The article describes a series of experiments to investigate the role of Follistatin-like protein 1 (FSTL1) in asthma. Here’s a summary of the methods and their intended purposes, along with the results:

OVA-Induced Asthma Model: Mice were sensitized and challenged with ovalbumin (OVA) to induce asthma-like symptoms. The control groups received PBS. This model was used to assess the role of FSTL1 in asthma.
Result: Fstl1± mice showed reduced Muc-5AC production and mucus hypersecretion compared to OVA-WT mice1.
Immunohistochemistry & ELISA: These methods were used to detect the protein levels of Muc-5AC, FSTL1, NLRP3, and IL-1β in lung tissues and bronchoalveolar lavage fluid (BALF)2.
Result: Fstl1± mice had decreased levels of these proteins, indicating reduced inflammation.
Western Blot Analysis: This technique was employed to measure the protein expression of NLRP3 and IL-1β in alveolar macrophage cells (MH-S cells)3.
Result: Pretreatment with siFSTL1 or MCC950 reduced the production of NLRP3 and IL-1β induced by OVA or FSTL14.
Cell Culture Experiments: MH-S cells were stimulated with FSTL1 and/or OVA to explore the effect of FSTL1 on the NLRP3/IL-1β signaling pathway.
Result: FSTL1 stimulation increased the expression of NLRP3 and IL-1β, while MCC950 pretreatment inhibited this effect56.
Overall, the study demonstrated that FSTL1 plays a significant role in allergic airway inflammation by activating the NLRP3/IL-1β signaling pathway, and that inhibiting FSTL1 could be a potential therapeutic strategy against asthma78. The experiments collectively aimed to elucidate the mechanisms of FSTL1 in asthma and to explore potential treatments by modulating this protein’s activity.

只能说现在做PPT美观确实很容易，AI确实可以的

The article explains various assays used to assess cell proliferation in the context of papillary thyroid carcinoma (PTC) research:

MTT Assay: This assay measures cell proliferation by adding MTT solution to cells1. MTT is reduced by metabolically active cells, producing a colored formazan product. The intensity of the color, measured by absorbance at 490 nm, correlates with the number of viable cells.
EdU Assay: EdU (5-ethynyl-2’-deoxyuridine) is a nucleoside analog of thymidine and is incorporated into DNA during active DNA synthesis. After labeling cells with EdU, a fluorescent dye is used to detect the incorporated EdU, allowing visualization and quantification of proliferating cells.
Colony Formation Assay: This assay evaluates the ability of a single cell to grow into a colony. Cells are plated at low density and allowed to grow until visible colonies form. The colonies are then fixed, stained, and counted to assess the proliferative potential of the cells.
These assays are utilized in the study to investigate the role of circ_0002111 in PTC cell proliferation and its potential as a therapeutic target2.

Linear PSD3 refers to the linear form of the gene Pleckstrin And Sec7 Domain Containing 3 (PSD3). In the context of this article, PSD3 is the parent gene from which circ_0002111, a circular RNA (circRNA), is derived. The article explains that circ_0002111 is produced from exons 5-9 of the PSD3 gene and has a mature length of 538 base pairs1. Unlike linear RNAs like PSD3, circRNAs like circ_0002111 form a covalently closed continuous loop, which makes them resistant to degradation by RNA exonucleases. This stability allows circRNAs to have regulatory roles in various cellular processes, including acting as microRNA (miRNA) sponges, influencing gene expression, and potentially contributing to diseases such as cancer2. In the case of papillary thyroid carcinoma (PTC), the article discusses the role of circ_0002111 in promoting malignant behaviors by regulating the miR-134-5p/FSTL1 molecular network34. The study uses RT-qPCR to show that linear PSD3 is significantly reduced after RNase R treatment, while circ_0002111 remains stable, highlighting the distinct properties of circRNAs compared to their linear counterparts5.

The Transwell assay, according to this article, is a method used to assess the migration ability of papillary thyroid carcinoma (PTC) cells. Here’s a brief overview:

Purpose: It’s utilized to evaluate the cell migration capability.
Procedure: Cells are placed in the upper chamber of a Transwell chamber, with cell culture medium in the lower chamber. After incubation, migrated cells are stained and counted.
Significance: The assay helps determine the impact of circ_0002111 on PTC cell migration, showing that circ_0002111 knockdown suppresses migration1.
Results Interpretation: Reduced cell migration in the assay indicates the effectiveness of circ_0002111 silencing in inhibiting PTC cell migration.
This assay is part of the experimental methods used to explore the regulatory mechanism of circ_0002111 in PTC progression

incubation
the process of incubating eggs, cells, bacteria, a disease, etc.

In biology, “knockdown” refers to an experimental technique used to reduce the expression of one or more genes in an organism. This reduction in gene expression can be achieved through various methods, such as the use of small interfering RNA (siRNA) or short hairpin RNA (shRNA), which bind to mRNA transcripts and prevent them from being translated into proteins. The term can also refer to the genetic modification that leads to a decrease in gene expression without completely eliminating it, which is different from a “knockout” where the gene is completely inactivated1.

Knockdown is often used to study the function of a particular gene by observing the effects of decreased gene expression on the organism. It’s a valuable tool for researchers to understand gene function and to investigate the potential role of genes in diseases and disorders

In summary, knockdown reduces gene expression temporarily without altering the DNA, while knockout permanently removes or inactivates the gene at the DNA level.

AI联想问题正好是我想问的

Here are the key points about OCR and the compounds oligomycin, FCCP, and Rote/AA:

Oxygen Consumption Rate (OCR): OCR is a measure of mitochondrial function and cellular metabolism. It indicates the rate at which cells consume oxygen, which is essential for energy production through oxidative phosphorylation1.
Oligomycin: This compound is an inhibitor of ATP synthase, an enzyme involved in the synthesis of ATP during oxidative phosphorylation. It is used to assess mitochondrial function by blocking ATP production and measuring the resulting changes in OCR2.
FCCP (Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone): FCCP is a chemical that uncouples oxidative phosphorylation by transporting protons across the mitochondrial membrane, leading to increased oxygen consumption without ATP production. It is used to determine the maximum respiratory capacity of mitochondria2.
Rotenone/Antimycin A (Rote/AA): These are inhibitors of the mitochondrial electron transport chain. Rotenone inhibits complex I, while Antimycin A inhibits complex III. They are used to measure non-mitochondrial oxygen consumption by shutting down mitochondrial respiration

The Xenograft Tumor Assay in this article refers to an experimental procedure used to study the function of circ_0002111 in tumor growth in vivo13. Here’s a brief overview:

Purpose: To analyze the role of circ_0002111 in promoting tumorigenesis in papillary thyroid carcinoma (PTC).
Method: BALB/c male nude mice were injected subcutaneously with PTC cells that were either untreated or had circ_0002111 knocked down.
Observations: Tumor volume and weight were measured over 4 weeks, and the results showed that tumors in the group with circ_0002111 knockdown were significantly smaller.
Conclusion: The assay demonstrated that circ_0002111 facilitates tumor growth in PTC, indicating its oncogenic role in vivo2.
This experiment supports the overall finding that circ_0002111 acts as a tumorigenic factor in PTC by targeting the miR-134-5p/FSTL1 pathway

好想摆烂啊，有六篇文章但是我还在第一篇打转

The logic behind the statements from the article is as follows:

Circ_0002111 as miR-134-5p Sponge: Circ_0002111 acts as a “natural sponge” for miR-134-5p, meaning it binds to and sequesters miR-134-5p1. This prevents miR-134-5p from performing its normal functions, such as regulating gene expression.
Tumor Progression Inhibition: The si-circ_0002111#1 (a small interfering RNA designed to target circ_0002111) was shown to inhibit tumor progression. However, when miR-134-5p levels were reduced, this inhibition was reversed, indicating that miR-134-5p is essential for the tumor-suppressive effects of si-circ_0002111#1.
FSTL1 as Target Gene: FSTL1 is identified as a target gene for miR-134-5p23. This means that miR-134-5p can bind to the mRNA of FSTL1 and regulate its expression.
The axis confirmation refers to the experimental validation of the interactions between circ_0002111, miR-134-5p, and FSTL14. This was confirmed through various assays such as dual-luciferase reporter assays, RNA pull-down assays, and RT-qPCR, which demonstrated the direct binding and regulatory relationships among these molecules5. These interactions form a regulatory axis that influences the progression of papillary thyroid carcinoma (PTC). The study aimed to elucidate the specific functional mechanism of circ_0002111 in PTC by investigating this molecular network

I’m sorry, but the web page you are referring to seems to be empty or has failed to load, so I cannot provide an explanation based on that article. However, generally speaking, an RNA pull-down assay is a technique used in molecular biology to investigate the interaction between RNA and proteins. It involves binding RNA molecules to a substrate, then “pulling down” the RNA along with any proteins that are bound to it. This allows researchers to identify and study the proteins that interact with specific RNA sequences. If you have another source or article you’d like me to summarize or explain, please let me know!

RNA Pull-Down Assay:

Purpose: It’s a technique used to determine the interaction between RNA and other molecules, such as proteins or microRNAs (miRNAs).
Process: Involves using a probe (like circ_0002111) to capture target miRNAs from cell extracts, which are then identified and quantified.
Application: This study used RNA pull-down to confirm that circ_0002111 binds to miR-134-5p, indicating a regulatory interaction in papillary thyroid carcinoma (PTC).
Significance: Helps understand the molecular mechanisms in diseases like PTC by revealing how circRNAs like circ_0002111 can regulate miRNA levels and affect gene expression.

The article verifies that circS100A11 enhances M2a macrophage activation through several key experiments:1

Expression Analysis: circS100A11 was found to be dominantly expressed in monocytes and significantly upregulated in children with asthma23.
Functional Assays: Knockdown of circS100A11 in THP-1-derived human macrophages resulted in reduced expression of M2a macrophage marker genes, while overexpression led to increased marker gene expression.
Protein Interaction: circS100A11 was shown to promote the translation of its host gene, S100A11, by competitively binding to CAPRIN1, which decreases the suppression of CAPRIN1 upon S100A11 translation45.
In Vivo Model: A cockroach allergen extract (CRE)-induced murine asthma model with macrophage-specific overexpression of circS100A11 exhibited exacerbated lung inflammation, confirming the role of circS100A11 in vivo.

今天：
问老板购买质粒的事宜
去实验室和师姐学习养细胞
去买一辆小电驴
晚上看执医技能视频，刷儿科的题

养完细胞了，啥都没记住，回去上网学
下一步去买小电驴

问了老板购买质粒事宜，我发现我就是害怕等消息回复，所以会拖延找老板，这次拖延了整一天

克服了消费冲动，在1600和2200选了便宜的，电池里程足够用了

晚上还有哪些事情：
在网上看一看养细胞的流程细节
找一下实验室某个型号的离心机怎么配平
还有执医
明天要看论文，细化预实验实验步骤，列出每一步需要的试剂，发给实验室的老师核对哪些有哪些没有

Cultivating COS-7 cells is a meticulous process that requires careful attention to aseptic techniques to prevent contamination. Here’s a detailed guide to help you with the cultivation process:

COS-7 Cell Cultivation Instructions:
Prepare the Growth Medium: Use Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS).
Incubation: Maintain the cells at 37°C in a humidified atmosphere containing 5% CO2.
Subculturing:
Rinse cells with sterile 1x PBS to remove all traces of media containing FBS.
For a T75 flask, add 2 mL of Trypsin-EDTA and observe cell layer detachment using an inverted microscope. Detachment should occur within 3-15 minutes.
If cells do not detach, incubate the flask to increase enzymatic activity. Avoid agitation during this process.
After detachment, add 8 mL of complete medium to neutralize the trypsin.
Use the appropriate split ratio aliquot of the cell suspension to transfer to a new culture flask.
Confluency: Maintain culture at a confluency between 30-90%. The doubling time for COS-7 cells is approximately 35-48 hours.
Long-term Storage: For storing COS-7 cells long-term, use the complete growth medium with 5% (v/v) DMSO and store in the vapor phase of liquid nitrogen12.
Using an Alcohol Lamp for Aseptic Techniques:
The alcohol lamp is used for flame sterilization, which is a part of aseptic technique. Here are the key points when you should use an alcohol lamp:

Before Starting: Sterilize all tools and work surfaces.
Preparing Culture Medium: Properly sterilize and cool the medium before use.
Aseptic Transfer: Flame sterilize the neck of the culture vessel to ensure no contaminants enter.
Remember, the use of an alcohol lamp should be part of your routine to maintain a sterile environment. It’s typically used before any procedure that requires opening of culture vessels or transferring cultures to prevent microbial contamination345.

Keep in mind that while using an alcohol lamp, you should work in a well-ventilated area and keep all flammable materials away from the flame. Always practice good personal hygiene, such as washing hands before and after handling cultures, and wear appropriate protective equipment.

I hope this helps you with your cell culture practice! If you need more assistance or have other questions, feel free to ask.

今天todolist
帮老板审批我的项目采购(？)
把信息发给厂商客服
上次没查的离心机怎么配平
查询TPH-1细胞培养出现细胞聚集正常吗
最重要:查文献细化预实验方案

周四前还要做好主任教学查房的PPT

一个manual就几十页，还有三份manual，最苟的是从头扫到尾没有解决我实际问题的part

Yes, in the TX-200 swinging bucket rotor, all six peripheral holes are designed to be equivalent in terms of balancing. The rotor allows the buckets to swing out under centrifugal force, and when the machine is not running, the buckets can rotate freely around the central hole.

For proper balancing, it’s important to ensure that each opposing pair of holes is balanced with tubes of equal weight. This means that if you place a tube in one of the peripheral holes, you should place a tube of the same weight directly opposite to it in the corresponding peripheral hole on the other side of the central hole.

艹虽然AI这次真的表现得很差，但是我在对话中自己想出来了

欣赏一下ai的回答
总共四个版本

Place your 1ml tube in hole A1.
Place your 2ml tube in hole B1.
Add a balance tube with 1ml of water in hole A4 (opposite A1).
Add a balance tube with 2ml of water in hole B4 (opposite B1).
Now, you have 2ml on side A (1ml in A1 and 1ml in A4) and 4ml on side B (2ml in B1 and 2ml in B4). To balance the sides, you need to add more tubes:

Add a balance tube with 2ml of water in hole A2.
Add a balance tube with 1ml of water in hole B2.
With these additions, you will have 4ml on side A (1ml in A1, 1ml in A4, and 2ml in A2) and 5ml on side B (2ml in B1, 2ml in B4, and 1ml in B2). To fully balance it, add one more tube:

Add a balance tube with 1ml of water in hole A5.

我想出来了，但是问题在于没有那么多配平的管子，如果想要在一次中搞定一管1ml和一管2ml的配平，至少要再来4管水（1+3）

想到最后就是因为每个离心5min，分开了也不算特别久，如果再长或者管子再多就配平吧

我目前是这么想的，不知道对不对

A-side: B-side:
A1 [1ml] B1 [1ml]
A2 B2
A3 B3
A4 [1ml] B4 [1ml]
A5 B5
A6 B6
A中心孔 [2ml] B中心孔 [2ml]

1 2ml 各一管应该没啥问题（吧？）
但是如果现在有1 2 3ml各一管呢？

烦死了，AI不靠谱，manual不详细，师姐靠不上
没人带就是一个离心机配平都要研究上一小时
还是去搜索引擎或者小红书搜一搜吧

艹为什么3不行啊

去扒拉原网
图3为放置两只试管时，虽为2点平衡受力，但是在高速运行中，在旋转的共振点区间，会产生较大的不平衡共振点；