NovelsAlex Chapter 1426 Surprise
Chapter 1426 Surprise
Chapter 1426 Surprise
Einstein stayed in the animal room for a long time, so long that Fritz thought he was asleep when he came back from the cryogenic freezer.
“Mannstein?” Fritz stood in the doorway, holding a frost-covered styrofoam box. “Are you alright?”
Mainstein stood up, his knees clicking. He stretched his numb legs and took the styrofoam box from Fritz. "The entire M21 organization... is here?"
"It's all here: seventeen wax blocks, eight tubes of frozen tissue, and three unstained slides." Fritz opened the styrofoam box, revealing neatly stacked numbered embedding cassettes and centrifuge tubes. "What do you need me to do?"
“Reslice, re-stain, double-label with corticosteroids and NeuroD, plus DAPI staining of the nuclei. I need to see clearly what these cells actually are.”
Fritz didn't ask why; he had worked with Mainstein for years and was used to such sudden instructions. He simply nodded, took the styrofoam box from Mainstein, and turned to walk towards the slide room. After taking a couple of steps, he stopped and glanced back at Mainstein.
"Mannstein, have you discovered something?"
"We might have found something, or we might not have found anything. I'll let you know once I see the staining results."
Fritz didn't ask any more questions. He went into the slicing room and closed the door. Mainstein stood in the middle of the animal room, looking around. M7 rolled over in its cage, from lying on its back to lying on its side, its hind legs kicking in its sleep, as if it were walking. Several other experimental monkeys were in cages nearby, some sleeping, some idly gripping the bars, showing no interest in Mainstein's presence. The innermost cage was now empty; it belonged to M8. M8 had been moved to the post-operative observation room yesterday, its spinal cord sliced into thousands of thin sections and stored in a freezer at -80 degrees Celsius, waiting to be analyzed layer by layer by Mainstein.
M21, the "surprise," had already completed its experimental journey months earlier. On the day it was euthanized, Fritz spent a long time alone in the animal room. He squatted in front of M21's cage, whispered something in German, and then wiped the cage clean, making it look brand new. When Mainstein entered the animal room, Fritz was pushing M21's cage to the corner. Mainstein asked him what he was doing, and Fritz said, "Making room; M21 won't be coming back." Mainstein didn't stop him, but he never let a new monkey into that cage. The cage remained empty, with a label on the door that read "M21."
An empty cage is more unpleasant than a full one.
Mainstein walked to the cage where M21 had been and stood there for a while. The handwriting on the label was Fritz's, neat and clear, each letter written carefully. "M21. Non-targeted intervention group. Unexpected recovery." Unexpected recovery. Mainstein looked at these four words and suddenly felt they were very inaccurate. This was not an accident. M21's recovery, like M7's recovery, and like Chen Jianguo's recovery, were all different manifestations of the same biological law. It's just that it took them longer to understand what M21 was telling them.
M21 was an outlier from the start. In the non-targeted intervention group, it was the only monkey to show functional recovery. The other five showed nothing: flat electromyography, zero behavioral scores, and no signs of repair, bridging, or regeneration after dissection. Only M21 began to show voluntary contraction of the hind limb muscles at the sixteenth week post-surgery, regained spontaneous urination at the twenty-second week, and stood with assistance at the twenty-eighth week. Mannstein didn't know how to explain this phenomenon at the time. 62% of the monkeys in the targeted intervention group recovered function, while only M21 in the non-targeted intervention group did. Statistically, this is called "occasional," biologically, it's "worthy of further investigation."
After leaving the slicing room, Mainstein went directly to Yang Ping's office.
Yang Ping is resting.
"what happened?"
"Professor, I need you to confirm something for me."
"what's up?"
Mainstein walked in, opened the folder, and inside was a stack of printed photomicrographs. He laid the photos out one by one on Yang Ping's desk, arranging them in a row. Each photo was an immunofluorescence staining image of a spinal cord slice; red dicortin labeled newly formed neural progenitor cells, green NeuroD labeled differentiating neurons, and blue DAPI labeled all cell nuclei.
“This is a spinal cord slice from M21. I had Fritz re-stain it, using double-labeled corticosteroids and NeuroD.” Mainstein pointed to the first photograph. “Look at the area around the injury.”
Yang Ping leaned closer and examined the photographs carefully. Around the center of the injury, along the pathways of blood vessel distribution, there were densely packed red and green fluorescent signals. The red and green signals overlapped to form yellow, which was the area where dicortin and NeuroD were co-expressed, and where neural progenitor cells were migrating and differentiating.
“This is almost identical to the slice of M8,” Yang Ping said.
"Almost identical, M21 did not undergo precise gene editing; we only made extensive microenvironmental adjustments, without even using a targeting vector. But progenitor cells were still activated in its spinal cord. This shows that activating progenitor cells does not require precise gene editing, but only a microenvironment that allows them to grow, and the substance that initiates this behavior is within that microenvironment."
Yang Ping picked up a photograph and examined it under the light for a while. Clusters of red cells were distributed along the course of small blood vessels, extending from the periphery of the injury towards the center, like an army on the march. Their shapes were very irregular, with large nuclei and prominent nucleoli, completely different from the surrounding mature neurons.
“This discovery is more important than the discovery of M8.” Yang Ping put down the photograph.
"why?"
"Because M8 received a complete intervention—targeted vector, gene editing, and precise regulation—you can say in your paper, 'Our method activated the progenitor cell.' But M21 didn't receive any of that. It only received a non-specific microenvironmental adjustment. If it can also activate the progenitor cell, it means that the conditions for activating the progenitor cell may be more complex than we thought, and there are other pathways besides precise gene editing."
Mainstein looked at Yang Ping, his eyes lighting up. "Professor, do you mean our method is just one of many activation methods? Perhaps there are other simpler and safer ways to achieve the same effect?"
“Possibly! But not necessarily.” Yang Ping placed the photo back on the table, crossing his arms over his chest. “While the non-targeted intervention of M21 is not precise, it still alters the microenvironment. It doesn’t do nothing; it does something similar to M8, just in a different way. The real question is, what exactly is the key signal that activates the progenitor cells? Is it a specific molecule? Or a physical condition? Or both? What we see now is the result, not the mechanism. The mechanism still requires extensive experimental exploration. However, based on my experience with K therapy, it is very likely a specific molecule that the human body often uses as a key to unlock certain locked behaviors.”
Mainstein was silent for a moment. He knew Yang Ping was right; there was a world of difference between seeing the results and understanding the mechanism. What did M21 tell them? Progenitor cells could be activated by non-specific microenvironmental modulation. But M21 couldn't answer the questions of "how to activate" and "why it can activate."
"So what's our next step?" Mainstein asked.
"We need to create a complete molecular atlas from the M21 slices. We need to do everything we can: single-cell sequencing, spatial transcriptomics, proteomics. We need to figure out where these activated progenitor cells come from—are they local adult stem cells, precursor cells that migrated from the blood, or transdifferentiated from surrounding glial cells? Then we need to go back to the mouse model and conduct systematic mechanistic studies. We need to use chemical inhibitors to block possible signaling pathways one by one and find that indispensable key molecule. Also, we need to verify this in Chen Jianguo. If we can detect dicortin-positive cells in his cerebrospinal fluid, it means that this mechanism is conserved in humans."
Mannstein jotted down notes quickly in his notebook. His handwriting was so messy that only he could understand it.
"Professor, how do you verify this on Chen Jianguo? I don't want to do a spinal cord puncture; the risks are too great."
"A small amount of cerebrospinal fluid is obtained through lumbar puncture to detect the concentrations of dicortin and NeuroD. If the protocellular cells are activated, they will release some factors into the cerebrospinal fluid. We don't need to see the cells themselves, just their fingerprints. Don't forget, the cerebrospinal fluid circulation itself is a transport pathway."
When to do it?
"The sooner the better, preferably tomorrow."
Mannstein closed his notebook, stood up, walked to the door, then stopped. "Professor, do you know what I'm thinking right now?"
"what?"
“I was thinking about M21, it’s called ‘Surprise.’ When we named it that, we just thought its recovery was an unexpected, inexplicable phenomenon. Looking back now, it wasn’t unexpected. It was an answer that arrived ahead of time, it just took us months to understand it.” Yang Ping didn’t answer, he lowered his head, his gaze falling on the photomicrographs. Those red, migrating, repairing cells were working quietly in M21’s spinal cord, like a group of tireless craftsmen. They were unknown to anyone while M21 was alive, only discovered by Mainstein in the corner of thousands of slides after M21 died. That’s how science is, ninety-nine percent of discoveries come from deliberate design. One percent comes from accidents. And that one percent of accidents is often more valuable than the ninety-nine percent of designs.
Mannstein went to the rehabilitation room.
Chen Jianguo was doing standing exercises. He held onto the parallel bars, his legs trembling slightly, and beads of sweat glistened on his forehead under the light. Sister Li stood behind him, holding a towel, ready to wipe away his sweat at any moment.
“Professor Mainstein.” Chen Jianguo immediately stopped.
"Actually, you can call me Professor Mann! Stand still, I'm here to tell you something."
Chen Jianguo steadied himself on the parallel bars, took a deep breath, and before he could speak, Mainstein said:
"I need to take some cerebrospinal fluid from you."
Chen Jianguo was taken aback. He had spent two years in a rehabilitation hospital and knew what cerebrospinal fluid was—the fluid that surrounds the brain and spinal cord, extracted via lumbar puncture. He also knew what a lumbar puncture felt like: a long needle piercing through the back, passing through the intervertebral space, and entering the subarachnoid space. It didn't hurt, but it was very uncomfortable.
"When?"
"tomorrow."
"it is good!"
The next day, cerebrospinal fluid was successfully extracted. Clara divided the sample into three parts: one for routine biochemical testing, one for ELISA to detect the concentrations of dicortin and NeuroD, and one for centrifugation to collect the precipitate for cell smear preparation. The cell smears were ready the following morning. Clara fixed, stained, and mounted the smears and placed them under a microscope.
She saw a few scattered cells. Their shape was peculiar: large nuclei, sparse cytoplasm, and tiny protrusions along the edge of their cell membranes. Clara stared at these cells for a long time, then stood up and went to find Mainstein.
"Professor Mainstein, come and take a look."
Mainstein walked over and approached the microscope. He adjusted the focus, located the cells in the field of view, and remained silent for a long time. "Was the double corticosteroid staining done?"
"It was tested, and the result was positive."
What about NeuroD?
"I also took the test, and it came back positive."
Mainstein straightened up, leaned against the lab table, and looked at Clara. "This means that Chen Jianguo's cerebrospinal fluid contains cells that are double-positive for both corticosteroids and NeuroD. These cells did not enter from outside the blood-brain barrier; they detached from the spinal cord and entered the cerebrospinal fluid. This indicates that the activation and differentiation of progenitor cells are underway in his spinal cord."
Should we tell Professor Yang?
"Yes, but not now. Let's repeat it first to confirm it's not contamination. Take another sample of cerebrospinal fluid, centrifuge it again, and stain it again. If we see the same cells, we'll tell Professor Yang."
Clara repeated the test, and the result was the same. Double corticosteroid positive, NeuroD positive, and the cell morphology was exactly the same as before. Mainstein took both sets of results to Yang Ping's office.
After reading the two reports, Yang Ping placed them on the table, leaned back in his chair, and stared at the ceiling.
"Professor, you guessed it?"
"We were speculating when the M21 slice results came out. Now we have found evidence that it also applies to humans."
Mainstein sat down and looked at Yang Ping. What could he say? His theory was correct, his predictions were correct, and every act of persistence he made was right. But Mainstein knew that Yang Ping wouldn't say something like, "I was right." Yang Ping would only ever talk about what to do next, not what he had done right in the past.
"Professor, we now have three layers of evidence. The first layer is the M7 primate acute injury model. The second layer is the M8 primate chronic injury model. The third layer is the unexpected recovery from non-targeted intervention in M21. Now we have Chen Jianguo's cerebrospinal fluid, and the fourth layer is clinical evidence in humans. Four layers of evidence, from rodents to primates to humans, from acute to chronic to non-targeted. This theory is no longer a theory; it is a fact."
Yang Ping took out the folded piece of white paper from his pocket; it was the initial manuscript of his three-dimensional guided gene theory. The paper was almost torn at the creases. He spread it out on the table, looking at the dense handwriting, the arrows and question marks, and the sentences that had been crossed out and rewritten.
"Professor, you still keep this?"
"Keep it. Sometimes you can look back and see what you were thinking back then, reminding yourself not to go astray. Some things have never changed."
What has remained unchanged?
"Actually, the core of the whole theory lies in two key points—the desired cells appearing in the desired location! One can be explained by the three-dimensional spatial guidance gene theory, and the other can be explained by the stem cell theory. The two theories should actually be unified at the underlying level. I am now thinking about whether we need to find this more fundamental unified theory."
Mannstein looked at Yang Ping, deeply shocked, his heart racing uncontrollably. He knew just how significant this unified theory truly was.
"You're saying that stem cells and three-dimensional guided genes are actually a unified theory, just like quantum mechanics and classical mechanics can be unified?"
"Yes, the human body repeats this process from the moment of conception, ensuring that the desired cells appear in the desired locations; none of these can be omitted. I suspect that regulating cell differentiation and positioning is actually a set of mechanisms."
"This is such a wonderful surprise, Professor!"
"That's true to some extent!"