remarkable at 1 weeks post-ischemia, which showed more hypertrophy and proliferation in morphology, and a marked increase in number was present in the ischemic cerebral cortex and the ischemic caudoputamen. These alterations of nestin positive cells persisted up to 6 weeks post-ischemia, and then, the nestin positive response in the ischemic brain decreased gradually.CONCLUSION:Focal cerebral ischemia induces nestin re-expression on reactive astrocytes, which may be very important to the self-recovery of cerebral infarct.     

钙、萘乙酸对亚精胺在猕猴桃贮藏期间作用效果的影响

以5年生中华猕猴桃品系(湘源81-2)为试材,进行了果实采收后Spd(亚精胺)、Spd+Ca2+、Spd+NAA浸果处理,研究钙、萘乙酸对多胺作用效果的影响。结果表明:不同处理均不同程度地抑制了果实的呼吸强度、果肉PG活性和细胞膜透性的增加,其中Spd+NAA浸果处理的作用效果最为显著,贮藏68d时,果肉硬度最高(0.259MPa),软果率较低(79%),好果率达100%;其次为Spd处理。