BioNoculars Extracting Protein-Protein Interactions from Biomedical Text

Protein A/G免疫沉淀磁珠Figure 1. General Protocol for ImmunoprecipitationcomplexSDS-PAGE loading buffer Neutralize bufferMagnetic Beads antibodyMagnetic Separator Remove supernatant Pipette Repeat45产品组分产品参数：磁珠粒径100 nm，浓度10 mg/mL，结合量>400 μg human IgG/mL2-8℃保存，保质期2年。

储存方法实验步骤1. 抗原样品制备本操作说明书提供以下三种样品处理方法。

2. 磁珠预处理将磁珠漩涡振荡1 min，使其充分混悬；取25~50 µL磁珠悬液置于1.5 mL EP管中。

加入200 µL结合缓冲液洗涤，进行磁性分离（将离心管置于磁力架上，管底对准①卡口压紧，静置2分钟或待磁珠吸附于管壁），吸弃上清。

抽出②磁条，加入200 µL结合缓冲液重复洗涤一次，插回②磁条，磁性分离并吸弃上清。

加入200 µL结合缓冲液重悬磁珠备用。

血清样品处理：若目标蛋白丰度较高， 建议用结合缓冲液稀释血清样品至目标蛋白终浓度为10~100 µg/mL，置于冰上备用（或置于-20℃长期保存）。

悬浮细胞样品处理：离心收集细胞（4℃, 500 g, 10 min），弃上清后称重，按每毫克细胞50 µL的比例用1×PBS洗涤2次；按每毫克细胞5~10 µL的比例加入结合缓冲液，同时加入蛋白酶抑制剂，混匀后置于冰上处理10 min；离心收集上清液（4℃, 14000 g, 10 min），置于冰上备用（或置于-20℃长期保存）。

贴壁细胞样品处理：移去培养基，按每1.0×105个细胞150 µL的比例用1×PBS洗涤两次；用细胞刮棒刮脱细胞，收集至1.5 mL EP管内，按每1.0×105个细胞20~30 µL的比例加入结合缓冲液，同时加入蛋白酶抑制剂，混匀后置于冰上处理10 min；离心收集上清液（4℃, 14000 g, 10 min），置于冰上备用（或置于-20℃长期保存）。

桑黄提取物体内抗肿瘤作用的实验研究
实验采用小鼠肺癌细胞C126的体内植入瘤模型。

将小鼠随机分为对照组和实验组，实验组分为三个剂量组，每组6只小鼠。

实验组给予桑黄提取物不同剂量的腹腔注射，每周
注射三次，连续注射4周，对照组给予等量的生理盐水腹腔注射。

观察小鼠体重变化、肝
和肾功能及脾脏、心脏和肺组织病理学变化情况。

同时通过免疫细胞检测、MTT法和流式
细胞仪检测桑黄提取物的抗肿瘤作用。

结果显示，桑黄提取物能够抑制小鼠肺癌细胞的生长，具有显著的抗肿瘤作用。

其中，高剂量组的肿瘤抑制率最高，为61.2%。

在瘤组织中观察到大量的细胞凋亡和坏死现象，
说明桑黄提取物具有诱导肺癌细胞凋亡的作用。

同时，实验组小鼠的体重、肝和肾功能、
脾脏、心脏和肺组织都没有明显的毒副作用和病理学变化。

此外，桑黄提取物还能够明显提高小鼠免疫系统的活性，增强机体的免疫功能。

MTT
法的结果表明，桑黄提取物对小鼠淋巴细胞增殖有促进作用。

流式细胞仪检测显示，桑黄
提取物能够提高小鼠血清中白细胞、淋巴细胞和NK细胞的数量，同时减少调节性T细胞的数量。

综上所述，桑黄提取物具有良好的抗肿瘤作用，能够抑制小鼠肺癌细胞的生长，同时
具有提高小鼠免疫系统活性的作用。

本研究结果为桑黄的临床应用提供了一定的科学依据
和实验基础。

《国际商务标准水飞蓟提取物》编制说明1 任务来源本标准的制定工作，是由中国医药保健品进出口商会提出而进行的，国际商务标准植物提取物编号为WM 。

本标准由长沙康隆生物制品有限公司与盘锦天源药业有限公司共同起草。

2 标准制定的意义水飞蓟提取物是由菊科植物水飞蓟(Silybum marianuml(L.)Gaertn.) 的干燥成熟果实中提取得到的黄酮类化合物,主要包括水飞蓟亭、水飞蓟宁、水飞蓟宾、异水飞蓟宾等。

水飞蓟宾(Silybin) 为主要有效成分, 包括水飞蓟宾A和水飞蓟宾B。

溶于丙酮、乙酸乙酯、甲醇及乙醇，不溶于水。

它具有保护肝脏、改善肝功能、增强肝细胞再生等作用，对急慢性肝炎、肝硬化及代谢中毒性肝损伤等均有较好疗效。

水飞蓟提取物含量的测定主要按水飞蓟宾计。

水飞蓟提取物是用于治疗肝脏疾病的常用药物。

药理作用表明水飞蓟提取物主要是通过限制ROS（reactive oxygen species）的活性来实现其治疗作用的，也常被用于保健食品中。

美国药典USP35–NF30 中水飞蓟提取物（Powdered Milk Thistle Extract）和欧洲药典（European pharmacopoeia 7.0）中水飞蓟提取物（MILK THISTLE DRY EXTRACT,REFINED AND STANDARDISED）中都有关于水飞蓟提取物的技术指标及检验标准，《中华人民共和国药典》2010版一部上收录了水飞蓟标准，但是国内并没有关于水飞蓟提取物的完善标准依据。

当前外贸出口贸易中的食品安全形式十分严峻，为保障国家外贸经济运行的安全和我国人民群众的食品卫生安全，加强标准建设，促进与国际水飞蓟提取物标准接轨，及时建立水飞蓟提取物国际商务标准的国内质控标准具有重要的现实意义。

3 标准编写规则本标准遵循GB/T1.1-2009《标准化工作导则第1部分：标准的结构和编写规则》；GB/T20001.2-2001《标准化工作指南第2部分：采用国际标准的规则》和GB/T20001.4-2001《标准编写规则第4部分：化学分析方法》规则编写。

植物组织蛋白质提取方法1、植物组织蛋白质提取方法1、根据样品重量(1g样品加入3.5ml提取液,可根据材料不同适当加入),准备提取液放在冰上。

2、把样品放在研钵中用液氮研磨,研磨后加入提取液中在冰上静置(3-4小时)。

3、用离心机离心8000rpm40min4℃或11100rpm20min4℃4、提取上清夜,样品制备完成。

蛋白质提取液:300ml1、1Mtris-HCl(PH8)45ml2、甘油(Glycerol)75ml3、聚乙烯吡咯烷酮(Polyvinylpolypyrrordone)6g这种方法针对SDS-PAGE,垂直板电泳!2、植物组织蛋白质提取方法三氯醋酸—丙酮沉淀法1、在液氮中研磨叶片2、加入样品体积3倍的提取液在-20℃的条件下过夜,然后离心(4℃8000rpm 以上1小时)弃上清。

3、加入等体积的冰浴丙酮(含0.07%的β-巯基乙醇),摇匀后离心(4℃8000rpm以上1小时),然后真空干燥沉淀,备用。

4、上样前加入裂解液,室温放置30分钟,使蛋白充分溶于裂解液中,然后离心(15℃8000rpm 以上1小时或更长时间以没有沉淀为标准),可临时保存在4℃待用。

5、用Brandford法定量蛋白,然后可分装放入-80℃备用。

药品:提取液:含10%TCA和0.07%的β-巯基乙醇的丙酮裂解液:2.7g尿素0.2gCHAPS溶于3ml灭菌的去离子水中(终体积为5ml),使用前再加入1M的DTT65ul/ml。

这种方法针对双向电泳,杂质少,离子浓度小的特点!当然单向电泳也同样适用,只是电泳的条带会减少!3、组织:肠黏膜目的:WESTERN BLOT检测凋亡相关蛋白的表达应用TRIPURE提取蛋白质步骤:含蛋白质上清液中加入异丙醇:(1.5ml每1mlTRIPURE用量)倒转混匀,置室温10min离心:12000 g,10min,4度,弃上清加入0.3M盐酸胍/95%乙醇:(2ml每1mlTRIPURE用量)振荡,置室温20min离心:7500g,5 min,4度,弃上清重复0.3M盐酸胍/95%乙醇步2次沉淀中加入100%乙醇2ml充分振荡混匀,置室温20 min离心:7500g,5min,4度,弃上清吹干沉淀1%SDS溶解沉淀离心:10000g,10min,4度取上清-20度保存(或可直接用于WESTERN BLOT)存在的问题:加入1%SDS后沉淀不溶解,还是很大的一块,4度离心后又多了白色沉定, SDS结晶?测浓度,含量才1mg/ml左右。

网络出版时间：２０２４－０１－１０１０：５８：４０　网络出版地址：ｈｔｔｐｓ：／／ｌｉｎｋ．ｃｎｋｉ．ｎｅｔ／ｕｒｌｉｄ／３４．１０８６．Ｒ．２０２４０１０８．１８３１．０３８◇网络药理学◇液相色谱－质谱联用技术分析秦巴硒菇提取物活性成分及其治疗慢性粒细胞白血病的网络药理学研究王东萍１，４，葛万文２，邵　晶３，孙延庆１，４（１．甘肃中医药大学中西医结合学院，甘肃兰州　７３００００；２．兰州大学第二医院，甘肃兰州　７３００３０；３．甘肃中医药大学药学院，甘肃兰州　７３００００；４．甘肃省人民医院，甘肃兰州　７３００００）收稿日期：２０２３－０９－２０，修回日期：２０２３－１１－２１基金项目：国家自然科学基金资助项目（Ｎｏ８１５６０６７０）；甘肃省自然科学基金资助项目（Ｎｏ２０ＪＲ１０ＲＡ３７６，２１ＪＲ１１ＲＡ１９６）；甘肃省人民医院国家级科研项目培育计划（１９ＳＹＰＹＢ １７）；兰州市科技发展指导性计划项目（Ｎｏ２０２０ ＺＤ ５６）作者简介：王东萍（１９８６－），女，硕士，研究方向：中西医结合血液病，肿瘤药理学，Ｅ ｍａｉｌ：ｗａｎｇｄｐ０８３１＠ｇｓｚｙ．ｅｄｕ．ｃｎ；孙延庆（１９６４－），男，博士，教授，主任医师，博士生导师，研究方向：中西医结合血液病，肿瘤药理学，通信作者，Ｅ ｍａｉｌ：４０ｙａｎｑｉｎｇｆａｎｇ＠ｇｓｚｙ．ｅｄｕ．ｃｎｄｏｉ：１０．１２３６０／ＣＰＢ２０２３０３０２８文献标志码：Ａ文章编号：１００１－１９７８（２０２４）０１－０１３９－０７中国图书分类号：Ｒ２５８ ５；Ｒ３１９；Ｒ４４６ ９；Ｒ７３３ ７摘要：目的　利用液相色谱质谱联用和网络药理学、分子对接技术探讨秦巴硒菇提取物治疗慢性粒细胞白血病（ｃｈｒｏｎｉｃｍｙｅｌｏｉｄｌｅｕｋｅｍｉａ，ＣＭＬ）的潜在活性靶点及相关信号通路，并通过体外实验进一步验证。

方法　应用液相色谱质谱分析秦巴硒菇提取物的活性成分，通过ＳｗｉｓｓＴａｒｇｅｔＰｒｅｄｉｃｔｉｏｎ数据库预测药物靶点；从ＧｅｎｅＣａｒｄｓ、ＤｉｓＧｅＮＥＴ数据库获取ＣＭＬ的疾病靶点。

贵州省遵义市2023-2024学年高一下学期6月月考英语试卷一、阅读理解Covering over 1,600 square kilometers of England’s most valued lowland landscapes (风景) in the busiest part of the UK, the South Downs National Park has been shaped by the activities of its farmers and foresters, its charities and local businesses. Find out about some events happening across the park.Benfield Hill City Nature ChallengeIf you would like to be part of the global City Nature Challenge which brings together cities and organizations around the world to share observations of nature, we will be holding our own initiative on Saturday, 27th May, on Benfield Hill Local Nature Reserve. Welcome anyone, whatever their level of experience, in supporting us on a fun day of learning and identification of valuable biodiversity.Green Sketching WorkshopDiscover how you can use the process of drawing to look at and notice nature, and as a tool for slowing down and bringing calm to our busy lives. This is not a how-to-draw workshop, but a how-to-see workshop! This focuses on the process of drawing rather than the finished result, which means that everyone, regardless of previous drawing experience, can benefit from the joy of Green Sketching.Longmoor Through the AgesDiscover more about this vast land and how humans have shaped the landscape around the site. Bring your binoculars (双筒望远镜) as the site is also part of the Wealden Heath Phase Ⅱ Special Protection Area and home to some rare (珍稀的) birds, reptiles and rare species of international importance.Dawn Chorus WalkGet up with the birds. You won’t regret setting your alarm as we enjoy the magic of some of our springtime songsters. Shortheath Common sits at the northern extremity of the South Downs National Park and is regarded as a Special Area of Conservation due to the unique ecological landscape. It’s hope to a variety of rare birds, and plant species of international importance. 1．Which of the following most attracts people who want to use painting to show nature?A．Benfield Hill City Nature Challenge.B．Green Sketching Workshop.C．Longmoor Through the Ages.D．Dawn Chorus Walk.2．What can people do at Longmoor Through the Ages?A．Hold meetings.B．See painting exhibitions.C．Record the farmers’ songs.D．Watch some rare animals.3．What is the purpose of the text?A．To tell about the history of the South Downs National Park.B．To encourage donations to the South Downs National Park.C．To stress the importance of the South Downs National Park.D．To introduce activities happening across the South Downs National Park.In 2019, after retiring from her career as a social worker, Ane Freed -Kernis decided to build a home workshop and devote all of her free time to stone carving. “I might be covered head to to e in dust but I’m happy — it was something I needed more of in my life when I hit 60,” she says.This appeal has its origins in Freed - Kernis’ childhood. Growing up on her father’s farm in Denmark, she used to wander through the fields with her eyes fixed on the ground, looking for stones to add to her collection. “I’ve always been drawn to the shapes and textures(质地) of stones,” she says.After moving to England in 1977 and training as a social worker, Freed -Kernis soon became occupied with her busy career and the demands of raising her son. Stones were the last thing on her mind, until her father died in 2005. “He took a stone carving course in his retirement, and I always thought stone seemed so fun but never had the time to look into it myself,” she says. “After he died, I became determined to learn in his honour.”Signing up for a week-long stone carving course at Yorkshire Sculpture Park, Freed -Kernis began to learn how to turn a block of rock into well-designed shapes. “It was really scary at the start because you would spend hours just hammering(锤打).”Now 65, Freed - Kernis has a thriving small business built largely through word of mouth. She creates 12 to 15 pieces a year that can take anywhere from a few days to three weeks to complete, while her prices range from £ 200 to £ 3,000. “I’m making smaller ones,” she says. “I don’t have to depend on the money much, so I want to keep prices in the range that people can afford, mainly just covering costs and labour(劳动力).”4．Freed-Kernis was first attracted by stones when ______.A．she was 60B．she was a childC．her father died D．she moved to England5．What can we infer about Freed-Kernis from paragraph 3?A．She never cared about her father.B．She led a disappointing life in Denmark.C．She spent lots of time studying stone carving.D．She learned stone carving under the influence of her dad.6．How did Freed-Kernis feel when she started stone carving course?A．Hopeful and proud.B．Confident and satisfied.C．Nervous and frightened.D．Impatient and unprepared.7．Why is Freed-Kernis making smaller pieces?A．They are easier to move by her.B．They are more affordable to people.C．She wants to save costs and labour.D．She is too old to focus on making large ones.In San Francisco, a large group of sea lions move themselves out of the bay waters and hang out on PIER 39, which is a popular tourist destination. According to dock (码头)officials, this is the most sea lions seen in the region in 15 years.“Over 1,000 sea lions have been counted this week,” PIER 39 harbormaster Sheila Chandor told many different media. “The surge in sea lions is usually a good sign of their strong population and healthy living environment,” said Adam Ratner, Director of Conservation Engagement at the Marine Mammal(海洋哺乳动物) Center in Sausalito, California.“California sea lions are sentinels(哨兵) of the ocean,” Ratner said. Their population to some extent reflects the health of the ocean. Therefore, seeing a large number of California sea lions is clearly a good thing.For nearly 35 years, the slippery(滑的) residents have been a star attraction for tourists. That autumn in 1989, PIER 39 had just been repaired, but the ships had not yet been moved back.At that moment, the sea lions unexpected arrival not only attracted fans but also created enemies. According to a website, some dock residents and workers were scared away by the strong and very unpleasant smell and noise of their new neighbors, while others saw these animals as a bright spot after the destructive Loma Prieta earthquake.The officials sought help from the Marine Mammal Center to find a way to deal with sea lions. Ratner said that the final decision is to let the sea lions stay and coexist with humans. “The fact proves that this is really a good thing,” he said. “This is just a proof of how we can truly work together and think about how we can share our coasts with marine mammals and other wildlife in a way that benefits all the parties involved.”8．How does the author start the text?A．By describing a situation.B．By answering a question.C．By holding a conversation.D．By comparing different opinions.9．What does the underlined word “surge” in paragraph 2 mean?A．Sharp increase.B．Tight control.C．Slow development.D．Sudden movement.10．What is Ratner’s attitude to the final decision?A．Doubtful.B．Uninterested.C．Supportive.D．Unclear.11．What message does the author seem to convey in the text?A．Sea lions are pretty cool animals.B．Animals and humans can live in harmony.C．Watching sea lions might not be a proper action.D．Sea lions should be driven out of PIER 39.With mounting evidence that nanoplastic particles (纳米塑料微粒) are in our bodies, there is growing concern over their potential health impacts. Now a new study finds a relation between nanoplastics in the brain and a higher risk for Parkinson’s disease.Nanoplastics appear when the plastic packaging breaks down into small pieces. Theseparticles can enter the blood and cross the blood-brain barrier, with European researchers reporting earlier this year that in animal experiments, it can take two hours or less for certain nanoplastics to reach the brain after being eaten.In humans, it’s long been thought that environmental factors play a role in Parkinson’s disease but specific causes are still unclear. The new study from the Duke University School of Medicine details how nanoplastics cause chemical changes in the brain that can, in turn, make Parkinson’s and related types of diseases more likely.That’s because the nanoplastics attract a protein (蛋白质) called alpha-synuclein, known to play a role in Parkinson’s and related disorders. In lab and animal studies, the plastic’s interaction with it leads to increases in the affected neurons in the brain. This interaction appears related to favorable conditions in which Parkinson’s can develop.The study authors note that Parkinson’s disease existed long before nanoplastics appeared in the environment, but they think that this “nanoplastics pollution in the human brain” may prove a new poison.Further, the Duke team led by Dr. Andrew West notes that Parkinson’s disease is among the fastest growing nervous diseases in the world, even as the amazing amount of plastic pollution builds across the planet. This is expected to continue for the foreseeable future.“The technology to monitor nanoplastics is still at the earliest possible stages and not ready yet to answer all the questions we have,” West said. “But hopefully efforts in this area will increase rapidly, as we see what these particles can do in our experiments.”12．Where is the text most probably taken from?A．A product advertisement.B．A science journal.C．An art magazine.D．A travel brochure.13．What is paragraph 4 mainly about?A．The conditions leading to Parkinson’s.B．The cause of alpha-synuclein’s appearance.C．The principle of nanoplastics’ impact on Parkinson’s.D．The difference between Parkinson’s and related disorders.14．What can be inferred from West’s words in the last paragraph?A．Plastic pollution will by no means be avoided.B．Nanoplastics are impossible to deal with at present.C．Fewer people will suffer from Parkinson’s in the future.D．More efforts in the study of nanoplastics will be put in.15．What is a suitable title for the text?A．Nanoplastics can enter the brain through bloodB．Nanoplastics may promote Parkinson’s diseaseC．Alpha-synuclein plays a role in Parkinson’s diseaseD．Nanoplastics will do serious harm to human healthRoommates can provide support, creating a shared space where memories are made and challenges are faced together. 16 , but it takes some efforts to make it work. Here are some tips for living peacefully with roommates.Establish boundaries (界限)17 . But experts suggest being careful when moving in with them, as it could do harm to the relationship if you’re not clear about boundaries. So being clear about boundaries is one way of making sure everyone feels comfortable.Ask hard questionsHave a discussion of your living arrangement. How are you paying rent? What’s your guest policy (政策)? 18 ? And make your expectations clear. If your roommates say they can pay rent on time, for example, you can tell them the specific time to pay rent. 19In a shared living arrangement, it’s perfect for everyone to feel free to express their concerns and opinions without fear of judgment (判断). To do this, encourage open-mindedness and active listening. Avoid making assumptions (假设), forcing your opinions on others and not thinking about others’ views.Learn from themLiving with a roommate is a unique opportunity to meet someone with a very different background from yours. 20 , showing appreciation and giving within your ability do so much for the relationship and create a space with love.A．Where do you come fromB．Figure out your shared interestC．Should you create a cleaning scheduleD．Even if you’re not best friends with themE．Create a free and non-judgmental environmentF．Having a good roommate can be a great experienceG．It’s okay to share space with friends who are polite and responsible二、完形填空Juleus Ghunta and his three sisters lived in a rural part of Western Jamaica. They were 21 by a single mother, and his mother often had to make 22 choices about how to use their limited resources-including a decision to send his oldest sister to school, but to 23 Ghunta at home.When Ghunta finally went to school, he couldn’t catch up on his reading skills. “I 24 in school with a deep sense of loss and 25 ,” he said. Not only had he been kept home from school as a child, but he had not been exposed (使接触) to 26 .When Ghunta was about 12, a young teacher decided to start a special 27 program for struggling students. Ghunta was the first student to 28 . “The teacher was 29 kind to me.” he said. “She was patient. She did not require anything of me, except that I 30 in myself and work hard.” Under her 31 , Ghunta’s reading skills finally started to improve. And his sense of inadequacy (能力不足) 32 to lift.After Ghunta’s experience with the teacher, his life 33 a new direction. He went on to college, and later, graduate school. Today, he is the author of two children’s books.He would like to thank his teacher for seeing his 34 . “I would love for her to see the significant 35 that she has made on my life, and how it continues to be a source of joy.”21．A．monitored B．replaced C．observed D．raised 22．A．tough B．annoying C．confusing D．familiar 23．A．contact B．comfort C．keep D．compare 24．A．lived B．adapted C．volunteered D．struggled 25．A．responsibility B．shame C．humour D．achievement 26．A．families B．books C．neighbours D．friends27．A．reading B．experiment C．writing D．fitness 28．A．build up B．mix up C．sign up D．make up 29．A．partly B．hardly C．suddenly D．extremely 30．A．check B．believe C．take D．result 31．A．control B．protection C．guidance D．consideration 32．A．began B．decided C．failed D．chose 33．A．closed B．doubted C．feared D．took 34．A．memories B．possibilities C．explanations D．instructions 35．A．reasons B．summaries C．impacts D．challenges三、语法填空阅读下面短文，在空白处填入1个适当的单词或括号内单词的正确形式。

PROTEIN EXTRACTION KIT DIAGENODEContentIntroduction 4 General remarks before starting 4 Required materials and reagents 5 Precautions 5 Protocol 6 |PROTEIN EXTRACTION KITPAGE 4 DIAGENODE M A N U A L Innovating Epigenetic SolutionsIntroductionProtein extraction from tissues and cultured cells is the first step for many biochemical and analytical techniques (PAGE, Western blotting, mass spectrometry, etc.) or protein purification. Efficient disruption and homogenization of animal tissues and cultured cells are required to ensure high yields of proteins. Diagenode’s Bioruptor ® uses state-of-the-art ultrasound technology to efficiently disrupt and homogenize tissues and cultured cells in just one step. Bioruptor ® offers unique benefits for tissue disruption and homogenization:• Fast and simple• No contamination between samples• Efficient• Gentle processing• Reproducible• Temperature controlled • Multiplexing capability General remarks before starting• Conditions for protein extraction (e.g. use of fresh or frozen tissue, composition of extraction buffer etc.) must be adjusted according to the nature of the proteins of interest and the assays to be run. SDS might be added to the extraction buffer to maximize the yield of soluble proteins. SDS extracts can be used for SDS electrophoresis and Western blotting. It is recommended to reduce the SDS concentration for 2D electrophoresis, enzyme-linked immunosorbent assay and mass spectrometry.• For functional studies (e.g. the study of protein–protein interactions), avoid using ionic detergents and high concentrations of salt.Optional: use RIPA buffer as a starting point for optimization:50 mM Tris-HCl (pH 7.4) 150 mM NaCl 1% NP-40 0.25% Na-deoxycholateProtease Inhibitor Mix SDS 0.1 - 2% (optional)It is always recommended to optimize the buffer composition depending on a specific research project• Always use Protease Inhibitor Mix during extraction procedure to block the possible protein degradation.• Use Diagenode’s TPX tubes for sonication. Depending on the desired final volume, 1.5 ml TPX microtubes (Diagenode, Cat. No.: M-50050 or M-50001) or 15 ml TPX tubes (Diagenode, Cat. No.: M-UN-15) might be used. Always respect the recommended sonication volumes: 100 - 300 µl for 1.5 ml TPX tubes and 1 - 2 ml for 15 ml TPX tubes (strictly follow the Bioruptor ® instructions as shown in the corresponding manual before starting any sonication experiments).• Keep extracted proteins at -80°C. PAGE 5 |PROTEIN EXTRACTION KIT DIAGENODE Required materials and reagents• Bioruptor ® Standard or Plus (Diagenode, Cat. No. UCD-200, UCD-300)• Bioruptor ® Water Cooler (Diagenode, Cat. No. BioAcc-Cool)• Single Cycle Valve for Bioruptor ® Plus (Diagenode, Cat. No. VB-100-0001)• 1.5 ml (Diagenode, Cat. No. M-50050 or M-50001) or 15 ml TPX tubes (Diagenode, Cat. No. M-UN-15) for sonication • Protein extraction kit (Diagenode, Cat. No. AL-002-0050)• Protein Extraction Beads (Diagenode, Cat No. AL-003-0020) for tissue disruption (not required for cell lysis)• Tube holder for 1.5 ml tubes (Diagenode, Cat. No. UCD-pack 1.5)• Tube holder pack for extraction kits (Diagenode, Cat. No. O-ring-15)• Protease Inhibitor Mix (Diagenode, Cat. No. kch-502-300 or kch-502-100)• Buffer for protein extraction from tissue or cell lysis (not supplied)• Reagents for protein quantification (optional)Precautions• This product is intended for laboratory research use only. CAUTION : Not for diagnostic use. The safety and efficacy of this product in diagnostic or other clinical uses has not been established. • RNA extraction reagent is harmful and not intended for medical use. Handle with great care. • When using, wear appropriate protective gear (gloves, goggles, etc.). • If the product enters eye or adheres to skin, wash with large amounts of water for at least 15 minutes and consult a doctor. • Handle this product in accordance with the manual. • Diagenode is not responsible for problems caused if this product is not handled in accordance with the manual.PROTEIN EXTRACTION KITPAGE 6 DIAGENODE M A N U A L Innovating Epigenetic SolutionsProtocolI. Protein extraction from Tissues »This protocol has been validated for up to 50 mg of tissue. Do not use more tissue per sample. For larger quantity cut the tissue and proceed to the disruption in separate tubes. When proceeding 20 - 50 mg of tissue 15 ml TPX tubes are recommended with a final volume of 1 - 2 ml. Less tissue could be sonicated in 1.5 ml TPX tubes with a final volume of 100 - 300 µl. »Minimize the time of tissue collection to prevent protein degradation.»Dissected tissues can be snap-frozen in liquid nitrogen and stored at -80°C until protein extraction1. Pre-cool Bioruptor ® to 4°C using the Bioruptor ® Water Cooler (Diagenode, Cat. No. BioAcc-Cool).2. Fill the TPX tubes with Protein Extraction Beads.  »The recommended quantity of the beads is 200 - 250 mg for 15 ml TPX tubes, 40 - 50 mg for 1.5 ml TPX tubes. Note: If using pre-filled tubes (Cat. No. AL-002-0050 Protein Extraction kit) please skip this step!3. Add Protease Inhibitor Mix (200x) to the cold protein extraction buffer: 5 µl per 1 ml of extraction buffer. Scale accordingly. 4. Add the required volume of a cold extraction buffer to the TPX tubes pre-filled with Protein Extraction Beads. 5. Add tissue pieces to the TPX tubes. Make sure that the final volume is in the recommended range: 100 - 300 µl for 1.5 ml TPX microtubes and 1 - 2 ml for 15 ml TPX tubes. 6. Vortex tubes briefly and proceed to sonication by using the Bioruptor ® with the following settings:Power: H position (High) Sonication cycle: 30 sec ON/30 sec OFF Total sonication time: 5 - 15 cycles Temperature: 4°C »To guarantee homogeneity of sonication, the tube holder should be always completely filled with tubes.7. Stop the Bioruptor ® after each 5 cycles, vortex samples and check the sample visually for disruption. »Please note that the optimization might be required depending on the sample format (fresh or frozen tissue), tissue type and tissue amount. The shortest sonication time should be chosen to prevent protein damage. Incomplete disruption may occur with fibrous tissues (i.e. muscles). 8. Transfer the supernatant to a new tube and centrifuge samples at 14,000 rpm for 15 min at 4°C to remove any remaining insoluble material.»The Protein Extraction Beads might be washed once with extraction buffer for maximum recovery of total protein but this will lead to the sample dilution. 9. Transfer the supernatant containing soluble proteins to a new tube. 10. Take an aliquot for the quantification and the further analysis if needed. Store proteins extracts in small aliquots at -80°C.»Different protein concentration assays exist including: absorbance at 280 nm, Lowry Assay, Bradford Assay, Bicinchoninic Assay (BCA) etc.. Many commercial kits for protein quantification are also available. Please note that measuring the protein concentration in an SDS extract requires that the assay is compatible with the detergent and reducing agent in the solution. PAGE 7 |PROTEIN EXTRACTION KIT DIAGENODE II. Protein extraction from Cultured Cells»This protocol has been validated using RIPA buffer but it may be necessary to optimize the buffer composition depending on a specific research project. »We recommend using 100 µl of an appropriate lysis buffer per 1x10^6 cells. »For Western blotting, cells might be lysed directly in 1x Laemmli buffer. After sonication, centrifuge extract at 14,000 rpm for 15 min. Transfer the supernatant to a new tube and boil for 3 min. The supernatant can be used in Western blot. Note that protein quantification by common methods is not compatible with Laemmli buffer.1. Pre-cool Bioruptor ® to 4°C using the Bioruptor ® Water Cooler (Diagenode, Cat. No. BioAcc-Cool).2. A dd Protease Inhibitor Mix (200x) to the ice-cold cell lysis buffer: 5 µl per 1 ml of extraction buffer. Scale accordingly. 3. For monolayer cells: R inse the monolayer cells 3 times with cold PBS. For the final rinse, use a cell scraper and transfer the cell suspension to a TPX tube. Centrifuge cells at 1,500 rpm for 10 min at 4°C and aspirate as much supernatant as possible. Proceed to step 4. For suspension cells: C entrifuge suspension at 1,500 rpm for 10 min at 4°C and aspirate the supernatant. Resuspend the pellet in cold PBS, transfer to a TPX tube and centrifuge at 1,500 rpm for 10 min at 4°C. Aspirate the supernatant. Repeat 2 more times. Proceed to the step 4. 4. Add ice-cold cell lysis buffer and resuspend the pellet. Incubate on ice for 10 min.»The viscosity may appear at this step5. Vortex tubes briefly and proceed to sonication by using the Bioruptor ® with the following settings:Power: H position (High) Sonication cycle: 30 sec ON/30 sec OFF Total sonication time: 5-10 cycles Temperature: 4°C »To guarantee homogeneity of sonication, the tube holder should be always completely filled with tubes.6. Stop the Bioruptor ® after 5 cycles, briefly vortex samples and visually check the samples: Samples should be in solution (viscosity should be reduced)»Please note that the optimization might be required depending on sample format (cell density, cell type etc.). The shortest sonication time should be chosen to prevent protein damage. 7. Transfer the supernatant to a new tube and centrifuge samples at 14,000 rpm for 15 min at 4°C to remove any remaining insoluble material.8. Take an aliquot for the quantification and the further analysis if needed. Store protein extracts at -80°C.»Different protein concentration assays exist including: absorbance at 280 nm, Lowry Assay, Bradford Assay, Bicinchoninic Assay (BCA) etc. Many commercial kits for protein quantification are also available. Please note that measuring the protein concentration in an SDS extract requires that the assay is compatible with the detergent and reducing agent in the solution.PROTEIN EXTRACTION KITPAGE 8 DIAGENODE M A N U A LInnovating Epigenetic SolutionsFigure 1. Protein Extraction Beads are required for efficient tissue disruption using the Bioruptor ®Complete disruption is observed in the sample containing Diagenode’s Protein Extraction Beads (left) after 5 cycles while non-disrupted tissue is still present in the sample without the Protein Extraction Beads (right).Figure 2. Total proteins effectively extracted from tissues using Bioruptor ®Various mouse tissues were disrupted in RIPA buffer supplemented with or without 2% SDS. Total proteins were separated by SDS-PAGE and stained with Coomassie Blue dye.Liver Brain Muscle + SDS + SDS + SDS - SDS- SDS- SDSPAGE 9 |PROTEIN EXTRACTION KIT DIAGENODE Figure 3. Western blot analysis of GAPDH and HSP90 proteins in tissue and cultured cell extracts.Expected bands of 37 kD and 90 kD are observed for GAPDH (left panel) and HSP90 (right panel), respectively, in liver, brain and skeletal muscle. Note that HSP90 is expressed in muscle in an extremely low level (H. Quraishi and I. R. Brown, J Neurosci Res. 1996 Feb 1;43(3):335-45). Whole cell extract from HeLa cells is loaded as positive control. HeLa cells were lysed using the Bioruptor ®. PROTEIN EXTRACTION KIT PAGE 10 DIAGENODE M A N U A LInnovating Epigenetic SolutionsPAGE 11 |PROTEIN EXTRACTION KIT DIAGENODE 030812。

少花蒺藜草荧光定量PCR内参基因筛选与验证徐礼莹;田迅;百灵;吴佳丽;张福霖;黄菊;马文静;吕广超;马红悦;陈宇杰【期刊名称】《现代农业研究》【年(卷),期】2024(30)3【摘要】少花蒺藜草(Cenchrus pauciflorus Benth.)为禾本科蒺藜草属,是入侵我国北方地区的一年生恶性杂草,少花蒺藜草的入侵态势逐年扩增,因此对于少花蒺藜草的基因研究,以及在分子水平对于防治少花蒺藜草入侵的研究显得尤为重要。

本研究以少花蒺藜草转录组数据为基础,候选了6个内参基因,荧光定量PCR检测候选内参基因的表达,通过geNorm、NormFinder、BestKeeper、比较△Ct法和RefFinder五种评价方式,综合评估筛选了最佳内参基因。

结果表明,6个候选内参基因的综合排名为TCTP1>RH37>ACX4>TCTP2>GAPDH>TBP,TCTP1基因在少花蒺藜草不同时期不同组织样本中表达最高最稳定,最不稳定表达的是基因TBP,TCTP1为综合评估下少花蒺藜草的最佳内参基因,因此TCTP1可作为内参基因用于少花蒺藜草荧光定量PCR检测研究中。

本研究为少花蒺藜草后续的基因研究以及在分子水平上为少花蒺藜草生物防治提供一定的理论基础。

【总页数】7页(P10-16)【作者】徐礼莹;田迅;百灵;吴佳丽;张福霖;黄菊;马文静;吕广超;马红悦;陈宇杰【作者单位】内蒙古民族大学生命科学与食品学院【正文语种】中文【中图分类】S451【相关文献】1.茉莉花实时荧光定量PCR内参基因的筛选与验证2.青海湖裸鲤实时荧光定量PCR内参基因筛选与验证3.栝楼实时荧光定量PCR内参基因的筛选与验证4.川赤芍实时荧光定量PCR内参基因的筛选与验证5.阳桃(Averrhoa carambola)实时荧光定量PCR内参基因的筛选与验证因版权原因，仅展示原文概要，查看原文内容请购买。

·药物与临床·多黏菌素B 血药浓度的测定及其在重症患者中的应用Δ甘雨＊，喻明洁，刘芳，程林，陈勇川 #（陆军军医大学第一附属医院药学部，重庆　400038）中图分类号 R 978 文献标志码 A 文章编号 1001-0408（2023）06-0704-06DOI 10.6039/j.issn.1001-0408.2023.06.12摘要 目的 建立测定多黏菌素B 血药浓度的方法并应用于临床。

方法 血浆样品经5%三氯乙酸溶液蛋白沉淀后，以多黏菌素E 2为内标，采用超高效液相色谱-串联质谱（UPLC-MS/MS ）法测定多黏菌素B 1、B 2的质量浓度。

以BEH C 18为色谱柱，以水（含0.1%甲酸）-乙腈（含0.1%甲酸）为流动相进行梯度洗脱，流速为0.5 mL/min ，进样量为10 μL 。

采用电喷雾离子源以多反应监测模式进行正离子扫描，用于定量分析的离子对分别为m /z 603.2→101.2（多黏菌素B 1）、595.7→101.1（多黏菌素B 2）、578.5→101.1（内标）。

采用上述方法测定79例重症患者体内多黏菌素B 的血药浓度，记录患者急性肾损伤（AKI ）的发生情况并分析多黏菌素B 血药浓度与AKI 发生的相关性。

结果 多黏菌素B 1、B 2检测质量浓度的线性范围分别为200～20 000、50～5 000 ng/mL （r ＞0.995），定量下限分别为200、50 ng/mL ；日内、日间精密度的RSD 均不高于12.06%，平均提取回收率为103.04%～117.44%（RSD ≤10.45%），基质效应、稳定性试验的RSD 均不高于7.42%。

79例患者的多黏菌素B 稳态谷、峰浓度分别为（2.54±2.52）、（8.17±5.20）mg/L 。

在被纳入AKI 评价的27例患者中，有18例患者（66.67%）发生AKI ；未发生AKI 患者的多黏菌素B 峰浓度显著低于AKI 患者（P ＜0.05），但两者谷浓度比较差异无统计学意义（P ＞0.05）。

细胞自噬转录组+蛋白组
细胞自噬（autophagy）是一种细胞内的重要代谢过程，通
过分解和回收细胞内的蛋白质、脂质和其他细胞器，维持
细胞内环境的稳定性。

细胞自噬包括三个主要步骤：识别
和包裹、溶酶体融合和降解。

在细胞自噬的过程中，转录组（transcriptome）和蛋白组（proteome）起着重要的作用。

转录组是指细胞中所有基
因的转录产物，即RNA的总体表达情况。

蛋白组是指细胞
中所有蛋白质的总体表达情况。

细胞自噬的转录组研究主要关注细胞自噬相关基因的表达
变化。

通过比较自噬诱导条件下和正常条件下的转录组数据，可以发现与细胞自噬相关的基因的表达差异。

这些基
因包括自噬相关基因（如ATG基因家族）、信号通路调控
基因、膜蛋白基因等。

转录组研究可以帮助我们了解细胞
自噬的调控机制以及自噬在不同生理和病理状态下的变化。

细胞自噬的蛋白组研究主要关注细胞自噬相关蛋白的表达
和修饰变化。

通过质谱分析等技术，可以鉴定和定量自噬
相关蛋白的表达水平和修饰状态，如磷酸化、乙酰化、泛
素化等。

这些蛋白包括自噬相关蛋白（如LC3、Beclin-1）、信号通路调控蛋白、膜蛋白等。

蛋白组研究可以帮助我们
了解细胞自噬的分子机制以及自噬在不同生理和病理状态
下的变化。

综上所述，细胞自噬的转录组和蛋白组研究可以帮助我们
深入了解细胞自噬的调控机制和分子机制，为相关疾病的治疗和药物开发提供重要的理论基础。

黄芩提取物中黄酮类成分血浆蛋白结合率的测定冯志强;韩坚;谢智勇;廖琼峰;张蕾【期刊名称】《中国药理学通报》【年(卷),期】2012(028)002【摘要】Aim To establish a RP-HPLC method for determination of concentration of four major flavonoids, namely baicalin ( BL ), wogonoside ( WL ), wogonin ( W ) and oroxylin A ( OA ), in the total flavonoids extract of scutellaria baicalensis Georgi in rat plasma and to study the plasma protein binding rate. Methods The equilibrium dialysis was carried out to determine the plasma protein binding rate of the target compounds. The plasma samples were extracted by protein precipitation with methanol. With the use of maackiain as the internal standard, the concentration of the multicomponent inside and outside the dialysismembranes was determined by HPLC method. Results The plasma protein binding rates of baicalin and wogonoside decreased with increasing of concentration in a concentration-dependent manner( BL: 82.03% →77.25% →67. 17% ; WL: 82. 24%→ 76. 08% →69. 87% ), while were not the same of wogonin and oroxylin A. The average plasma protein bingding rates of wogonin and oroxylin A were ( 89. 66 ± 1. 19 )% and ( 88. 56 ± 1. 87 )% , respectively. Conclusions The HPLC method established in the paper is simple, sensitive, little interfe-rential and stable. The four major compounds in the total fla-vonoids extract of scutellaria baicalensis Georgi have high capacityin binding plasma protein in rat in vitro.%目的建立同时测定黄芩总黄酮部位中4 种黄酮类成分在大鼠血浆中药物浓度的方法,并测定其体外血浆蛋白结合率.方法平衡透析法测定黄芩黄酮类成分血浆蛋白结合率,以高丽槐素为内标,生物样本用甲醇沉淀蛋白进行预处理,采用HPLC法测定4种黄酮类成分在透析内、外液中的浓度.结果在低、中、高3 种浓度下,黄芩苷(BL)和汉黄芩苷(WL)的血浆蛋白质结合率随给药浓度的增大而降低(BL:82.03%→77.25%→67.17%;WL:82.24%→76.08%→69.87%),而汉黄芩素(W)和千层纸素A(OA)则呈现非质量浓度依赖性,其平均血浆蛋白结合率分别为(89.66±1.19)%、(88.56±1.87)%.结论该文所建立的测定方法简便、灵敏、专属性强.4个黄酮类成分在大鼠体外均有较高的血浆蛋白结合率.【总页数】4页(P286-289)【作者】冯志强;韩坚;谢智勇;廖琼峰;张蕾【作者单位】广州中医药大学中药学院,广东,广州,510006;中山大学药学院,广东,广州,510080;广州中医药大学中药学院,广东,广州,510006;中山大学药学院,广东,广州,510080;广州中医药大学中药学院,广东,广州,510006;广州中医药大学中药学院,广东,广州,510006【正文语种】中文【中图分类】R-332;R282.71;R284.1;R341;R969.1【相关文献】1.超高效液相色谱-串联质谱法同时测定黄芩提取物中4种黄酮类成分 [J], 段云2.HPLC法测定枳芩颗粒中黄酮类成分的含量 [J], 高红宁;潘雨柔;殷奕;毛春芹;陆兔林3.HPLC法同时测定三角草中黄酮类成分异荭草苷和异牡荆素的含量 [J], 陈冠;王蕾;王宏;江茜;刘琳娜4.高效液相色谱法测定金银花中黄酮类成分槲皮素的血浆蛋白结合率 [J], 修玮5.翻白草中黄酮类成分含量测定及其质量评价研究 [J], 孔晓妮;吕华瑛;周洪雷因版权原因，仅展示原文概要，查看原文内容请购买。

桑黄提取物体内抗肿瘤作用的实验研究桑黄（Sophora flavescens）是一种传统中药，被广泛应用于中医临床实践中治疗多种疾病，包括肿瘤。

许多研究表明，桑黄提取物具有抗肿瘤的活性。

本实验旨在探究桑黄提取物在小鼠模型中的抗肿瘤作用。

我们从中药市场购买了粉碎后的桑黄根和根茎，并对其进行颗粒化处理。

接下来，我们使用乙醇对桑黄颗粒进行提取，得到了桑黄乙醇提取物。

为了评价桑黄提取物的抗肿瘤作用，我们采用了腹腔注射肿瘤细胞法，将小鼠随机分为实验组和对照组。

实验组小鼠腹腔注射了肿瘤细胞后，开始接受桑黄提取物的处理。

对照组则接受了相同的操作，但没有给予桑黄提取物处理。

通过监测实验组和对照组小鼠的生存时间和肿瘤大小，我们发现实验组小鼠的生存时间明显延长，肿瘤大小也明显减小，与对照组相比具有显著差异。

这表明桑黄提取物具有抗肿瘤的活性。

为了进一步研究桑黄提取物的抗肿瘤作用机制，我们进行了组织切片和免疫组化分析。

结果显示，在实验组中，肿瘤组织中的凋亡标志物（如活化的半胱氨酸蛋白酶-3）的表达明显增加，而增殖标志物（如Ki-67）的表达明显下降。

这表明桑黄提取物通过促进肿瘤细胞凋亡和抑制细胞增殖来发挥抗肿瘤作用。

我们还检测了实验组和对照组小鼠的免疫功能。

结果显示，实验组小鼠的免疫功能显著增强，包括NK细胞活性、淋巴细胞亚群比例和细胞因子水平。

这表明桑黄提取物通过增强机体免疫功能，加强对肿瘤的免疫监视来发挥抗肿瘤作用。

本实验证明桑黄提取物具有抗肿瘤作用，可能通过促进肿瘤细胞凋亡、抑制细胞增殖和增强机体免疫功能来发挥其作用。

这为进一步研究桑黄提取物的抗肿瘤机制和开发新的抗肿瘤药物奠定了基础。

大蓟提取物改善高胆固醇血症模型小鼠的代谢组学研究Δ高梦梦 1＊，陈桢琳 1，郝雅坤 2，郭姣 1 #（1.广东药科大学中医药研究院/广东省代谢病中西医结合研究中心/糖脂代谢病教育部重点实验室/广东省代谢性疾病中医药防治重点实验室，广州　510006；2.广东药科大学中药学院，广州　510006）中图分类号 R 285.5 文献标志码 A 文章编号 1001-0408（2023）13-1590-06DOI 10.6039/j.issn.1001-0408.2023.13.09摘要 目的 基于代谢组学技术探究大蓟提取物改善高胆固醇血症的作用机制。

方法 以大孔树脂吸附法制备大蓟提取物，并利用液相色谱-质谱联用仪鉴定其主要成分。

实验小鼠先随机分为对照组（n ＝6）和造模组（n ＝16），造模组小鼠采用饮食诱导建立高胆固醇血症模型，造模成功后，再将造模组小鼠分为模型组（n ＝8）及大蓟提取物组（n ＝8）。

大蓟提取物组小鼠灌胃大蓟提取物400 mg/（kg ·d ）（以提取物计），其余2组小鼠灌胃等体积0.3%羧甲基纤维素钠溶液，持续6周。

给药结束后，以血清总胆固醇（TC ）、甘油三酯（TG ）水平和肝脏组织病理变化评价大蓟提取物的干预效果，并通过代谢组学方法探讨大蓟提取物改善高胆固醇血症模型小鼠的相关机制。

结果 从大蓟提取物中共鉴定出绿原酸、蒙花苷、柳穿鱼叶苷等12种成分。

给药6周后，与对照组比较，模型组小鼠血清中TC 水平显著升高、TG 水平显著降低（P ＜0.05），肝脏组织出现大量脂滴，肝细胞排列紊乱，肝索结构被破坏。

与模型组比较，大蓟提取物组小鼠血清中TC 水平显著下降（P ＜0.05）；肝脏组织中的脂滴明显减少，肝细胞以中央静脉为中心呈放射状紧密排列，肝索排列整齐。

代谢组学研究显示，大蓟提取物干预后，乙醇胺、富马酸、胆固醇等代谢产物水平发生显著回调；最终得到丙氨酸-天冬氨酸-谷氨酸代谢、精氨酸生物合成、柠檬酸循环3条代谢通路。