怪不得师姐说这是毫无积累（没有学术资源）的课题组呢

不想看文献

我觉得我总是在某个步骤上卡很久

比如现在，我应该要画一个这篇文献的流程图，但是事实上我还没搞清楚这篇文献各个实验都为了验证什么（逻辑很劝退）

（主要是我真的没法理解逻辑，也很怀疑根本就没有逻辑，我也不是很想要搞这个选题（还没有师姐的选题有搞头

但是如果要搞师姐的选题就要整理一大堆看不懂的数据（从里面筛选出要搞的分子）

而且最卡关的是我在两种思路中摇摆不定

炎症因子究竟是伤害性还是保护性还是两者都有

看了几天这篇文献我还是没搞清楚
A到C B到C都看懂了
但A到B究竟有没有逻辑啊(究竟有没有直接证明啊啊啊啊啊啊啊)

这种思路其实是挺垃圾的
我真的不想搞这个

又到了传统想死之看文献环节

肿瘤的文章就是好发啊

这篇回顾比上篇好一点

这篇比上篇好读很多，不知道是不是因为上一篇作者英语不好，句子都很绕

（到现在看的全都是中国发的论文，也没办法）

primary culture 原代培养

formalin-fixed paraffin embedded
福尔马林固定石蜡包埋

看了一会儿就昏昏欲睡，下午要搞执医报名然后运动一下就要去吃饭了

玩嗨了，差点都忘了明天要和师姐去实验室跑电泳
好emo，我自己的课题还是没有着落，很快就花粉季了，又一直拖延不看文献，导师对科研一窍不通

心态belike：人生不在工作，临床不在科研

摆大烂

已经一个月没碰课题嘟串了

FSTL1-knockdown improves neural oscillation via decreasing neuronal-inflammation regulating apoptosis in Aβ1–42 induced AD model mice
好烦啊，学校账号也没买这篇文献
scihub和librarygenesis都没有搜到

要学学怎么用AI hack吗
师姐在一个付费群里面，那个群用AI hack出来赚中间价

To determine if a long noncoding RNA (lncRNA) has cis-regulatory functions, several criteria are typically considered:

Proximity to Target Gene: The lncRNA should be located near the target gene on the same chromosome.
Expression Correlation: There should be a correlation between the expression of the lncRNA and the target gene.
Functional Evidence: Experimental evidence, such as loss-of-function or gain-of-function studies, should show that the lncRNA can regulate the target gene’s expression.
Physical Interaction: The lncRNA should physically interact with the target gene’s DNA or associated chromatin.
Conservation: The regulatory function of the lncRNA should be conserved across different species or cell types, if applicable.

瞅了瞅这234不就是师姐的主要的实验嘛
表达相关性、敲低之后再看、再来个荧光素酶之类的确定是有结合的

Testing the proximity of a long noncoding RNA (lncRNA) to its target gene involves several methods:

Genomic Location Analysis: By examining the genomic coordinates of lncRNAs and potential target genes, researchers can determine physical proximity within the genome.
Expression Studies: Correlating the expression levels of lncRNAs and nearby genes can suggest regulatory relationships, especially when changes in lncRNA expression affect the gene’s expression.
Chromatin Conformation Capture (3C): Techniques like 3C or Hi-C can be used to detect physical interactions between the lncRNA and the target gene’s DNA, indicating proximity within the three-dimensional structure of the genome.
Computational Prediction: Databases like LncRNA2Target provide information on experimentally validated lncRNA-target interactions, which can be used to predict potential cis-regulatory relationships.
Experimental Validation: Techniques such as RNA immunoprecipitation or RNA pull-down assays can confirm the physical interaction between the lncRNA and the target gene’s DNA or RNA1.
These methods collectively help in establishing the cis-regulatory role of lncRNAs by confirming their proximity to target genes.

what the whole shitty thing is exactly?

ceRNA stands for competing endogenous RNA. In molecular biology, ceRNAs are transcripts that regulate other RNA molecules by competing for shared microRNAs (miRNAs). This interaction is part of a complex regulatory network where ceRNAs can influence the levels and activity of miRNAs, thereby affecting the expression of miRNA targets.

The ceRNA network includes various types of RNAs such as long noncoding RNAs (lncRNAs), pseudogenes, and circular RNAs (circRNAs), all of which can act as molecular sponges. They bind to miRNAs through miRNA response elements (MREs), reducing the miRNAs’ ability to target other RNAs. This mechanism is crucial for maintaining the balance of gene expression and has implications in various biological processes and diseases, including cancer

https://www.lncrnablog.com/lncrnatargets-a-platform-for-lncrna-target-prediction-based-on-nucleic-acid-thermodynamics/

[摘自公众号文章]
因此这种模式来预测靶基因的原理就是预测lncRNA序列与mRNA序列结合的自由能，自由能越小，越容易结合。

说来变态反应的炎症和一些其他炎症的分子机制一样吗？

FSTL1 flox/+ 和 FSTL1 +/- 小鼠之间的区别在于它们的基因构成和由此产生的生理效应：

FSTL1 flox/+ 小鼠：这个称呼通常指的是小鼠中有一个FSTL1基因被loxP位点包围（flox），这允许在特定组织或特定时间通过Cre重组酶的结合来有条件地删除该基因。"+"表示存在一个正常的、非flox的FSTL1基因的等位基因。
FSTL1 +/- 小鼠：这些是杂合子小鼠，其中一个FSTL1基因被敲除（因此是"-“），另一个是正常的基因（因此是”+"）。这些小鼠可以用来研究FSTL1基因表达减少的效果。

文献读不懂的地方就应该直接问AI

相比之下，除非使用Cre重组酶特异性地删除flox的FSTL1基因，否则FSTL1 flox/+ 小鼠不会表现出这些效应。提到的研究没有直接提供未经Cre介导的重组的FSTL1 flox/+ 小鼠的信息，但暗示如果没有基因删除，它们不会表现出与FSTL1 +/- 小鼠相同的抵抗力1。

神td暗示，怪不得我读不懂

The flox and Cre systems are integral parts of genetic engineering, particularly in creating genetically modified organisms like mice for research purposes. Here’s a more detailed explanation:

Flox: The term “flox” refers to a DNA sequence that has been flanked by loxP sites. These loxP sites are specific 34-base pair sequences that are recognized by the Cre recombinase enzyme1. When a gene is “floxed,” it means that it can be conditionally removed or altered by the action of Cre recombinase, which allows researchers to study the function of the gene in a controlled manner.
Cre: Cre is an enzyme called Cre recombinase that comes from bacteriophage P1. It recognizes loxP sites in the DNA and can cut and rejoin the DNA at these sites1. This allows for precise manipulation of the genome, such as the deletion, insertion, or inversion of DNA segments. The activity of Cre recombinase can be controlled so that it only acts in specific cell types or in response to certain stimuli, making it a powerful tool for studying gene function and creating models of human disease2.
The Cre-Lox system is widely used in neuroscience and other fields of biology to study complex systems and diseases. For example, it can be used to delete a gene in only certain types of neurons, which helps scientists understand the role of that gene in brain function1. It’s also used in creating models for diseases like cancer, where researchers can activate or deactivate cancer-related genes in specific tissues1.

This system provides a high level of control over gene expression and is invaluable for research that requires the study of gene function in living organisms. It’s one of the most precise tools available for genetic manipulation and has revolutionized the field of genetics.

感觉这里的小破实验室应该不具备这项技术

1. 定义研究目标
- 探索 circ_0002111 在甲状腺乳头状癌（PTC）进展中的调控机制

2. 提出假设
- circ_0002111 在 PTC 中上调并影响细胞增殖、迁移和糖酵解

3. 设计实验
a. RT-qPCR 测量 circ_0002111 表达水平
b. MTT 实验检测细胞增殖
c. EdU 掺入实验检测 DNA 合成
d. 集落形成实验检测长期增殖
e. Transwell 实验检测细胞迁移
f. Seahorse 分析进行代谢剖析
g. 西方印迹法分析蛋白表达
h. 小鼠体内移植瘤实验检测体内肿瘤形成能力
i. RNA 拉下实验确定 circ_0002111 的相互作用
j. 双荧光素酶报告基因表达调控实验

4. 进行实验
- 执行上述实验并记录观察结果

5. 数据分析
- 与对照组比较结果，确定 circ_0002111 的效果

6. 得出结论
- 评估数据是否支持假设

7. 报告发现
- circ_0002111 通过海绵 miR-134-5p 和调节 FSTL1 促进 PTC 进展

Here’s a summary of the key experiments and their conclusions from the article:

Circ_0002111 Expression Analysis: The study found that circ_0002111 was significantly upregulated in PTC tissues and cells, suggesting its role in PTC progression1.
Functional Assays: Silencing circ_0002111 inhibited PTC cell proliferation, migration, and glycolytic metabolism, indicating its oncogenic function23.
In Vivo Tumorigenesis: Knockdown of circ_0002111 reduced tumor growth in a xenograft model, further confirming its role in promoting PTC4.
miR-134-5p Interaction: Circ_0002111 was shown to act as a sponge for miR-134-5p, affecting its expression and PTC cell behavior.
FSTL1 Regulation: The study demonstrated that miR-134-5p targets FSTL1, and circ_0002111 can regulate FSTL1 levels by sponging miR-134-5p.

反转录定量聚合酶链反应 (RT-qPCR)
用于检测circ_0002111、miR-134-5p和FSTL1的水平。1
通过SYBR™ Green Master Mix和TaqMan™ Fast Advanced Master Mix进行定量反应。
细胞增殖评估
使用MTT试剂、EdU试剂和克隆形成试验来评估细胞增殖。
通过显微镜观察和图像分析软件计数。
细胞迁移能力测定
通过Transwell试验来确定细胞迁移能力。2
使用结晶紫染色并通过倒置显微镜观察。
糖酵解分析
通过外部酸化率(ECAR)、氧消耗率(OCR)、葡萄糖消耗和乳酸产生来分析糖酵解。
使用Seahorse XFe 96 Extracellular Flux Analyzer进行检测。

1. Define Research Objective
- Explore regulatory mechanism of circ_0002111 in PTC progression

2. Formulate Hypothesis
- circ_0002111 upregulates in PTC and affects cell proliferation, migration, and glycolysis

3. Design Experiments
a. RT-qPCR to measure circ_0002111 expression levels
b. MTT assay for cell proliferation
c. EdU incorporation assay for DNA synthesis
d. Colony formation assay for long-term proliferation
e. Transwell assay for cell migration
f. Seahorse analysis for metabolic profiling
g. Western blot for protein expression analysis
h. Xenograft tumor assay in mice for in vivo tumorigenicity
i. RNA pull-down to identify circ_0002111 interactions
j. Dual-luciferase reporter assay for gene expression regulation

4. Conduct Experiments
- Perform the above assays and record observations

5. Analyze Data
- Compare results with control groups to determine the effect of circ_0002111

6. Draw Conclusions
- Assess whether the data supports the hypothesis

7. Report Findings
- circ_0002111 promotes PTC progression by sponging miR-134-5p and regulating FSTL1