Anti-inflammatory potential of black cumin seed oil and its nanoemulsion formulation against lipopolysaccharide-induced liver injury in mice

Objective:To evaluate the anti-inflammatory effect of black cumin seed oil(BCSO)and its nanoemulsion on lipopolysaccharide(LPS)-induced liver injury in mice.Methods:LPS-induced acute liver injury mouse model was used to evaluate the effects of BCSO and its nanoemulsion formulation on liver function.Hepatic inflammatory markers including Toll-like receptor 4(TLR4),interleukin(IL)-1β,heme-oxygenase 1,BAX,and BCL-2 were assessed using real-time PCR.Additionally,protein levels of reduced glutathione,tumor necrosis factor-α,and IL-6 were measured using ELISA,and histological analysis was performed.Indomethacin was used as a standard positive control for comparison.Results:BCSO reduced LPS-induced liver injury and exhibited strong anti-inflammatory effects by downregulating the expression of TLR4,IL-1β,IL-6,tumor necrosis factor-α,and heme-oxygenase 1.Additionally,BCSO demonstrated antioxidant properties by increasing reduced glutathione protein levels and decreasing key apoptotic markers BAX and BCL-2 in hepatocytes.The nanoemulsion formulation further enhanced these anti-inflammatory,antioxidant,and anti-apoptotic effects,and histological examination confirmed this effect.Combining BCSO with indomethacin at a lower dose improved efficacy,thereby reducing its potential side effects.Conclusions:The investigation reveals the anti-inflammatory impact of BCSO and its nanoemulsion formulation on LPS-induced liver oxidative stress,inflammation,and apoptosis.

Genetic Variability and Phenotypic Correlations Study among Grain Quality Traits and Mineral Elements Concentrations in Colored and Non-Colored Rice (Oryza sativa L.)

Twenty-four rice genotypes were examined to assess genetic variability,heritability,and correlations for seven-grain quality traits,eight nutritional elements,and protein.ANOVA revealed significant differences for the quality traits studied.For every trait under study,the phenotypic coefficient of variation was higher than the correspon-dence genotypic coefficient of variation.Heritability in a broad sense varied from 29.75%for grain length to 98.31%for the elongation trait.Hulling percentage recovery had a highly significant positive correlation with milling and head rice percentage.Consequently,milling percentage had a highly positive correlation with head rice percentage.In amylose percentage,all the genotypes belonged to low amylose except the Hassawi-1 variety,which had intermediate amylose content.Mineral nutrition contents of magnesium(Mg),sodium(Na),potas-sium(K),calcium(Ca),copper(Cu),manganese(Mn),zinc(Zn),iron(Fe),or protein percentage gave different variations for 24 rice genotypes under all the nutritional elements.Among the 24 genotypes,ten rice genotypes–HighNutrient-1,HighNutrient-2,HighNutrient-9,HighNutrient-8,HighNutrient-3,Hassawi-2,HighNutrient-7,HighNutrient-6,Hassawi-1,and HighNutrient-4–had the highest heist value for all nutritional and protein con-tents,and could be used as a donor to improving new varieties.There was a positive and significant correlation between magnesium Mg,K,Zn and Fe.Consequently,K had a positive correlation with zinc Zn,Fe,and protein percentage.Clustering analysis was divided into two groups:thefirst group included all genotypes rich in nutri-ents,while the remaining genotypes with low nutritional content were included in the second group.

Trimethylamine N-oxide generation process was influenced by the proportion and source of macronutrients in the diet

Trimethylamine N-oxide(TMAO)is a risk factor of various chronic diseases,which was produced by metabolism from precursors to trimethylamine(TMA)in gut and the oxidation from TMA in liver.The TMA generation was influenced by diet,mainly due to the rich TMAO precursors in diet.However,it was still unclear that the effects of different proportion and source of macronutrients in different dietary pattern on the production process of TMAO.Here,the generation of TMA from precursors and TMAO from TMA was determined after single oral choline chloride and intraperitoneal injection TMA,respectively,in mice fed with carbohydrates,proteins and fats in different proportion and sources.The results suggested that the generation of TMAO was increased by low non-meat protein and high fat via enhancing the production of TMAO from TMA,and decreased by plant protein and refined sugar via reducing TMA production from precursors in gut and TMAO transformation from TMA in liver.The high fat and high sugar diets accelerating the development of atherosclerosis did not increase the production of TMAO,the risk factor for atherosclerosis,which indicated that the dietary compositions rather than the elevated TMAO level might be a more key risk factor for atherosclerosis.

The role of polymorphic cytochrome P450 gene(CYP2B6)in B-chronic lymphocytic leukemia(B-CLL)incidence and outcome among Egyptian patients

Cytochromes P450(CYPs)play a prominent role in catalyzing phase I xenobiotic biotransformation and account for about 75%of the total metabolism of commercially available drugs,including chemotherapeutics.The gene expression and enzyme activity of CYPs are variable between individuals,which subsequently leads to different patterns of susceptibility to carcinogenesis by genotoxic xenobiotics,as well as differences in the efficacy and toxicity of clinically used drugs.This research aimed to examine the presence of the CYP2B6*9 polymorphism and its possible association with the incidence of B-CLL in Egyptian patients,as well as the clinical outcome after receiving cyclophosphamide chemotherapy.DNA was isolated from whole blood samples of 100 de novo B-CLL cases and also from 100 sex-and age-matched healthy individuals.The presence of the CYP2B6*9(G516T)polymorphism was examined by PCR-based allele specific amplification(ASA).Patients were further indicated for receiving chemotherapy,and then they were followed up.The CYP2B6*9 variant indicated a statistically significant higher risk of B-CLL under different genetic models,comprising allelic(T-allele vs.G-allele,OR=4.8,p<0.001)and dominant(GT+TT vs.GG,OR=5.4,p<0.001)models.Following cyclophosphamide chemotherapy,we found that the patients with variant genotypes(GT+TT)were less likely to achieve remission compared to those with the wild-type genotype(GG),with a response percentage of(37.5%vs.83%,respectively).In conclusion,our findings showed that the CYP2B6*9(G516T)polymorphism is associated with B-CLL susceptibility among Egyptian patients.This variant greatly affected the clinical outcome and can serve as a good therapeutic marker in predicting response to cyclophosphamide treatment.

Physico-Chemical and Nutritional Properties of a Food Broth Based on Nere (Parkia biglobosa)

Purpose: Diet and eating habits are major risk factors for the health and the development of disease, such as, for example, metabolic disorder leading to cardiovascular pathology and cancer, decreased immunity exposing to infections. This study of the physico-chemical and nutritional properties of a soumara-based food broth was carried out with the aim of promoting the consumption of organic broth made from nere seeds (soumara). That is to alleviate certain metabolic diseases, which is a matter of food safety, and also to limit the risk for the health about the consumption of some cooking stocks on the market. Methods: Several natural ingredients such as nere seeds (soumara), ginger, black pepper, parsley and garlic were used to create a nere-based stock. All these ingredients were freeze-dried and the powder obtained was used to make the broth, regarding their physical and chemical properties. Results: The broth had a good protein content of 17.41 ± 0.367 g/100g, a lipid content of 16.80 ± 0.08 g/100g and a fiber content of 8.66 ± 0.04 g/100g. In terms of nutritional values, the broth showed good levels of calcium 184.21 ± 0.09 mg/100g, potassium 50.04 ± 1.45 mg/100g and iron and zinc. In terms of antioxidant activity, the broth also showed good antioxidant activity. Conclusion: Regarding the properties of our food broth, whose composition is based on natural ingredients, could be recommended for consumption and, its properties, could play an important role in preventing and combating certain metabolic diseases.

Human chemerin induces eryptosis at concentrations exceeding circulating levels

Introduction:Human chemerin is an adipokine that regulates chemotaxis,inflammation,and glucose metabolism.In addition,accumulating evidence suggests that chemerin promotes apoptosis,autophagy,and pyroptosis.However,there are no data on its impact on eryptosis.The current study aimed to analyze the effects of human active Glu^(21)-Ser^(157) chemerin on eryptosis in vitro.Materials and Methods:Human chemerin 0-2-10-50μg/mL was incubated for 24 h with human erythrocytes(hematocrit 0.4%)obtained from eight healthy individuals.Flow cytometry-based determination of phospholipid scrambling,reactive oxygen species(ROS)production,and intracellular Ca^(2+)levels was performed.To supplement data on ROS and Ca^(2+)signaling in chemerin-mediated eryptosis,incubation in the presence or absence of antioxidants vitamin C and N-acetylcysteine and Ca^(2+)-binding agent EGTA was carried out,respectively.Confocal microscopy-based techniques were used to detect reactive nitrogen species(RNS)generation,involvement of caspase-3 and caspase-8,as well as the state of lipid order in cell membranes of erythrocytes exposed to human Glu^(21)-Ser^(157) chemerin.Results:Our observations suggest that human Glu^(21)-Ser^(157) chemerin had no impact on eryptosis parameters at 2μg/mL.However,chemerin stimulated phosphatidylserine externalization,ROS production,and Ca^(2+)accumulation at higher concentrations suggesting activation of eryptosis.Ca^(2+)uptake turned out to be at least partly required for chemerin-mediated eryptosis.Chemerin-mediated erythrotoxicity was additionally mediated by RNS,caspase-3,and caspase-8.Moreover,Glu^(21)-Ser^(157) chemerin promoted reduction in the liquid-ordered phase of cell membranes in erythrocytes.Conclusions:The present study first discloses that human chemerin can induce eryptosis via Ca^(2+)-dependent mechanisms at concentrations noticeably exceeding circulating levels.Thus,chemerin-induced eryptosis can hardly contribute to eryptosis-mediated anemia in diseases associated with enhanced levels of chemerin in blood.

Effects of Paclobutrazol Seed Priming on Seedlings Quality,Physiological and Bakanae Disease Index Characteristics of Rice(Oryza sativa L.)

Rice(Oryza sativa L.)is one of the most important cereal crops in the world.Bakanae disease is a significant rice disease widely distributed in rice-growing regions worldwide.Therefore,the present investigation aimed to assess the optimal concentrations of paclobutrazol(PBZ)as a treatment for rice grains(cv.Sakha 108)to control bakanae disease,also evaluating its impact on grain germination,seedling growth parameters as well as disease index.Paclobutrazol concentrations had no significant impact on seed germination,regardless of whether the seeds were incubated with Fusarium fujikuroi or not.Application of PBZ,either alone or in combination with fungal pathogens,negatively impacted the rice seedlings’height.Paclobutrazol at 25,50 and 100 mg/L,combined with the fungal pathogen positively impacted root length.Paclobutrazol at 3 and 6 mg/L mitigated the adverse impact on chlorophyll pigments content in infected seedlings.The highest proline contents were achieved by 100 mg/L PBZ alone or in combination with fungal pathogens.It has been observed that the application of PBZ,either alone or in combination with a fungal pathogen,leads to the enhancement of catalase,peroxidase,and polyphenol oxidase activities.The median lethal concentration of PBZ was 0.874 mg/L;applying low concentrations of paclobutrazol effectively increased the percentage of fungal growth suppression.Application of PBZ,at higher concentrations(50 and 100 mg/L),decreased infection percentage and disease severity index(DSI)significantly.These findings suggest that PBZ can be an effective treatment for controlling bakanae disease and enhancing resistance in rice plants.

Neurocognitive Profile of People Living with Human Immunodeficiency Virus Initiating Treatment in Kinshasa in the Dolutegravir Era, Democratic Republic of Congo

Context: Antiretroviral therapies improve the prognosis of NeuroAIDS contrasting with a high frequency of Minor Neurocognitive Disorders (MND) even in aviremic subjects. Objective: The objective of this study is to present the neurological and cognitive profile of People Living with HIV (PLHIV) initiating antiretroviral treatment in Kinshasa in the era of Dolutegravir (DTG). Methods: This is a multicenter, cross-sectional study with a descriptive aim carried out in 16 HIV Outpatient Treatment Centers (OTC) in Kinshasa from October 4, 2021 to February 15, 2022. The International HIV Dementia Scale (IHDS) correlated with the Activities of Daily Living (IADL) scale facilitated the categorization of NeuroCognitive Disorders (NCD) of PLHIV evaluated after carrying out a summary neurological examination. Results: Of the 96 patients recruited, 56.3% were women with a sex ratio of 0.68. The average age was 40.1 ± 12.1 years. The secondary education level was the majority at 64.6%. Malaria (44.8%) and tuberculosis (32.3%) were more common as opportunistic infections. They were alcoholics in (30.2%). Their history was heart disease (15.6%), high blood pressure (18.8%);drug abuse (10.4%). The IHDS score was light in 55.2% of cases. The correlation between IHDS/IADL watches asymptomatic neurocognitive impairments (ANI) in 77.1%, almost all of subjects are found with normal overall functioning (94.8%) and a disturbed neurological examination in 53.1% of cases with a predominance of motor impairments in 79.1%. Conclusion: In view of these results, early and systematic screening of NCD and associated factors remains necessary in our context. .

Neuralgic amyotrophy with hourglass-like constrictions:A case report

BACKGROUND Neuralgic amyotrophy(NA)is a rare disease with sudden upper limb pain followed by affected muscle weakness.The most commonly affected area in NA is the upper part of the brachial plexus,and the paraspinal muscles are rarely affected(1.5%),making these cases difficult to distinguish from cervical radiculopathy.CASE SUMMARY A 76-year-old male presented to the emergency department with left hip pain post-fall.After undergoing left femoral neck fracture surgery,he experienced sudden left shoulder pain for 10 days with subsequent left arm weakness.Cervical spine computed tomography revealed mild right asymmetric intervertebral disc bulging with a decreased C5-6disc space.Three weeks later,an electrodiagnostic study confirmed brachial plexopathy findings involving the cervical root.Magnetic resonance neurography was performed for a differential diagnosis.Contrast enhancement was identified at the upper trunk of the brachial plexus,including the C5 nerve root.A suprascapular nerve hourglass-like focal constriction(HLFC)was also identified,confirming NA.After being diagnosed with NA,the patient received 15 mg prednisolone,twice daily,for 3 weeks.Physical therapy was initiated,including left arm strengthening exercises and electrical stimulation therapy.Left shoulder muscle strength significantly improved one CONCLUSION NA's unique features like HLFC and paraspinal involvement are crucial for accurate diagnosis,avoiding confusion with cervical radiculopathy.

Soil Physico-Chemical Properties and Different Altitudes Affect Arbuscular Mycorrhizal Fungi Abundance and Colonization in Cacao Plantations of Cameroon

This study aims to investigate the abundance of AMF according to soil properties and altitudes in different cacao plantations of Cameroon. Physico-chemical analyses were made on soil samples collected from three agro-ecological zones. Soil samples were also used to evaluate directly the AMF abundance following the various altitudes and after trapping by sorghum plant. The results showed that soil properties, AMF spore abundances and colonization fluctuated significantly at different altitudes. The most represented texture was sandy loam. The bimodal zone presented a homogeneous texture (sandy loam) in all its localities. Cacao soil chemical characteristics showed that, the highest nitrogen rate (0.47%;p 0.05, Scott-Knott test) was recorded at Melong in a monomodal zone while Tonga in the Western highlands displayed the lowest rate (0.13%). Soil P concentration was significantly high in monomodal zones (Mbanga and Melong). Soil pH level indicated that the soil from Tonga in the Western highlands was neutral (pH = 6.67), and soils of other localities under study were acidic with the lowest (4.75) pH level recorded at Melong in a monomodal zone. In soil samples, the highest spore density (1.03 spores/g soil) was observed at Ntui in Bimodal zone, while the lowest spore density (0.26 spores/g soil) was observed at Bafang in the Western highlands. Root colonization showed that the sample from Bokito in a bimodal zone displayed the best frequency of mycorrhization (86.11%) while the sample from Bafang in the Western highlands recorded the lowest (27.11%). The PCA analysis highlighted that available phosphorus, pH and altitude all strongly correlated with AMF root colonization ability and can be used as a predictor of AMF colonization ability in cacao rhizosphere.

Total Fat, Fatty Acid Composition, Tocopherol and Tocotrienol Profiles in Fermented Beans of Ten Controlled Pollinated Cocoa (Theobroma cacao L.) Hybrids from Cameroon

This study aimed to discriminate ten Cameroonian cocoa hybrids according to their total fat, fatty acid composition, tocopherol and tocotrienol profiles. Six cocoa clones from the gene banks of the Cameroon Cocoa Development Corporation were used to create hybrids. The determination of fatty acid composition was carried out by using a gas chromatography (GC) apparatus coupled by a flame ion detector (FID). Tocopherol and tocotrienol analysis was performed by upper high-performance liquid chromatography (UHPLC). Information on the impact of the genotype on the cocoa fat composition was provided. The major fatty acids (FA) in fermented samples are stearic (34.57%), palmitic (26.13%), oleic (34.13%) and linoleic (3.16%) acids. (35.05% to 35.6%). SCA12 × ICS40, SCA12 × SNK13, SNK13 × T79/501 have the least hard cocoa butters. Tocopherols analysis showed a predominance of γ-tocopherols (94.64 ± 1.51 to 292.16 ± 3.17 µg∙g−1), whereas only a small amount of β and δ-tocopherol (from 0.46 to 2.78 µg∙g−1 and 0.12 to 5.82 respectively) was observed. No γ-tocotrienol was found in fermented samples. A differentiation in terms of total fat and tocopherol content was observed amongst hybrids with the same mother-clone, suggesting an impact of pollen on these compounds.

Strawberry (Fragaria spp.): Cultivation, Production, Consumption, and Marketing in Cameroon

Strawberry (Fragaria spp.) is one of the most important fruits classified as exotic fruits imported into Cameroon. To have an inventory of its cultivation in Cameroon, a survey study was carried out among eight farms of Fragaria spp. from January 2021 to February 2022. The plant was introduced in Cameroon in 2018. There are 13 varieties of Fragaria spp. currently cultivated. Among these 13 varieties, eleven are hybrids of Fragaria x ananassa (“Amiga”, “Amine”, “Camarosa”, “Chandler”, “Charlotte”, “Elsanta”, “Gariguette”, “Madame Moutot”, “Ostara”, “Ruby gem” and “San Andreas”), and two of the hybrids of Fragaria vesca (“Maestro” and “Mara des bois”). The cropping system, irrigation system, and type of fertilizers applied differ from one strawberry farm to another. Biofertilizers (such as mycorrhizal), inorganic and organic fertilizers are actually used to improve production. The potential annual production of strawberries from January 2021 to February 2022, estimated based on the survey data, was 21.216 tons for all growers. Among these eight production farms, the Lolodorf BIO Farm presents 6000 kg (six tons) of strawberries and 100,000 stolons (seedlings) produced, from seven varieties of Fragaria spp. cultivated, with 6 varieties which are hybrids variety Fragaria x ananassa (“Amiga”, “Amine”, “Chandler”, “Gariguette”, “Madame Moutot”, and “Ruby gem”), and one which is a hybrid of Fragaria vesca (“Mara des bois”). Certain diseases were also observed and recorded depending on the growing areas.

黑曲霉葡萄糖氧化酶基因的克隆及其在酵母中的高效表达

将黑曲霉葡萄糖氧化酶 (GOD)基因重组进大肠杆菌 酵母穿梭质粒pPIC9,转化甲基营养酵母Pichiapasto risGS115 ,构建出GOD的高产酵母工程菌株。在酵母α Factor及AOX1基因启动子和终止信号的调控下 ,黑曲霉GOD在甲基酵母中大量表达并分泌至胞外 ,经甲醇诱导 3～ 4d ,发酵液中的GOD活力可达 30～ 40u mL。SDS PAGE证实GOD在培养物上清中的含量显著高于其它杂蛋白 ,约占胞外蛋白总量的 6 0 %～ 70 % ,经QSepharoseTM FastFlow离子交换柱一步纯化即达电泳纯。重组酵母GOD比活达 42 6 6 3u mg蛋白 ,是商品黑曲霉GOD的 1 6倍。动力学性质分析表明 ,重组酵母GOD的Km 和kcat分别为 38 2 5mmol L和 34 92 6 6s- 1 ,与商品黑曲霉GOD相比 ,具有更高的催化效率。重组酵母GOD的高活力特性可有效提高葡萄糖传感器的线性检测范围。

大肠杆菌碱性磷酸酶的体外定向进化研究

大肠杆菌碱性磷酸酶 (E .colialkalinephosphatase,EAP ,EC 3 1 3 1)是一个非特异性二聚体磷酸单酯酶 .采用易错聚合酶链反应 (errorpronePCR)的方法 ,在原有高活力突变株的基础上 ,对EAP远离活性中心催化三联体的区域进行定向进化 ,经两轮errorpronePCR ,获得催化活力较亲本D10 1S突变株提高 3倍、较野生型酶提高 35倍的进化酶 4 186 ,并对该酶的催化动力学特征进行了分析 .进化酶基因的DNA测序表明 4 186含两个有义氨基酸置换 :K16 7R和S374C ,二者既不位于底物结合位点 ,也不位于酶的金属离子结合位点 .

云南恶性疟原虫氯喹抗药性基因76位点突变研究

目的研究云南恶性疟原虫氯喹抗药性基因(pfcrt)76位点突变的情况,以及与抗药性表现型的关系。方法应用PCR和限制性酶切片段长度分析方法,检测现症病人干滤纸血样的恶性疟原虫pfcrt基因点突变。结果云南省恶性疟原虫pfcrt基因76位点的突变型很高,占85.0%(51/60);野生型和混合型较少,分别占8.3%(5/60)和6.7%(4/60)。体内法测定的氯喹抗性和敏感样本均有pfcrt76突变型;体外法测定的17份氯喹抗性样本中,有13份带有pf-crt76突变型。结论云南省恶性疟原虫pfcrt基因氨基酸编码76位点突变频度很高。体内和体外法测定的氯喹抗性表现与pfcrt76突变型有较高的一致性。

NK4基因转染对裸鼠胰腺癌移植瘤生长的影响

背景与目的:NK4不仅是肝细胞生长因子的拮抗剂,而且是血管形成的抑制剂。研究已经证实NK4可以抑制肿瘤的生长和转移,但有关其在胰腺癌中的作用,目前少见文献报道。为此,本研究旨在探讨NK4基因在裸鼠体内的抗胰腺癌作用及其可能的机制。方法:建立裸鼠胰腺癌皮下移植瘤模型,构建NK4基因真核细胞表达载体并转染入瘤体内,转染前后称其体重、瘤重和测肿瘤体积,采用免疫组织化学和脱氧核糖核酸末端转移酶介导的缺口末端标记技术对裸鼠胰腺癌组织中的凋亡细胞、增殖抗原和微血管密度进行观察和比较。结果:4周后,NK4转染组裸鼠移植瘤体积为(1.39±0.33)cm3,明显小于对照组和空载体组[(2.06±0.55)cm3和(1.90±0.36)cm3,P<0.01];其瘤重为(1.30±0.81)g,也显著低于对照组和空载体组[(3.45±1.88)g和(3.14±1.51)g,P<0.01],抑瘤率为62.29%。NK4转染组肿瘤细胞的凋亡指数为9.34±0.91,显著高于对照组和空载体组(4.13±0.79和3.94±1.03,P<0.001);而NK4转染组肿瘤细胞的增殖指数为53.88±4.30,与对照组间和空载体组比较无显著性差异(56.24±4.03和54.33±5.41,P>0.05);NK4转染组肿瘤组织的微血管密度为(12.24±4.63),明显低于对照组和空载体组(20.13±7.00和19.70±6.15,P<0.05)。结论:转染NK4基因可显著抑制裸鼠胰腺癌移植瘤的生长,其作用机制可能是通过抑制肿瘤新生血管的形成,促进肿瘤细胞凋亡。

转染NK4基因对人胰腺癌SW1990细胞生物学特性的影响

背景与目的:肝细胞生长因子(hepatocytegrowthfactor,HGF)通过其受体c-Met的激活,在调节肿瘤侵袭和转移和血管形成中具有重要作用。NK4不仅是HGF的拮抗剂,也是血管形成的抑制剂,阻断HGF/c-Met途径和肿瘤血管的形成可成为抗肿瘤治疗的策略之一。为此,我们构建NK4基因真核细胞表达载体并进行转染研究,探讨NK4基因在胰腺癌细胞中的表达及其对胰腺癌细胞生物学特性的影响。方法:对重组pcDNA3/hNK4质粒进行酶切,将NK4基因克隆到真核表达载体pRC/CMV2,应用脂质体将重组pRC/CMV2-hNK4质粒转入胰腺癌SW1990细胞中,采用逆转录聚合酶链反应(RT-PCR)及免疫印迹(Westernblot)分别检测转染的肿瘤细胞中NK4mRNA和蛋白的表达并筛选出高表达的细胞克隆。采用Transwall小室和Matrigel侵袭小室测定转染NK4基因对胰腺癌细胞运动和侵袭的影响。结果:转导NK4基因的SW1990细胞可表达并分泌NK4,RT-PCR扩增出预期的453bp片段,Westernblot显示有50000的NK4蛋白,转染NK4基因对胰腺癌细胞SW1990的生长无抑制作用(P>0.05),但可显著抑制HGF或成纤维细胞所诱导胰腺癌细胞的运动和侵袭(P<0.01),而转染空载体则无此作用(P>0.05)。结论:转染NK4基因可抑制胰腺癌细胞的运动和侵袭,在胰腺癌抗转移治疗中可能具有重要作用。

大豆中Glycinol的分离与鉴定

Glycinol是大豆中主要的植物抗毒素和植物雌激素glyceollins的直接前体,是研究大豆中异戊烯转移酶必需的重要底物。该研究以新鲜大豆为原料,采用硝酸银诱导大豆子叶使glycinol在其中累积,再用甲醇提取,经制备高效液相色谱分离获得单一成分;经液相色谱/四极杆-飞行时间串联质谱及紫外光谱鉴定,并与标准品对照,确定所获得的单一成分为glycinol。采用该分离方法可从390g鲜重的大豆中分离得到100mg高纯度的glycinol。该研究提供了一种分离纯化和较大量制备glycinol的方法。

含patatin-GUS嵌合基因的质粒的构建及其在马铃薯中的表达

本试验构建了一个含有马铃薯蛋白(patatin)基因的5′端调控序列和细菌葡糖苷酸酶(β-glu-curonidase GUS EC 3.2.1.31)的编码序列及胭脂碱合成酶3′端序列所组成的嵌合基因的重组质粒。利用三亲交配法将其导入经改造的根癌农杆菌菌株Agrobacterium tumefaciensLBA 4404,然后利用农杆菌介导,将该嵌合基因转入马铃薯栽培品种Desiree。从卡那霉素抗性筛选得到的再生的转基因植株,经扩大培养后,对其叶、茎、根和块茎等组织的抽提液的GUS酶活力分析表明,嵌合基因在转基因植株中得到了明显的表达。光照条件下培养和完全黑暗中培养的转基因植株的幼苗的GUS酶活力分析还表明,嵌合基因在转基因植株中的表达可能是受到光的诱导的。

蚯蚓组织中硫氧还蛋白还原酶抑制剂初探

目的研究蚯蚓组织成份对硫氧还蛋白系统的影响。方法乙酸盐缓冲液抽提蚯蚓组织成份 ,不同处理的蚯蚓提取液与硫氧还蛋白系统反应。在 340nm波长处 ,用NADPH作为被监测的反应底物来动态监测硫氧还蛋白还原酶 (TrxR)的催化活性。结果蚯蚓提取液与TrxR单独作用后 ,监测底物NADPH的吸光值几乎不发生变化。蚯蚓提取液的溶剂及煮沸后的上清液不具有以上作用。结论蚯蚓组织中含有能够抑制TrxR活性的物质 ,该物质的化学本质为蛋白质。