Exploring the anti-aging effects of chlorogenic acid and the underlying mechanisms based on a Caenorhabditis elegans model

Objective:To explore the anti-aging effects of chlorogenic acid(CGA)and the underlying mechanisms based on a Caenorhabditis elegans(C.elegans)model.Methods:The anti-agingactivityofCGAwasstudied basedon thebodylength,exercisebehavior,lipofuscin content,antioxidative stress ability,swallowing frequency,body-bending frequency,and head-swinging ability of C.elegans.Through DAF-16 nuclear translocation and SOD-3-GFP fluorescence experiments,the effects of CGA on ROS levels,antioxidant enzyme activities,MDA content,mutant-strain lifespan,and anti-aging molecular signaling pathways were explored,as well as the underlying mechanisms.Results:CGA improved multiple indices of the nematode:body length was increased(all P<0.001),head-swing frequency and body-bending frequency were increased(all P<0.05),nematode longevity was prolonged(P=0.0021),lipofuscin deposition in nematodes was slowed down(all P<0.001),the chemotaxis index was improved(P=0.0012),ROS levels were reduced(all P<.001),and SOD activity and MDA content were reduced(SOD:P=0.0017 between the low-concentration group and the control group,P<.001 between the high-concentration and medium-concentration groups and the control group;MDA:P=0.0135 between the low-concentration group and the control group,and P<0.001 between the high-concentration and medium-concentration groups and the control group).In addition,CGA also activated the DAF-16 transcription factor,promoted DAF-16 nuclear translocation under oxidative stress conditions(both P<0.001 between the high-concentration and medium-concentration groups and the control group),and increased SOD-3 gene expression in nematodes(all P<0.001).Conclusion:CGA plays an anti-aging role in C.elegans.The underlying mechanisms include activation of the insulin/IGF-1 signaling pathway and enhancement of DAF-16 activity.This study lays a foundation for further research into the anti-aging effects of CGA.

Antiviral Activity of the Effective Monomers from Folium Isatidis Against Influenza Virus in Vivo

In order to evaluate the anti-influenza virus activity of the effective monomer from Folium Isatidis (FI) in vivo,we established mice model with viral pneumonia and divided them into 3 different dose groups,then observed their lung indexes,pulmonary pathological changes,pulmonary virus hemagglitination titers,living time and death rates.The results showed that the monomer could reduce the pulmonary index from 2.64 to 1.93,1.63 and 1.40 (P<0.01) and decrease the hemagglitination titer from 1.15 to 0.84,0.70 and 0.59 (P<0.01).In addition,different groups of FI could significantly lessen the mortality rate from 100% to 30%,25% and 15%,and prolong the living time from 5.1d to 6.5d,8.4d and 8.9d respectively(P<0.01).The high dose (75 mg/kg/d) has the similar effect with 100 mg/kg/d dose of virazole(P>0.05),and more effective than 200 mg/kg/d dose of antiviral liquor (P<0.05).

<i>In Vivo</i>Antioxidant Activity of Fucoxanthin on Obese/Diabetes KK-<i>A^y</i>Mice

Dietary intake of 0.1% fucoxanthin significantly reduced lipid hydroperoxide levels of liver and abdominal white adipose tissue (WAT) of obese/diabetes KK-Ay mice. The fucoxanthin supplementation also significantly reduced blood glucose level and hepatic lipid contents of the mice. Oxidative stress is known to be induced in hyperglycemia and high fat conditions. Therefore, in vivo antioxidant activity of fucoxanthin found in the present study could be attributed to its anti-diabetic effect and its decreasing effect on hepatic lipids. On the other hand, little effect of fucoxanthin on lipid hydroperoxide levels was found in normal ICR mice. Although the content of fucoxanthin metabolites in the abdominal WAT of KK-Ay mice was about 50 times higher that in the liver, there was little difference in its In Vivo antioxidant activity between in the liver and in the abdominal WAT. These results suggest that well-known ability of fucoxanthin to scavenge active oxygen species and/or free radicals would not be a main reason to explain its In Vivo antioxidant activity.

Conversion of a normal maize hybrid into a waxy version using in vivo CRISPR/Cas9 targeted mutation activity

Waxy maize is a specialty maize that produces mainly amylopectin starch with special food or industrial values. The objective of this study was to overcome the limitations of wx mutant allele acquisition and breeding efficiency by conversion of parental lines from normal to waxy maize. The intended mutation activity was achieved by in vivo CRISPR/Cas9 machinery involving desired-target mutation of the Wx locus in the ZC01 background,abbreviated as ZC01-DTM^(wx). Triple selection was applied to segregants to obtain high genome background recovery with transgene-free wx mutations. The targeted mutation was identified, yielding six types of mutations among progeny crossed with ZC01-DTM^(wx).The amylopectin contents of the endosperm starch in mutant lines and hybrids averaged94.9%, while those of the wild-type controls were significantly(P < 0.01) lower, with an average of 76.9%. Double selection in transgene-free lines was applied using the Bar strip test and Cas9 PCR screening. The genome background recovery ratios of the lines were determined using genome-wide SNP data. That of lines used as male parents was as high as98.19% and that of lines used as female parents was as high as 86.78%. Conversion hybrids and both parental lines showed agronomic performance similar to that of their wild-type counterparts. This study provides a practical example of the efficient extension of CRISPR/Cas9 targeted mutation to industrial hybrids for transformation of a recalcitrant species.

Investigation of the Antimicrobial Activity and <i>in Vivo </i>Cytotoxicity of <i>Diospyros malabarica </i>(Desr.) Kostel. Fruit Extracts

Mankind is facing an unprecedented threat of existence due to the antibiotic resistance developed by bacteria. The unripe fruits of Diospyros malabarica (Desr.) Kostel. (family: Ebenaceae) can be considered as one of the natural sources to tackle this issue. The present study is designed to assess the antimicrobial activity of D. malabarica seed and flesh ex-tracts. Herein, D. malabarica extracts were prepared using polar solvents (i.e., water and 70% ethanol) and their antimicrobial activity as well as in vivo toxicity was investigated. Their antibacterial activity was investigated against gram positive (Bacillus subtilis) and gram negative (Escherichia coli DH5α, and Salmonella typhi) bacteria at different time points. All the extracts showed the highest antibacterial activity after 2 hours of incubation. The aqueous seed extract showed the maximum zone of inhibition (i.e., ~13 mm) against Bacillus subtilis with a minimum inhibitory concentration (MIC) value of 2 μg/μl. The an-tibacterial propensity was also confirmed through trypan blue dye exclusion assay, CellToxTM Green assay, and lipid peroxidation (LPO) assay. On the other hand, the etha-nolic seed extract demonstrated higher antifungal activity through inhibition of mycelial growth. All the extracts showed excellent hemocompatibility against both human and rat red blood cells (RBCs). They also did not show any toxicity to rat liver and kidneys. Taken together, this study demonstrates that the aqueous and ethanolic extracts of D. malabarica seed and flesh could be an effective source of natural antimicrobial agents with no cytotox-ic activity.

Lrp1 in osteoblasts controls osteoclast activity and protects against osteoporosis by limiting PDGF–RANKL signaling

Skeletal health relies on architectural integrity and sufficient bone mass, which are maintained through a tightly regulated equilibrium of bone resorption by osteoclasts and bone formation by osteoblasts. Genetic studies have linked the gene coding for low-density lipoprotein receptor-related protein1(Lrp1) to bone traits but whether these associations are based on a causal molecular relationship is unknown. Here, we show that Lrp1 in osteoblasts is a novel regulator of osteoclast activity and bone mass.Mice lacking Lrp1 specifically in the osteoblast lineage displayed normal osteoblast function but severe osteoporosis due to highly increased osteoclast numbers and bone resorption. Osteoblast Lrp1 limited receptor activator of NF-κB ligand(RANKL) expression in vivo and in vitro through attenuation of platelet-derived growth factor(PDGF-BB) signaling. In co-culture, Lrp1-deficient osteoblasts stimulated osteoclastogenesis in a PDGFRβ-dependent manner and in vivo treatment with the PDGFR tyrosine kinase inhibitor imatinib mesylate limited RANKL production and led to complete remission of the osteoporotic phenotype. These results identify osteoblast Lrp1 as a key regulator of osteoblast-to-osteoclast communication and bone mass through a PDGF–RANKL signaling axis in osteoblasts and open perspectives to further explore the potential of PDGF signaling inhibitors in counteracting bone loss as well as to evaluate the importance of functional LRP1 gene variants in the control of bone mass in humans.

Therapeutic Activity of Partially Purified Fractions of Emblica officinalis （Syn, Phyllanthus embfica） Dried Fruits against Trypanosoma evansi

Emblica officinalis （E. oJficinalis） dried fruits were evaluated for its antitrypanosomal activity and cytotoxic effects. Vero cell line maintained in DMEM （Dubecco＇s Modified Eagle Medium） and incubated with Trypanosoma evansi for more than 12 h. MPE was added to the Vero cell culture medium at different concentrations （250-1,000 μg/mL） with trypanosomes concentration （1 × 106 trypanosomes/mL in each ELISA plate well） and incubated at appropriate conditions for 72 h. In-vitro cytotoxieity of MPE of E. officinalis was determined on Vero cells at concentrations （（1.56-100 ~tg/mL）. Acute toxicity and in-vivo infectivity tests were done in mice. Obtained MPE ofE. officinalis underwent process of purification via column chromatography, preparative chromatography and HPLC （higher performance liquid chromatography） with bioassay at different strata on Alsever＇s medium. In-vivo assay for trypanocidal activity, MPE and PPFs （partially purified fractions） of E. officinalis with two sets of mice, each mouse was inoculated with 1 × 104/mL oftrypanosomes and treated （48 h post inoculation） at concentrations （12.5, 25, 50, 100 and 200 mg/kg body weight） were administered at dose rate of 100 [tL per mouse via intraperitoneal route （in treating parassitemic mice） to different groups of mice, 6 mice per concentration. HPLC of partially purified fractions ofE. officinalis was carried out with mobile phase ofacetonitdle： water （40：60） in gradient mode. In vitro, MPE induced immobilization and killing of the parasites in concentration-time dependent manner. Significant reduction of trypanosomes counts from concentration of 250μg/mL and complete killing of trypanosomes at 5th hour of observation, which was statistically equivalent to 4th hour of Diminazine Aceturate （Berenil）, standard reference drug used. HPLC of the partially purified fractions revealed two major prominent peaks at retention time of 1-4 min. In vivo, both MPE and PPFs of test material did prolong lives of mice by 6-9 days but could not cure them. At concentration of 2,000 kg/kg body weight of MPE in acute test, all mice survived. For in-vivo infectivity test, mice injected with immobilized trypanosomes developed parasitemia and died while, the other group survived. MPE, PPFs and Diminazine Aceturate were toxic to Vero cells at all concentrations exception of 1.56, 1.56-3.13 and 1.56-6.25 μg/mL, respectively. From this report, PPFs ofE. officinalis dried fruits demonstrated potential pathway for a new development oftrypanocide in near future if additional investigations are put in place.

Inhibitory roles of protein kinase B and peroxisome proliferator-activated receptor gamma coactivator on hepatic HMG-CoA reductase promoter activity

Since we had previously demonstrated that siRNAs to tristetraprolin (TTP) markedly inhibited insulin stimulation of hepatic HMG-CoA reductase (HMGR) transcription, we investigated the effects of transfecting rat liver with TTP constructs. We found that transfecting diabetic rats with TTP did not increase HMGR transcription but rather led to modest inhibition. We then investigated whether co-transfection with protein kinase B, hepatic form (AKT2), might lead to phosphorylation and result in activation of HMGR transcription. We found that this treatment resulted in near complete inhibition of transcription. Transfection with peroxisome proliferator-activated receptor g coactivator (PGC-1a) also inhibited HMGR transcription. These results show that although TTP is needed for activation of HMGR transcription, it cannot by itself activate this process. AKT2 and PGC-1a, which mediate the activation of gluconeogenic genes by insulin, exert the opposite effect on HMGR.

不同活动情境下母亲词汇输入特征及其对儿童词汇发展的影响

语言学习的活动情境理论指出,儿童的语言学习嵌入在日常活动情境中。本研究基于这一理论,对汉语儿童与母亲在四种结构化情境下的互动情况进行了分析,发现活动情境、母亲语言特征以及儿童个体特征共同构成了影响汉语儿童词汇语义输入的环境系统。四种结构化活动情境共同推动母亲与儿童互动时产出更加丰富的词汇语义,但不同活动情境下母亲的词汇语义输入具有显著的差异;儿童的年龄和性别特征部分约束了母亲的语言输入特征;活动情境、儿童个体和母亲三个要素在推动儿童语义发展方面都有着不同的作用。研究同时发现,在互动过程中,母亲注重词汇输入的一致性、活动情境性和发展适宜性,但也存在回应敏感度低的问题。基于上述发现,本研究建议关注儿童语言发展的支持性环境系统,特别要营造高质量的结构化活动情境,并进一步提升母亲语言互动质量。

青蒿琥酯纳米胶束制备及其体内药动学、抗肿瘤活性研究

目的制备青蒿琥酯纳米胶束,并考察体内药动学和抗肿瘤活性。方法以聚乙二醇单甲醚-聚乳酸-聚组氨酸(mPEG-PLA-PHis)为载体制备纳米胶束,单因素试验结合Box-Behnken响应面法优化处方,测定包封率、载药量、沉降率、粒径、Zeta电位、体外释药。12只H_(22)荷瘤小鼠随机分为2组,分别尾静脉注射给予青蒿琥酯注射液和青蒿琥酯纳米胶束(1 mg/kg),于不同时间点采血,HPLC法测定青蒿琥酯血药浓度,计算主要药动学参数。36只H_(22)肝癌荷瘤小鼠随机分为6组,即模型组(生理盐水)、阳性组(20 mg/kg环磷酰胺)、青蒿琥酯注射液组(30 mg/kg)及青蒿琥酯纳米胶束低、中、高剂量组(10、20、30 mg/kg),末次给药3 d后记录体质量和瘤重,计算抑瘤率。结果最佳处方为mPEG-PLA-PHis与青蒿琥酯比例10.18∶1,青蒿琥酯质量浓度0.48 mg/mL,水化时间0.96 h,包封率、载药量、沉降率、粒径、Zeta电位分别为(94.27±1.26)%、(8.26±0.18)%、(4.19±0.20)%、(65.14±4.96)nm、-(17.64±1.06)mV。纳米胶束在弱酸性介质中的累积释放度高于在弱碱性介质中,具有pH敏感性。与注射液比较,纳米胶束t_(1/2)、MRT延长(P<0.01),CL降低(P<0.01),AUC_(0~t)升高(P<0.01);与模型组比较,青蒿琥酯纳米胶束不同剂量组小鼠体质量无明显变化(P>0.05),瘤重下降(P<0.05,P<0.01),以中剂量组更明显,抑瘤率达55.40%。结论青蒿琥酯纳米胶束包封率较高,体内抗肿瘤活性较强。

基于光电传感的小动物主动运动行为评价及运动动机的神经机制研究

目的:构建小鼠主动运动行为量化评价系统,为运动与主动健康机制研究、慢病运动干预搭建平台,并通过在体实时采集主动运动不同阶段的脑电信号,探讨运动动机的神经调控机制。方法:1)构建基于光电传感的主动运动多模态系统(voluntary exercise multimodal system, VEMS),包括基于霍尔传感电信号的运动行为采集系统和基于机器视觉的运动轨迹分析系统;2)以C57BL/6小鼠为研究对象,应用VEMS系统记录小鼠主动运动行为并进行量化分析;3)结合在体多通道电生理记录技术采集小鼠主动运动不同阶段中脑腹侧被盖区(ventral tegmental area,VTA)神经元电活动,分析运动动机的神经编码特征。结果:1)VEMS系统可实现小鼠运动节律、运动距离、运动圈数、运动速度、运动时长和运动频次等行为数据的采集及量化分析;2)小鼠日运动时间主要集中在19:00—次日7:00,表现出较强的夜行节律,日运动距离最多可达15 km。每小时最大运动量出现在夜间1:00—2:00,运动圈数为0圈/10 min~170圈/10 min,每小时最大运动频次为23次,速度区间分布在0~12 m/min;3)在运动准备期、运动期及运动停止期,VTA神经元表现出不同的局部场单位(local field potentials,LFPs)特征信号,与运动停止期相比,运动准备期和运动期LFPs信号的θ频段(4~8 Hz)功率谱密度显著升高(P<0.001),且振荡显著增强(P<0.001);4)研究记录到59个对运动特异性响应的多巴胺(dopamine,DA)神经元,平均放电频率为(5.01±0.89) Hz,其中21%的DA神经元(Type-Ⅰ)在运动准备期高频放电,42%的DA神经元(Type-Ⅱ)在运动期有较强放电活动,37%(Type-Ⅲ)在运动停止后放电活动增多(P<0.001)。结论:1)本研究首次创建了VEMS系统,该系统能够对小鼠的主动运动行为进行多维度采集与量化评价分析,不仅为后期精准运动处方制定提供参考模型,还为运动神经科学的研究提供在体实时脑信号采集平台;2)小鼠主动运动行为受到VTA神经元特异性调节,DA神经元在运动准备期、运动期、运动停止期发挥不同作用。Type-Ⅰ和Type-Ⅲ神经元分别在运动准备期和运动停止期双向正反编码,即Type-Ⅰ神经元对奖励性正面信号表现出放电增强,而Type-Ⅲ神经元在运动停止时表现出特异放电。

不同提取工艺桔梗多糖的理化性质分析及其对奥沙利铂体内分布的影响

目的:探究不同提取工艺桔梗多糖(PGP)的理化性质,不同提取工艺所得的PGP对奥沙利铂(OXA)体内分布的影响。方法:本研究采用了热水提取(HW)、醋酸辅助提取(CA)、氨水辅助提取(KA)、超声辅助提取(UA)制备桔梗多糖,通过红外光谱、紫外光谱、凝胶色谱、高效液相色谱等方法测定多糖的理化性质;建立OXA电感耦合等离子体-质谱(ICP-MS)体内分析方法,考察OXA单用组、OXA联用PGP-HW组、OXA联用PGP-CA组、OXA联用PGP-KA组、OXA联用PGP-UA组SD大鼠体内奥沙利铂分布的变化;使用Western Blot方法来测定肺组织中Oct-2和Pg-P蛋白的表达水平,考察PGP改变OXA体内分布的机制。结果:四种桔梗多糖在理化性质方面呈现出明显的区别。其中,PGP-KA的得率最高;四种桔梗多糖红外图谱均具备典型的多糖特征吸收峰;在分子量组成上,它们主要由7 kDa和3790 kDa两个分子量构成,但各自的分子量分布比例有所不同。此外,这四种桔梗多糖都含有甘露糖(Man)、鼠李糖(Rha)、半乳糖醛酸(GalA)、葡萄糖(Glu)、半乳糖(Gal)以及阿拉伯糖(Ara),但比例存在差异;四种多糖粒径主要集中在300 nm左右,且具有良好的溶液稳定性;与OXA单用组比,OXA联用PGP-CA组、OXA联用PGP-KA组分别增加肺组织OXA含量为36.93%和35.12%,且OXA联用PGP-KA组显著(P<0.05)增加Oct-2蛋白表达水平,而P-g-P蛋白表达水平无显著差异。结论:不同的提取工艺获得的PGP的理化性质不同,其中OXA联用PGP-CA组和OXA联用PGPKA组能显著增加OXA在肺组织分布水平,其机制可能与增加肺组织中Oct-2的表达量有关。

In vivo antioxidant activity of rabbiteye blueberry(Vaccinium ashei cv.'Brightwell')anthocyanin extracts

Blueberries are rich in phenolic compounds including anthocyanins which are closely related to biological health functions.The purpose of this study was to investigate the antioxidant activity of blueberry anthocyanins extracted from'Brightwell'rabbiteye blueberries in mice.After one week of adaptation,C57BL/6J healthy male mice were divided into different groups that were administered with 100,400,or 800 mg/kg blueberry anthocyanin extract(BAE),and sacrificed at different time points(0.1,0.5,1,2,4,8,or 12 h).The plasma,eyeball,intestine,liver,and adipose tissues were collected to compare their antioxidant activity,including total antioxidant capacity(T-AOC),superoxide dismutase(SOD)activity and glutathione-peroxidase(GSH-PX/GPX)content,and the oxidative stress marker malondialdehyde(MDA)level.The results showed that blueberry anthocyanins had positive concentration-dependent antioxidant activity in vivo.The greater the concentration of BAE,the higher the T-AOC value,but the lower the MDA level.The enzyme activity of SOD,the content of GSH-PX,and messenger RNA(mRNA)levels of Cu,Zn-SOD,Mn-SOD,and GPX all confirmed that BAE played an antioxidant role after digestion in mice by improving their antioxidant defense.The in vivo antioxidant activity of BAE indicated that blueberry anthocyanins could be developed into functional foods or nutraceuticals with the aim of preventing or treating oxidative stress-related diseases.

芍药甘草汤中6种活性成分在正常、胃溃疡大鼠体内药动学比较

目的比较芍药甘草汤中芍药苷、芍药内酯苷、甘草苷、异甘草苷、甘草素和甘草酸在正常、胃溃疡大鼠体内药动学。方法6只大鼠随机分为2组,建立75%乙醇诱导胃溃疡模型,取胃组织。12只大鼠随机分为2组,灌胃给药(9.9 g/kg),于不同时间点取血,UPLC-MS/MS法测定血药浓度,计算药动学参数。结果与正常组比较,模型组各活性成分T max延长(P<0.05,P<0.01);芍药苷T 1/2、MRT 0~t延长(P<0.05,P<0.01),C max、AUC升高(P<0.05,P<0.01),Vd/F、CL/F降低(P<0.05,P<0.01);芍药内酯苷C max、AUC升高(P<0.05,P<0.01),CL/F降低(P<0.05);甘草苷MRT延长(P<0.05),AUC升高(P<0.05),CL/F降低(P<0.01);异甘草苷MRT延长(P<0.05,P<0.01),Vd/F降低(P<0.05);甘草素药动学参数(除T max外)无明显变化(P>0.05);甘草酸T 1/2、MRT 0~∞延长(P<0.05,P<0.01),C max、AUC升高(P<0.05,P<0.01),CL/F降低(P<0.01)。结论胃溃疡对芍药甘草汤中活性成分体内吸收、代谢的速度和程度有一定影响。

An overview on recent in vivo biological application of cerium oxide nanoparticles

Cerium oxide nanoparticles(CNPs)possess a great potential as therapeutic agents due to their ability to self-regenerate by reversibly switching between two valences+3 and+4.This article reviews recent articles dealing with in vivo studies of CNPs towards Alzheimer’s disease,obesity,liver inflammation,cancer,sepsis,amyotrophic lateral sclerosis,acute kidney injury,radiation-induced tissue damage,hepatic ischemia reperfusion injury,retinal diseases and constipation.In vivo anti-cancer studies revealed the effectiveness of CNPs to reduce tumor growth and angiogenesis in melanoma,ovarian,breast and retinoblastoma cancer cell-induced mice,with their conjugation with folic acid,doxorubicin,CPM,or CXC receptor-4 antagonist ligand eliciting higher efficiency.After conjugation with triphenylphosphonium or magnetite nanoparticles,CNPs were shown to combat Alzheimer’s disease by reducing amyloid-β,glial fibrillary acidic protein,inflammatory and oxidative stress markers in mice.By improving muscle function and longevity,the citrate/EDTA-stabilized CNPs could ameliorate amyotrophic lateral sclerosis.Also,they could effectively reduce obesity in mice by scavenging ROS and reducing adipogenesis,triglyceride synthesis,GAPDH enzyme activity,leptin and insulin levels.In CCl4-induced rats,stress signaling pathways due to inflammatory cytokines,liver enzymes,oxidative and endoplasmic reticulum messengers could be attenuated by CNPs.Commercial CNPs showed protective effects on rats with hepatic ischemia reperfusion and peritonitis-induced hepatic/cardiac injuries by decreasing oxidative stress and hepatic/cardiac inflammation.The same CNPs could improve kidney function by diminishing renal superoxide,hyperglycemia and tubular damage in peritonitis-induced acute kidney injury in rats.Radiation-induced lung and testicular tissue damage could be alleviated in mice,with the former showing improvement in pulmonary distress and bronchoconstriction and the latter exhibiting restoration in spermatogenesis rate and spermatid/spermatocyte number.Through enhancement of gastrointestinal motility,the CNPs could alleviate constipation in both young and old rats.They could also protect rat from light-induced retinal damage by slowing down neurodegenerative process and microglial activation.

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on the Cholesterol-Lowering in Vitro and in Vivo

The survey＇s results showed a significant percentage of women and especially men having an above normal cholesterol. In order to help reduce excessive rate of blood cholesterol, we used a local strain of Spirulina （Tamanrasset, in south of Algeria） associated with probiotic bacteria （Bf adolescentis）. Experiments in vitro showed a significant degradation of total cholesterol by the combination of Spirulina and Bf adolescentis （74.5%） after 72 hours incubation at 37 ~C. A cholesterol is added to the standard diet mice in order to increase the total cholesterol for three lots. However, the rates of total cholesterol in mice receiving lower fermented milk with Bf adolescentis enriched by dry Spirulina. This shows that the decrease of cholesterol rate is closely related to the presence of Bf adolescentis and Spirulina. In vitro and in vivo results show, however, that it is possible to obtain a natural product （Spirulina） and a pseudo-strain lactic （Bf adolescentis） and to participate in the prevention of cardiovascular disease risk factor whose hand is cholesterol.

Downregulation of cold-inducible RNA-binding protein activates mitogen-activated protein kinases and impairs spermatoRenic function in mouse testes

Cold-inducible RNA-binding protein （CIRP） is an RNA-binding protein that is expressed in normal testes and downregulated after heat stress caused by cryptorchidism, varicocele or environmental temperatures. The purpose of this study was to investigate the functions of CIRP in the testes. We employed RNAi technique to knock down the expression of CIRP in the testes, and performed haematoxylin and eosin staining to evaluate morphological changes following knockdown. Germ cell apoptosis was examined by terminal deoxynucleotidal transferase-mediated dUTP nick end labelling （TUNEL） assay, and mitogen-activated protein kinase （MAPK） signalling pathways were investigated by Western blotting to determine the possible mechanism of apoptosis. We found that using siRNA is a feasible and reliable method for knocking down gene expression in the testes. Compared to controls, the mean seminiferous tubule diameter （MSTD） and the thickness of the germ cell layers decreased following siRNA treatment, whereas the percentage of apoptotic seminiferous tubules increased. The p44/p42, p38 and SAPK/JNK MAPK pathways were activated after downregulation of CIRP. In conclusion, we discovered that downregulation of CIRP resulted in increased germ cell apoptosis, possibly viathe activation of the p44/p42, p38 and SAPK/JNK MAPK pathways.

黄秋葵抗氧化活性研究

目的研究黄秋葵果实不同极性部位提取物的体外和体内抗氧化能力,为黄秋葵的产品开发和利用提供依据。方法采用·DPPH清除能力、·ABTS^(+)清除能力、·OH清除能力、·O_(2)^(-)清除能力、FRAP法还原能力、TBARS法抗脂质过氧化能力体外抗氧化模型对黄秋葵果实95%乙醇提取物中不同极性部位进行抗氧化活性评价,并利用SPSS 19.0软件分析黄秋葵果实抗氧化活性与总黄酮含量之间的相关性。选择其具有体外抗氧化活性部位的提取物进行体内抗氧化活性研究。结果95%乙醇提取物中乙酸乙酯层和正丁醇层提取物表现出清除自由基能力、还原能力和抑制脂质过氧化能力,乙酸乙酯层和正丁醇层提取物的总黄酮含量高于其他各层提取物,总黄酮含量与抗氧化活性呈较强相关性。与模型组比较,给予乙酸乙酯层和正丁醇层提取物能够升高CCl_(4)肝损伤大鼠肝脏中SOD、CAT活性以及GSH水平,同时能够降低ALT活性和MDA水平。结论黄秋葵果实具有潜在的护肝作用,可为开展肝脏损伤保护研究提供重要依据。

Antioxidant Activities of Crude Phlorotannins from Sargassum hemiphyllum

Brown algae are well known as a source of biologically active compounds, especially those having antioxidant activities, such as phlorotannins. In this study we examined the antioxidant activities of crude phlorotannins extracts（CPEs） obtained from Sargassum hemiphyllum（SH） and fractionated according to the molecular weights. When CPEs were administrated at a dose of 30 mg/kg to Kunming mice pre-treated with carbon tetrachloride（CCl4）, the levels of oxidative stress indicators in the liver, kidney and brain were significantly reduced in vivo. All the components of various molecular weight fractions of CPEs exhibited greater scavenging capacities in clearing hydroxyl free radical and superoxide anion than the positive controls gallic acid, vitamin C and vitamin E. Particularly, the components greater than 30 k D obtained from ethyl acetate phase showed the highest antioxidant capacities. These results indicated that SH is a potential source for extracting phlorotannins, the algal antioxidant compounds.