Chapter 66: Huge Gains (Please Collect, Please Follow)

The National High-Tech Research and Development Program (863 Program) and the National Key Basic Research Development Program (973 Program, including Major Scientific Research Programs) are both relatively super-high-end national research projects.

But anyone who wants to obtain one of them needs sufficient accumulation.

Of course, anyone who can obtain one can also achieve fame and success; several million yuan in project funding is a piece of cake, and tens of millions of yuan in research funds are also very common.

And the startup funding for such a project is more than a hundred times the Youth Fund Yuan Weihong currently obtained.

When he was about to say something else, the patient who had finished the examination and follow-up consultation outside had already pushed open the door.

Yuan Weihong stopped speaking, and Fang Ziye also hurriedly began to harvest his Knowledge Points, reviewing films step by step, making precise diagnoses and prescribing medication, and then obtained 0.9 Knowledge Points.

He happily became smug.

Because even at Zhongnan Hospital, it was not a place where difficult and complex cases were everywhere; although there were naturally not a few difficult and complex cases, there were also simple disease types, and patients would not consider such a statement as wasting resources when registering.

After the outpatient clinic ended, Fang Ziye obtained about 15 Knowledge Points again; with this, even after adding points to raise Knot Tying Technique to Level 3 and giving gifts to the teacher,

the balance of Knowledge Points suddenly accumulated to as many as 60, allowing him to add a Level 3 skill at the same time; tonight, Fang Ziye also had a cell experiment that was about to yield results; when he observed the results, it would definitely be another considerable amount of Knowledge Points.

"Master, I'll put away my white coat first, then go to the laboratory!"

"Okay, wb run well." Yuan Weihong instructed, as if hinting to Fang Ziye that he knew exactly what Fang Ziye was doing.

Fang Ziye nodded and quickened his pace.

Doing experiments, making money, publishing papers, gaining Knowledge Points—fulfilling and happy.

wb is an abbreviation; Protein Immunoblotting (westernblot) is a method of transferring proteins onto a membrane and then using antibodies for detection.

The ultimate result of gene expression is the production of corresponding proteins (or enzymes). The hk2 Fang Ziye studies is actually Hexokinase 2, the first rate-limiting enzyme in the biochemical glycolysis process; hk2 is the abbreviation for Hexokinase 2, which is an enzyme that catalyzes the phosphorylation of hexose; it is the first enzyme in the glycolysis pathway and also the absolute rate-limiting enzyme of the glycolysis pathway.

It is well known that glycolysis is the first step of sugar conversion into energy; without the first step of glycolysis, it is almost impossible for glucose and fructose to be normally utilized, and if relying on ketone bodies and other bypasses as substrates for aerobic oxidation, it will definitely be of no avail.

Therefore, quantitatively detecting the content of hk2, the absolute quantity after overexpression and silence of Giant Cell Tumor of Bone, is quantitative evidence for detecting the role of hk2 in the development of Giant Cell Tumor of Bone.

westernblot uses Polyacrylamide Gel Electrophoresis; the detected object is protein, the "probe" is antibody, and "color development" uses labeled Secondary Antibody.

Protein samples separated by page are transferred onto a solid-phase carrier (such as Nitrocellulose Membrane); the solid-phase carrier adsorbs proteins via non-covalent bonds and can maintain the polypeptide types and biological activity after electrophoresis unchanged. Using the protein or polypeptide on the solid-phase carrier as an antigen, it undergoes an immune reaction with the corresponding antibody, then reacts with a second antibody labeled with enzyme or isotope; after Substrate Color Development or Radioautography, the protein components of specific target gene expression separated by electrophoresis can be detected...

The principle sounds quite complex, but actually when truly operating, the main steps are mainly the following few steps.

Reagent Preparation, which is the runway for proteins to run, needs to be provided by Polyacrylamide Gel Electrophoresis.

Then comes the competing athletes, which is lysing the Tumor Cells so that internal proteins are fully released, and then making a Standard Curve.

After making the Standard Curve, the 'athletes' produced by the cells he processed can start running on the track.

Immediately after electrophoresis, transfer, immune reaction, development, and other series of steps, through special antibodies and reagents, the film of the target protein can be displayed.

Scan or photograph the film, and use a gel image processing system to analyze the molecular weight and net optical density value of the target band...

This process is very complex, relatively long, and requires more patience.

However, Fang Ziye had already finished the outpatient clinic anyway; after coming to the laboratory, he did it step by step according to the rules. And beside him, Luo Tingzhu helped with small favors.

After the results were finally presented, Luo Tingzhu was overjoyed: "Senior Brother Fang, your band today shows very clearly; last time what Senior Brother Fan ran out was Dumbbell-Shaped Band."

After speaking, she tilted her head to look at Fang Ziye's film; the band was clear, with no杂 bands, and the background was clean!

"A Dumbbell-Shaped Band generally means there is a problem with the gel preparation; another possibility is that the sample contains too many impurities that were not centrifuged down, and the impurities deposit in the middle of the well, so the protein is naturally squeezed to both sides."

"After preparing the gel, unqualified ones must not be used; do not take risks." Fang Ziye thought about the principle carefully and explained it to Luo Tingzhu like this.

"What if there is Band Tailing?" Luo Tingzhu thought for a moment and asked another question; it should be something she had seen when following others to do experiments.

"If there is Band Tailing, it is generally because the protein concentration is high, or the Primary Antibody concentration and time are too long."

"If the protein content is high, it may be that the literature preparation in the early stage was not determined; I think the most likely reason is that the Primary Antibody concentration is too high and the action time is too long."

"Of course, there are other reasons." Fang Ziye looked at his film again.

hk2 and b-actin (Actin) Internal Reference two bands, with a total of three groups: Overexpression, Normal, and Silence.

The Internal Reference of Tumor Cell experiments, especially cell experiment Internal References involving Glycolysis Pathway, cannot use common gapdh (Glyceraldehyde-3-Phosphate Dehydrogenase).

"This graph of yours can be directly used for plotting with prism, right?" Luo Tingzhu continued to ask.

"Should be about the same; the thickness and grayscale difference of the bands is still quite obvious. Take a photo, take a photo..." Fang Ziye hurriedly asked Luo Tingzhu to help take a photo too.

Luo Tingzhu agreed readily, lively like a diligent little squirrel.

And Fang Ziye at this moment, watching Knowledge Points jump rapidly to 134, suddenly increased by a full seventy Knowledge Points, also knew that the role of this wb experiment in this research project was not small; it might be the small skeleton supporting the scale of the entire project.

"How is this photo taken?" After taking the photo, Luo Tingzhu asked Fang Ziye.

Fang Ziye showed a smug smile, while Luo Tingzhu thought her photography level was not bad, so she took several more photos with click-click sounds.

Then it was cleaning the laboratory bench and washing the corresponding Experimental Equipment.

After everything ended, Fang Ziye suggested: "Junior Sister Luo, you worked hard; let's go eat a Late-night Snack together?"

Luo Tingzhu was currently flipping through the photos of the wb results, tilting her head and clicking/selecting: "Okay."

(Chapter End)