Role of calcineurin in airway remodeling in guinea pig model of asthma

AIM:To investigate the role of calcineurin (CaN) in airway remodeling in guinea pig model of asthma.METHODS:Male guinea pigs were randomly divided into three groups: control, asthma group and CsA group. The following parameters were measured: 1. The protein content, cell count and differential count of BALF; 2. The amount of [³H]-TdR incorporation into central airway smooth muscle; 3. The mean thickness of airway wall and airway smooth muscle of small airwaysl; 4.CaN activity of trachea and lung tissue.RESULTS:1. The protein content, cell count and eosinophil of BALF in CsA group were 46%, 51% and 60% lower than those in asthma group, respectively (P＜0.01); 2. [³H]-TdR incorporation in CsA group was 22% lower than that in asthma group (P＜0.05);3. The mean thickness of airway wall and airway smooth muscle were 34% and 37% less in CsA group than those in asthma group, respectively (P＜0.01); 4. CaN activity of lung tissue and trachea were 52% and 44% lower in CsA group than those in asthma group, respectively (P＜0.01).CONCLUSION:CsA reduced airway remodeling in guinea pig model of asthma, indicating the role of CaN in the airway remodeling.     

Preparation of gfp-bcl-XL-contained recombinant adenovirus vector by the homologous recombination in bacteria

AIM: To prepare gfp-bcl-X_L-contained recombinant adenovirus(rAd-gfp-bcl-X_L).METHODS: Bcl-X_L gene was amplified from pEGFP-C₃-bcl-X_L, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-X_L. Then pAdTrack-CMV-bcl-X_L was linealinzed with PmeI and co-transformed into BJ5183 bacteria with adenovirus genomic plasmid of pAdEasy-1. The identified recombinant adenovirus plasmid was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles. The target gene was detected by PCR.RESULTS: There were about 35% positive recombinant bacterial clones after the co-transformation of pAdTrack-CMV-bcl-X_L and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-X_L into 293 cells. PCR test indicated that the recombinant Ad contained bcl-X_L gene. The titer of the purified rAd-gfp-bcl-X_L was 6.5×10¹² PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-X_L. This affords a good gene transfer vector for the gene therapy in human’s diseases.     

Effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts

AIM: To investigate the effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts. METHODS: In the present experiment, the human vascular fibroblasts were cultured and effects of platelet-derived growth factor-BB on DNA and collagen protein synthesis in human vascular fibroblasts were observed by using [³H]-TdR incorporation and [³H]-proline incorporation in vitro. RESULTS: Platelet-derived growth factor-BB significantly promoted NDA synthesis and collagen protein synthesis of quiescent human vascular fibroblasts, with a maximal response at a concentration of 30μg·L^-1at 24 h and 36 h, respectively. CONCLUSION: Platelet-derived growth factor-BB promotes DNA and collagen protein synthesis in cultured human vascular fibroblasts.     

Effects of carvedilol,cilazapril and their combination on left ventricular remodeling after acute myocardial infarction in rats

AIM:To compare the effects of carvedilol, cilazapril and their combination on left ventricular remodeling(LVRM) after acute myocardial infarction(AMI) in rats. METHODS: Twenty-four hours after AMI operation, 100 surviving rats were randomly assigned to: ①AMI control(n= 25), ②AMI+carvedilol(1 mg·kg^-1 ·d^-1, n= 25)(C1), ③AMI+cilazapril(1 mg·kg^-1 ·d^-1, n= 25)(Z1), and ④ AMI+combination(n= 25) groups. Sham-operated group(n= 17) were selected randomly. After 4 weeks of therapy with the drugs gastric gavage, hemodynamic and pathological studies were performed. RESULTS: There were no significant differences in MI size among the four AMI groups(all P> 0.05) Left ventricular(LV) end diastolic pressure(LVEDP), volume(LVV), weight(LVW) and septal thickness(STh) were all higher and left ventricular pressure maximal rate of rise and fall(±d p /d t) were lower(all P< 0.01) in AMI group than sham-operated group. The LVEDP, LVV, LVW and STh were all lower and ±dp /dt were higher in Z1, C1, and combination groups than those in AMI group(P< 0.05, P< 0.01), with LVEDP and STh were more lower in the combination group than in the two monotherapy group(P< 0.05, P< 0.01), but there were no significant differences in other variables among the three therapy groups. CONCLUSION: Carvedilol, cilazapril and their combination all can prevent from LVRM after AMI in rats, improve hemodynamics and LV function, with the combination superior.     

Protein expression of cyclin D2 and p16 in proliferation and differentiation of cultured cardiac myocytes

AIM:To investigate the protein expression of cyclin D₂ and p16 in proliferation and differentiation of cultured cardiac myocytes.METHODS:One-day-old Sparague-Dawley rats were used. Cardiac myocytes(CM) were collected by a trypsin-dispersal method and cultured. Cell growth line and fluorescence activated cell sorting (FACS) were used to investigate the proliferation of CM. Ultra-thin sections were made to observe the ultrastructure of CM under transmission electron microscope. The expression of cyclin D₂ and p16 in CM were measured using immunocytochemistry and image analysis.RESULTS:①Results of cell growth line and FACS analysis showed that cultured CM could proliferate in the first 3 cultured days, but the ability decreased quickly, concomitant with differentiation. CM was obseved quiescent in cell cycle three days later. The ultrastructure of CM showed the large amount of myofilaments and mitochondrion. ②The protein expression of cyclin D₂ in 3,4,5 day CM group was 0.89 times(P＜0.05),0.80 times (P＜0.05) and 0.56 times (P＜0.01) of that in 1 day group, respectively. The expression of p16 in CM was increased during the culture process, 2,3,4,5 day group were 1.63 times, 1.72 times, 1.99 times and 2.84 times (P＜0.01) of that in 1 day group, respectively.CONCLUSION:Cultured neonatal rat cardiac myocytes could proliferate during the first 3 days after incubation, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D₂ and p16 play the key roles in CM postnatal development. Downregulation of cyclin D₂ and upregulation of p16 may induce CM differentiation.     

哈茨木霉几丁质酶诱导及其对水稻纹枯病菌的拮抗作用

 在实验室条件下分别以几丁质和水稻纹枯病菌(Rhizoctonia solani)细胞壁作唯一碳源诱导哈茨木霉(Trichoderma harzianum)菌株NF9、TC3和P1产生几丁质酶,用硫酸铵沉淀法制备几丁质酶粗提液。上述木霉菌株内切几丁质酶活性(对胶体几丁质浑浊度的减少率)分别为79.8%、74.4%和76.0%,均显著高于非诱导的阳性对照。培养第5 d几丁质诱导的木霉菌株NF9和TC3内切几丁质酶活性显著高于由水稻纹枯病菌细胞壁诱导的酶活性。体外测定表明,通过诱导的木霉菌株TC3、NF9和P1几丁质酶粗提液对水稻纹枯病病菌的拮抗圈直径可达38、21和23 mm,与非诱导的阳性对照比较有显著性差异。对木霉几丁质酶拮抗作用的特点及生防应用进行了讨论。     

三种暖季型草坪草在干旱条件下脯氨酸含量和抗氧化酶活性的变化

 测定了土壤干旱条件下矮生狗牙根、沟叶结缕草和地毯草相对含水量和相对电导率的变化。结果表明，矮生狗牙根的耐旱性较强，沟叶结缕草次之，而地毯草对干旱敏感。干旱胁迫下，地毯草积累脯氨酸最多，沟叶结缕草次之，矮生狗牙根最少，草坪草的脯氨酸的积累与耐旱性呈负相关。地毯草超氧化物歧化酶(SOD)活性持续升高，过氧化氢酶(CAT)活性在短暂升高后降低，过氧化物酶(POD)活性降低；矮生狗牙根和沟叶结缕草的SOD活性先升后降，CAT和POD活性则维持较高水平，表明草坪草的耐旱性与抗氧化酶活性有关。     

β-氨基丁酸、茉莉酸及其甲酯诱导辣椒抗TMV作用的研究

β -氨基丁酸 (BABA)、茉莉酸 (JA)及其甲酯 (MeJA)喷雾处理辣椒叶片和茎秆后可诱导辣椒获得对TMV的抗性。研究结果表明 :3种诱抗剂对不同辣椒品种所表现的诱抗效果没有显著差异 ;诱抗效果受诱抗剂的使用浓度、作物的生育期和TMV的接种浓度所影响 ;最高的诱抗效果可达到 5 0 %以上 ;BABA处理后 5～ 6d ,JA与MeJA处理后 6～ 10d接种TMV ,辣椒植株开始表达出较高的诱导抗性 ,这种抗性可持续 15d以上 ,并表现出与数量抗病性相似的特性。     

巴西蘑菇培养料筛选

对11个配方进行试验,筛选出一种适合巴西蘑菇生长的高产培养料配方:棉籽壳42％,麦草28％,牛粪20％,麦轶5％,石灰2.5％,过磷酸钙1％,石膏1％,尿素0.5％。用这种培养料栽培巴西蘑菇,产量可达到9.12～9.59 kg·m~(-2),生物学效率为39.65％～41.70％。     

小麦化感作用物的提取,分离及其对白茅的杀除效果

小麦颖壳提取物经3520树脂分离后,对所得甲醇洗脱物离心,分得油类似物、上清液及沉淀物。前两个组分在300mg/L的浓度下,对白茅生长抑制率分别为100％和98％。上清液经硅胶柱分离,得7个亚组分,其中第1、3和7号亚组分在500mg/L浓度下对白茅生长的抑制率分别为95％、91％和95％。第3亚组分经葡聚糖凝胶柱分离,又分得A、B、C 3个组分,褐色粉末状的B组分在1000mg/L浓度下使白茅的生长量下降81％。从而证明小麦颖壳对白茅的化感作用是由多个化感物共同作用的结果。室内和温室及大田的测定结果表明,小麦颖壳中的甲醇洗脱物在24mg/株的剂量下对白茅生长的抑制率可达90％以上。这暗示,小麦颖壳中的甲醇可溶物可望开发成为防治白茅的生物除草剂。