秦川牛宰后成熟过程中线粒体Tu翻译延长因子对牛肉持水性的影响

探究秦川牛宰后成熟过程中线粒体Tu翻译延长因子(mitochondrial Tu translation elongation factor,TUFM)表达对肉的持水性影响。以秦川牛背最长肌为研究对象,测定4℃不同成熟时间下的pH值、贮藏损失、离心损失、蒸煮损失、水分分布、肌原纤维蛋白等指标变化情况,测定不同成熟时间(0、96、192 h)下TUFM表达量及其含量、Beclin1蛋白表达量。结果显示:在秦川牛宰后成熟期间,肌原纤维蛋白发生降解,TUFM的表达量与Beclin1蛋白表达量和牛肉的持水性存在密切关系,其中蛋白质组学测定的TUFM表达量变化与TUFM含量变化趋势一致,Beclin1蛋白表达量、贮藏损失、离心损失、蒸煮损失整体均呈先上升后下降趋势,pH值呈先下降后上升趋势;Pearson相关性分析表明,牛背最长肌中TUFM表达量与低场核磁共振峰面积比P_(2b)、Beclin1蛋白表达量呈极显著正相关(P<0.01),与贮藏损失、离心损失、蒸煮损失呈显著正相关(P<0.05),与P_(21)呈极显著负相关(P<0.01),与P_(22)呈显著负相关(P<0.05),与pH值无显著相关性(P>0.05)。通过蛋白质组学鉴定出23种与TUFM相关的差异蛋白,通过基因本体论、京都基因与基因组百科全书通路分析发现,差异蛋白可通过多种途径参与能量代谢,进而介导细胞自噬;对差异蛋白和持水性指标进行Pearson相关性分析发现,有5种差异蛋白(ATP5F1D、EEF1A2、GSPT1、NDUFB5、SUCLG1)与持水性指标具有显著相关性(P<0.05、P<0.01)。分析可知,包括TUFM在内,共6种蛋白主要通过能量代谢和氧转运等途径正向或负向影响细胞自噬,从而影响肉的持水性。

Patch testing and cross sensitivity study of adverse cutaneous drug reactions due to anticonvulsants: A preliminary report

AIM To evaluate the utility of patch test and cross-sensitivity patterns in patients with adverse cutaneous drug reactions(ACDR) from common anticonvulsants. METHODS Twenty-four(M:F = 13:11) patients aged 18-75 years with ACDR from anticonvulsants were patch tested 3-27 mo after complete recovery using carbamazepine, phenytoin, phenobarbitone, lamotrigine, and sodium valproate in 10%, 20% and 30% conc. in pet. after informed consent. Positive reactions persisting on D3 and D4 were considered significant. RESULTS Clinical patterns were exanthematous drug rash with or without systemic involvement(DRESS) in 18(75%), Stevens-Johnsons syndrome/toxic epidermal necrolysis(SJS/TEN) overlap and TEN in 2(8.3%) patients each, SJS and lichenoid drug eruption in 1(4.2%) patient each, respectively. The implicated drugs were phenytoin in 14(58.3%), carbamazepine in 9(37.5%), phenobarbitone in 2(8.3%), and lamotrigine in 1(4.7%) patients,respectively. Twelve(50%) patients elicited positive reactions to implicated drugs; carbamazepine in 6(50%), phenytoin alone in 4(33.3%), phenobarbitone alone in 1(8.3%), and both phenytoin and phenobarbitone in 1(8.33%) patients, respectively. Cross-reactions occurred in 11(92%) patients. Six patients with carbamazepine positive patch test reaction showed cross sensitivity with phenobarbitone, sodium valproate and/or lamotrigine. Three(75%) patients among positive phenytoin patch test reactions had cross reactions with phenobarbitone, lamotrigine, and/or valproate. CONCLUSION Carbamazepine remains the commonest anticonvulsant causing ACDRs and cross-reactions with other anticonvulsants are possible. Drug patch testing appears useful in DRESS for drug imputability and cross-reactions established clinically.

Anticonvulsant potential of rosuvastatin in combination with carbamazepine and valproate in animal models of epilepsy

BACKGROUND Epilepsy impacts millions of people,with many not responding to existing treatments.Some evidence links neuroinflammatory processes to epilepsy.Statins exhibit anti-inflammatory and neuroprotective properties,potentially offering antiepileptic effects.AIM To evaluate the anticonvulsant effects of rosuvastatin in animal models of epilepsy.METHODS Ninety-six albino mice were divided into 16 groups.In the maximal electroshock seizure(MES)model,eight groups received intraperitoneal vehicle,carbama-zepine,rosuvastatin,or a combination.Outcomes measured included seizure protection[tonic hind limb extension(THLE)],duration of THLE,seizure duration,and mortality.In the pentylenetetrazol(PTZ)model,eight groups were pretreated with vehicle,valproate,rosuvastatin,or a combination,with outcomes measured as seizure latency,seizure duration,and mortality.RESULTS In the MES model,rosuvastatin exhibited protection against THLE in a small percentage of mice.Rosuvastatin shortens the duration of THLE in a dose-dependent manner.However,none of these were statistically significant com-pared to the control group.The combination of rosuvastatin 10 mg/kg with carbamazepine 4 mg/kg resulted in a significant reduction in seizure duration compared to the control group,better than carbamazepine alone at 4 mg/kg and 6 mg/kg.In the PTZ model,rosuvastatin alone showed no significant effects on latency,duration of seizure,or mortality.However,rosuvastatin 10 mg/kg combined with valproate 100 mg/kg significantly delayed the onset of seizures,seizure duration and mortality percentage,better than valproate alone at 100 mg/kg.CONCLUSION Rosuvastatin enhanced the anticonvulsant effects of carbamazepine and valproate.Further studies are required to explore the antiepileptic potential of rosuvastatin at various doses,durations,dosage forms,routes and models.

A Study of the Relationship Between the Chemical Structure and the Anticonvulsant Activity of the 3-Pyrazolidinones

The Hansch approach was used in the quantitative structure anticonvulsant activity studies of the previously synthesized 1-substituted-and 1.5-disubstituted-3-pyrazo- lidinones.Correlation analysis predicted that 1.5-disubstituted-3-pyrazoljdinones in which 1- substituent is n-propyl but not benzyl.with the total hydrophobic constant of 1-and 5-sub- stituents(∑л)equals to 4.5(optimum value)will have the most potent activity.On the basis of this analysis eleven 5-substituted and I-n-butyl-5-substituted-3-pyrazolidinones were syn- thesized.Pharmacological tests indicated that the prediction of the Hansch analysis of the 3- pyrazolidinones is correct.The Hansch analysis,by including these 11 compounds,gives an almost identical correlation with that previously obtained.

Studies on the Synthesis, Anticonvulsant Activity, and the Structure-Activity Relationships of Phenyl Pyridazinones and their GABA Derivatives

In searching for effective anticonvulsant agents,fourteen 6-aryl-4.5-di- hydro-3(2H)pyridazinones.fifteen 6-aryl-3(2H)pyridazinones,and seventeen 3-GABA derivatives of 6-aryIpyridazines have been synthesized,and evaluated in mice for the ability to antagonize maximal electroshock seizure(MES).The ED_(50) values showed that 6-(2′,4′- dichlorophenyt)-3(2H)pyridazinone was the most potent anticonvulsant among these corn- pounds(ED_(50)=10.15 mg/kg).The structure-activity relationships of the aryl pyridazinones were studied.The result showed that:(1)the higher the value of the hydrophobic parameter л of the substituent on the phenyl ring.the more potent the anticonvulsant activity of the corn- pound.and(2)only the compounds with an electron withdrawing substituent on the phenyl ring exhibited appreciable anticonvulsant activity.

工程教育中心何以推动科教融合——荷兰4TU工程教育中心的探索性单案例研究

工程教育中心作为建立在大学或研究机构中的跨学科交叉合作平台,是连接科学研究与教育实践的纽带,在高质量工程人才培养中发挥着重要作用。荷兰4TU工程教育中心利用4所顶尖理工大学在工程学科和教育领域的独特优势,积极与研发单位、教育单位、企业部门合作,通过将前沿科学研究彻底融入工程课程设计、教学模式等多个方面,形成了独具一格的科教融合工程人才培养模式。文章从战略目标、组织架构、运行机制、质量保障4个维度详实分析了4TU工程教育中心推动科教融合的内在机制,总结归纳其在主题项目设置、教育共同体形成、课程体系迭代、创新网络构建、内外部质量保障等方面的核心特征,期望对我国科教融合的工程教育改革与建设有所启示。

A novel series of 2,5-disubstituted 1,3,4-thiadiazoles as potential anticonvulsant agent

In pursuit for better antiepileptic drug and the importance of semicarbazones and 2,5-disubstituted 1,3,4-thiadiazoles as anticonvulsant pharmacophore, a series of novel N-（{5-[（6-methyl-l-benzofuran-3-yl）methyl]-l,3,4-thiadiazol-2-yl}carba- mothioyl）-2/3/4-substitutedbenzamide were designed, synthesized and evaluated for their anticonvulsant activity. The findings of the present studies confirmed that the pharmacophore model with four binding sites is crucial for anticonvulsant activity. Structure-activity relationships among synthesized compounds were also established.

沉默TUFM通过AMPK/mTOR信号通路调控线粒体自噬对肺源性心脏病模型大鼠肺动脉高压的影响

目的探讨线粒体翻译延伸因子Tu(TUFM)通过线粒体自噬促进肺动脉高压(PAH)血管重塑的作用机制。方法2022年1月—2023年6月于辽宁省人民医院中心实验室进行实验。将36只健康雄性Sprague-Dawley大鼠随机分为空白对照(Ctrl)组、模型(PAH)组、TUFM过表达(OE)组、OE阴性对照(OE-NC)组、短发夹RNA(Sh)敲除TUFM(Sh)组和Sh-NC阴性对照(Sh-NC)组,每组6只。除Ctrl组外,其余大鼠均一次性腹腔注射1%野百合碱(60 mg/kg)诱导心源性肺水肿PAH大鼠模型;大鼠肺动脉平滑肌细胞(PASMC)在低氧(3%O 2)条件下培养24 h模拟体内肺动脉高压微环境,分为常氧(Norm)组、低氧(Hyp)组、小干扰RNA(SiRNA)-1组、SiRNA-2组、Si-NC组、OE-NC组和OE组。右心导管插管和脉冲多普勒超声检测大鼠肺血流动力学;苏木素-伊红染色检测肺小动脉病理结构;免疫荧光共染检测TUFM组织定位;细胞计数法检测细胞增殖;透射电镜观察线粒体结构和自噬小体;蛋白免疫印迹检测TUFM、自噬、凋亡和磷酸腺苷活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)通路相关蛋白表达。结果与Ctrl组比较,PAH组大鼠TUFM蛋白表达升高,且主要与PASMC标志物α平滑肌肌动蛋白(α-SMA)在肺小动脉内膜存在共定位,而与内皮细胞标志物CD31无共定位,肺动脉收缩压(PASP)升高,肺动脉血流加速时间(PAAT)缩短,远端肺小动脉管壁呈向心性增厚,管腔狭窄几乎堵塞,TUFM、苄氯素1重组蛋白(BECN1)、人微管相关蛋白轻链3(LC3)II/I和B淋巴细胞瘤2(Bcl2)蛋白表达升高,P62、Bcl2相关X蛋白(Bax)和凋亡酶激活因子(Apaf)蛋白表达降低(P<0.05);与PAH组比较,OE组PASP升高,PAAT缩短,肺小动脉管壁厚度升高,肺动脉TUFM、BECN1、LC3II/I和Bcl2表达升高,P62、Bax和Apaf表达降低(P<0.05);与PAH组比较,Sh组PASP降低,PAAT延长,肺小动脉管壁厚度和管腔狭窄度有所改善,TUFM、BECN1、LC3II/I和Bcl2表达降低,P62、Bax和Apaf表达升高(P<0.05)。与Norm组比较,Hyp组PASMC细胞TUFM蛋白表达升高;与Si-NC组细胞相比,SiRNA-1和SiRNA-2组P62、Bax蛋白表达升高,BECN1、LC3II/I、Bcl2、TUFM表达降低,线粒体结构完整,PASMC细胞增殖活性降低,细胞p-AMPK表达降低,p-mTOR表达升高(P<0.05);与OE-NC组比较,OE组细胞P62和Bax蛋白表达降低,BECN1、LC3II/I、Bcl2和TUFM表达升高,部分线粒体损伤崩解,嵴断裂消失,PASMC细胞增殖活性明显升高,细胞p-AMPK表达升高,p-mTOR表达降低(P<0.05)。结论沉默TUFM可通过激活AMPK/mTOR信号通路促进线粒体自噬加速PAH肺动脉平滑肌细胞凋亡。

秦川牛宰后成熟过程中线粒体翻译延长因子Tu与能量代谢的关联性分析

目的:线粒体翻译延长因子Tu(mitochondrial Tu translation elongation factor,TUFM)已被证明参与秦川牛宰后成熟过程中的细胞自噬活动,实验探究该过程中TUFM表达与能量代谢变化的关系。方法:以秦川牛背最长肌为研究对象,测定4℃不同成熟时间(0、2、12、24、48、96、144、192 h)TUFM表达量及含量、葡萄糖(glucose,GLU)、乳酸(lactic acid,LA)、腺苷三磷酸(adenosine triphosphate,ATP)、二磷酸腺苷(adenosine diphosphate,ADP)、单磷酸腺苷(adenosine monophosphate,AMP)6种物质含量以及乳酸脱氢酶(lactate dehydrogenase,LDH)、琥珀酸脱氢酶(succinate dehydrogenase,SDH)、磷酸丙糖异构酶(triose phosphate isomerase,TPI)、苹果酸脱氢酶(malate dehydrogenase,MDH)、细胞色素c氧化酶(cytochrome c oxidase,COX)5种酶活性的变化情况。结果:在秦川牛宰后成熟期间,GLU、ATP、ADP、AMP含量和LDH、SDH、TPI活性均呈下降趋势,TUFM表达量、MDH活性及LA和TUFM含量均呈先上升后下降趋势,COX活性呈先下降后上升再下降趋势。能量代谢各指标和TUFM蛋白主要在宰后初期发挥作用,对宰后初期及宰后中后期分别进行Pearson相关性分析,结果表明,在宰后初期牛背最长肌中TUFM表达量与MDH、LA呈极显著正相关(P<0.01),与ATP、ADP、AMP、LDH、SDH、TPI、COX、GLU呈极显著负相关(P<0.01)。在宰后中后期,TUFM表达与所有指标(GLU、LA、ATP、ADP、AMP、LDH、MDH、SDH、TPI、COX)均呈极显著正相关(P<0.01)。结论:在宰后初期TUFM表达量逐渐增加,可为肌肉细胞提供能量从而用于能量代谢途径,该过程可能是TUFM正向参与细胞自噬所致。在宰后中后期能量短缺时,TUFM可能抑制细胞自噬,优先将能量用于除细胞自噬外的其他重要途径(如能量代谢途径)以维持细胞稳态。综上,在宰后成熟过程中TUFM具有双重作用,可调节细胞自噬为能量代途径提供能量,有助于维持宰后能量代谢持续的时间。

绵羊肺炎支原体EF-Tu蛋白的原核表达及多克隆抗体制备

[目的]克隆绵羊肺炎支原体(Mycoplasma ovipneumoniae,Mo)EF-Tu基因,原核表达获得EF-Tu蛋白,制备抗EF-Tu蛋白的兔源多克隆抗体,为研究肺炎支原体EF-Tu蛋白的结构和功能奠定基础。[方法]采用重叠延伸PCR方法将pET-28a-EF-Tu质粒中EF-Tu基因中间的TGA密码子突变为TGG,并对测序结果与其他支原体参考株进行相似性比对和遗传进化分析,利用在线软件对其推测的蛋白序列进行生物信息学分析。将突变后的重组质粒转化大肠杆菌BL21(DE3)感受态细胞进行原核表达,经SDS-PAGE和Western blotting鉴定,利用镍柱亲和层析法纯化,以纯化的EF-Tu融合蛋白免疫家兔制备多克隆抗体,采用间接ELISA和Western blotting检测多克隆抗体效价及免疫反应性。[结果]试验成功突变了EF-Tu基因中TGA位点,并构建了融合表达His标签pET-28a-EF-Tu′原核表达载体。生物信息学分析表明,克隆的EF-Tu基因与绵羊肺炎支原体MoGH3-3菌株相似性最高,亲缘关系最近;编码387个氨基酸,无N-糖基化位点和跨膜区域,存在10个丝氨酸、20个苏氨酸、4个酪氨酸磷酸化位点,二级结构由无规则卷曲(35.14%)、α-螺旋(26.87%)、延伸链(26.87%)及β-转角(11.11%)构成。SDS-PAGE和Western blotting结果显示,目的蛋白大小约为43 ku,蛋白纯化浓度为0.615 g/L。ELISA和Western blotting结果显示,制备的多克隆抗体效价可达1∶128 000,能够特异性识别EF-Tu融合蛋白,具有良好的免疫反应性。[结论]本研究成功突变了EF-Tu基因的TGA密码子,原核表达并纯化获得EF-Tu融合蛋白,制备其多克隆抗体效价为1∶128 000,为后续研究肺炎支原体EF-Tu蛋白结构和生物学功能及其疫苗研发提供了试验基础。

Studies on the Relationship Between the Structure and Anticonvulsant Activity of Cinnamamides and Their Analogs

On the basis of systematic modification of the structure of componds of antiepilepsirine type, more than 200 cinnamamides were synthesized and tested by animal assay (maximal electroshock seizure, MES). Pharmacological evaluation showed that the configuration and the substituents on the phenyl ring and the nitrogen of amides, and substituents on the double bond displayed an important effect on the anticonvulsant activity.For studying the effect of the modification of structure to anticonvulsant activity, Hansch approach was employed to study the QSAR among 38 cinnamamides, and Hopfinger' s MSA was employed to study the MSA-QSAR among 26 cinnamamides.

异氟烷麻醉对小鼠自发肌电及TUS/TMAS诱发肌电的影响

经颅超声刺激(TUS)和经颅磁声耦合刺激(TMAS)调控运动皮层效果明显,但受限于清醒状态动物难以束缚,已有研究大多在麻醉状态下进行,对麻醉减弱调控效果的分析集中于中枢神经系统。本研究记录了异氟烷麻醉下24只小鼠的肢体自发肌电和TUS/TMAS诱发肌电,定量分析了麻醉对自发肌电和诱发肌电发放率、潜伏期、时长和幅值的影响。结果显示,随着异氟烷输出浓度从0.40%增加至0.75%,每周期内小鼠自发肌电频次减少约50%,肌电发放时长变短,呈抑制状态;TUS/TMAS诱发肌电的成功率分别降低约50%和70%、潜伏期均延长约0.1 s、时长分别缩短约0.3和0.5 s,表明TUS/TMAS对运动皮层的调控效果随麻醉程度的加深而减弱。肢体自发和诱发肌电在发放率和时长上存在关联性特征,提示麻醉状态下小鼠自发肌电抑制状态可能是刺激效果减弱的影响因素之一。

Anticonvulsant, Sedative and Antidepressant Effects of Aqueous Extract of Costus afer Stems in Mice

Epilepsy is a disorder in the nervous system which often causes a loss of consciousness. Traditional treatments are quiet a component of health care system in various populations in spite of the fact that well-established options are available. Most plants are used to treat epilepsy or those which have been verified for anticonvulsant activity were reported. Then, Costus afer is a plant of the Congolese flora used in traditional medicine for its many virtues. Therefore, the anticonvulsant activity of Costus afer was assessed with the strychnine convulsion induction test. Two tests were used for sedative activity such as the barbiturate sleep induction test and motor activity and finally the forced swimming test was also used to assess antidepressant activity. The results showed that the aqueous extract of Costus afer stems had no effects on strychnine-induced seizures at doses of 250 mg/kg and 500 mg/kg compared to the control group. However, the extract of Costus afer stems caused a very significant decrease in motricity at a dose of 500 mg/kg, showing a decrease in the onset time and a very significant increase in sleep duration like the reference molecule such as Diazepam. The aqueous extract of Costus afer stems also caused a decrease in immobility time in mice at a dose of 500 mg/kg.

Synthesis and evaluation of 4-substituted semicarbazones of levulinic acid for anticonvulsant activity

Objective: A series of 4-aryl substituted semicarbazones of levulinic acid (4-oxo pentanoic acid) was designed and synthesized to meet the structural requirements essential for anticonvulsant activity. Methods: All the compounds were evaluated for anticonvulsant activity. Anticonvulsant activity was determined after intraperitoneal (i.p.) administration to mice by maximal electroshock (MES) and subcutaneous metrazol (ScMet) induced seizure methods and minimal motor impairment was determined by rotorod test. Results: A majority of the compounds exhibited significant anticonvulsant activity after intraperitoneal administration. In the present study 4-(4'-fluoro phenyl) levulinic acid semicarbazone emerged as the most active molecule, showing broad spectrum of activity with low neurotoxicity. Unsubstituted levulinic acid semicarbazone was found to be inactive in all the screens. Conclusion: The results obtained validate the hypothesis that presence of an aryl group near the semicarbazone moiety is essential for anticonvulsant activity. The results also indicate that the hydrophilic-hydrophobic site can accommodate hydrophilic groups.

Molecular Docking Studies on Anticonvulsant Enaminones Inhibiting Voltage-Gated Sodium Channels

Epilepsy is described as the most common chronic brain disorder. A typical symptom of epilepsy results in uncontrolled convulsions caused by temporary excessive neuronal discharges. Although several new anticon-vulsants have been introduced, some types of seizures have still not been adequately controlled with these new and current therapies. There is an urgent need to develop new anticonvulsant drugs to control the many different types of seizures. Many studies have shown that the epilepsies involve more than one mechanism and therefore may be responsible for the various types of observed seizures. Recently reported studies have shown that a group of newly synthesized 6 Hz active anticonvulsant fluorinated N-benzamide enaminones exhibited selective inhibitions of voltage-gated sodium (Nav) channels. Nav channels are responsible for the initial inward currents during the depolarization phases of the action potential in excitable cells. The activation and opening of Nav channels result in the initial phases of action potentials. We hypothesize that there is an essential pharmacophore model for the interactions between these enaminones and the active sites of Nav channels. The research reported here is focused on molecular docking studies of the interactions that occur between the fluorinated N-benzamide enaminones and the Nav channels. These studies may open an avenue for designing anticonvulsant drugs by inhibiting Nav channels.

Microwave-assisted synthesis,anticonvulsant activity and quantum mechanical modelling of N-(4-bromo-3-methylphenyl) semicarbazones

Objective: To study the effect of halo substitution on disubstituted aryl semicarbazones on the anticonvulsant potential and model the activity based on quantum mechanics. Methods: A series of twenty-six compounds of N4-(4-bromo-3-methylphenyl) semicarbazones were synthesized and evaluated for the anticonvulsant activity in the maximal electroshock seizure (MES) and subcutaneous pentylenetetrazole (scPTZ) seizure threshold tests. Some potential compounds were also tested in the subcutaneous strychnine (scSTY) and subcutaneous picrotoxin (scPIC) seizure threshold tests. The synthesized compounds were tested for behavioral impairment and CNS (central nervous system) depression in mice. Quantum mechanical modelling was carried out on these compounds to gain understanding on the structural features essential for activity. Results: Some compounds possessed broad spectrum anticonvulsant activity as indicated by their effect in pentylenetetrazole, strychnine, picrotoxin and maximal electro- shock seizures models in resemblance to other aryl semicarbazone derivatives reported earlier. The higher the difference in HOMO (highest occupied molecular orbital) and LUMO (lowest unoccupied molecular orbital) energy levels was, the greater was the activity profile. Conclusion: The pharmacophoric requirements for compounds to exhibit anticonvulsant activity that includes one aryl unit in proximity to a hydrogen donor-acceptor domain and an electron donor have been justified with the molecular orbital surface analysis of the synthesized compounds.

An Investigation of Moxibustion Treatment for Abscesses in the Song Dynasty:Focusing on the“Jiu Ai Tu”

Jiu Ai Tu(The Moxa Treatment)from the Song dynasty is the earliest surviving painting that focuses on the subject of acupuncture and moxibustion.This paper takes the medical activities depicted in the artwork as its research object and systematically analyzes the external treatment methods for abscesses during the Song dynasty reflected in Jiu Ai Tu.By examining the understanding of abscesses during that period,the paper explores the level of development in external medicine techniques.By analyzing the medical awareness and behaviors of patients when facing such severe illnesses,it aims to explore the societal cognition and experiences regarding health and disease.The paper attempts to present the folk medical ecology of the Song dynasty represented by Jiu Ai Tu.

Investigation of Some Possible Mechanisms Involved in the Anticonvulsant Activity of Tulbaghia violacea Harv

Even though Tulbaghia violacea has been used to treat and manage epilepsy in South Africa by traditional medicine practitioners, no evidence in any literature has shown any scientific scrutiny of the effectiveness of the plant species in therapy. This study was intended, therefore, to investigate the anticonvulsant effect of the leaf methanol extract of Tulbaghia violacea by studying its effect against tonic convulsion induced by either PTZ （pentylenetetrazole）, bicuculline, picrotoxin, strychnine or NMDLA （N-methyl-DL-aspartic acid） in mice. Qualitative phytochemical analysis, acute toxicity and HPLC studies were also carried out on the plant species. Leaf methanol extract of Tulbaghia violacea, phenobarbitone, diazepam or muscimol significantly antagonised PTZ, bicuculline or picrotoxin-induced convulsion. Combined treatment of sub-effective doses of T. violacea and muscimol significantly antagonised tonic convulsion induced by PTZ. T. violacea or phenobarbitone significantly antagonised strychnine-induced tonic convulsion. T. violacea or LY233053 significantly antagonised NMDLA-elicited tonic convulsion. Phenytoin or DMSO （dimethylsulfoxide） did not significantly affect the tonic convulsion produced by PTZ, bicuculline, picrotoxin, strychnine or NMDLA. The phytochemical qualitative analysis of the plant species showed the presence of alkaloids, saponins, reducing sugars, flavonoids, cardiac glycosides, triterpene steroids, quinones and tannins. The LD50 value obtained following oral administration of the plant extract was over 4000 mg/kg. The data in the present study indicate that the leaf methanol extract of T. violacea has anticonvulsant activity which is probably underpinned by GABAergic, glutaminergic and glycinergic mechanisms.

The water-soluble TF3 component from Eupolyphaga sinensis Walker promotes tibial fracture healing in rats by promoting osteoblast proliferation and angiogenesis

Objective:To determine the active components of Eupolyphaga sinensis Walker(Tu Bie Chong)and explore the mechanisms underlying its fracture-healing ability.Methods: A modified Einhorn method was used to develop a rat tibial fracture model.Progression of bone healing was assessed using radiological methods.Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site.Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration.The Transwell assay was used to explore the invasion capacity of the cells.Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells(HUVECs).qRT-PCR was used to evaluate the changes in gene transcription levels.Results: Tu Bie Chong fraction 3(TF3)significantly shortened the fracture healing time in model rats.X-ray results showed that on day 14,fracture healing in the TF3 treatment group was significantly better than that in the control group(P=.0086).Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group.In vitro,TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs(all P<.01).Transwell assays showed that TF3 promoted the migration of HUVECs,but inhibited the migration of MC3T3-E1 cells.Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules.Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA,SPOCD1,NGF,and NGFR in HUVECs.In MC3T3-E1 cells,the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment.Conclusion: TF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.

黄瓜(Cucumis sativus L.)果瘤基因Tu的精细定位

黄瓜果实果瘤性状是非常重要的农艺性状,以黄瓜(Cucumis sativus L.)华北有瘤品系S52(P1)和欧洲无瘤品系S06(P2)为亲本,构建826株F2(S06×S52)群体为试验材料,对黄瓜果瘤基因Tu进行精细定位研究。田间调查和遗传分析发现F2群体的果实表现型为有果瘤与无果瘤,卡方值(χ2=0.583<3.84)检验表明其分离比符合3∶1,表明黄瓜果瘤性状是单基因控制的显性性状。早期研究中Tu基因被初步位于第5染色体的标记SSR16203和C_SC933之间,本研究利用公布的黄瓜基因组序列在这两标记之间设计了100对SSR引物,通过分析在两亲本的多态性,最后筛选得到4对SSR多态性标记。然后利用SSR16203和C_SC933标记和这4个多态性引物对826株F2群体进行进一步交换单株分析,最后将Tu基因定位于两翼标记SSR7和SSR18之间,剩余的交换单株分别为11株、16株,物理距离为263kb。经过FGENESH基因预测软件(http:∥www.softberry.com)分析,该区间仅含有候选基因31个。本研究为进一步果瘤基因Tu的最后克隆提供良好的研究基础。