Regenerative medicine is a technology that uses normal cells and tissues to treat organs and human tissues that have lost their functions due to diseases and injuries. Regenerative medicine can be roughly divided into several methods, such as culturing epidermis, cartilage and flaky myocardial cells, cell recombination, injecting cells into human body, using cell drugs and so on. At present, there are four kinds of regenerative medical products approved by Japanese Medical Device Law (Pharmaceutical Machinery Law) and included in the scope of insurance treatment. Among them, there are three kinds of products using tissue engineering technologies such as cell regeneration and recombination, namely "Jace" products for treating burns, which are patient epidermal cell slices cultured in vitro introduced by J-TEC (Japanese tissue engineering) company; The patient's chondrocytes were cultured, encapsulated in polymer gel and transplanted into the joint "Jack" product. TERUMO's "heart slice" product transplants muscle cells from patients with severe heart failure to the surface of the heart.
In terms of cell drugs, JCR Pharmaceticals has launched the product of "TEMCELL HS Note". Taking bone marrow mesenchymal stem cells as effective components can effectively control the immune response after leukemia hematopoietic stem cell transplantation.
Whether it is a venture enterprise or a large pharmaceutical enterprise, the research and development technology of regenerative medical products in Japan has developed rapidly.
"Immune checkpoint inhibitor" helps immune T cells to identify cancer cells that have escaped the human immune response, and uses T cells to attack cancer cells to achieve therapeutic purposes.
The human immune system can identify and eliminate foreign bodies. As a part of the immune system, an immune cell called cytotoxic T cell is mainly responsible for recognizing and attacking foreign bodies. Of course, in order to avoid excessive immune attack on autologous cells, the human immune system keeps a pathway to inhibit immune response, which is the "immune checkpoint".
Immune checkpoint inhibitor is a new type of anticancer drug, which can block immune checkpoints and stimulate cytotoxic T cells to attack cancer cells. Cancer cells are very cunning and will use the mechanism of immune checkpoints to cleverly avoid the attack of immune T cells.
Representative immune checkpoint inhibitors are ONO PHARMACEUTI-CAL-Cal-Cal's "Opdivo" by Xiaoye and MSD's "Keytruda" by Merck kgaa in the United States. Drugs such as Opdivo and Keytruda bind to "PD 1" immune checkpoint molecules on the surface of cytotoxic T cells, which hinders the combination of PDL 1 and PD 1 in some cancer cells, thus removing the restriction of immune response.
Opdivo and other drugs have played an amazing role in the treatment of some cancers, and companies have also joined the development camp of immune checkpoint inhibitors, and the competition is becoming increasingly fierce. Similar to Opdivo, drugs combined with PDL/kloc-0 and/or other immune checkpoint molecules have been successfully developed.
After cancer cells are infected with oncolytic virus, the virus will multiply rapidly and eventually dissolve the cancer cells. After the cancer cells are dissolved and destroyed, the oncolytic virus will spread outside the cell and continue to infect the next cancer cell. This will also activate the body's own immune function. If used together with popular cancer treatment drugs such as Opdivo, the treatment will get twice the result with half the effort.
Oncolytic viruses can change and reconfigure the genes of many viruses, such as adenovirus that causes colds and herpes virus that causes herpes simplex infection. These characteristics can prevent cells other than cancer cells from being infected by viruses, and even if infected, it is difficult to reproduce.
In 20 15, Amgen's IMLYGIC was officially approved. Since then, some large pharmaceutical companies have taken actions to obtain the drug technology and sales rights developed by venture enterprises.
Japan is also developing related technologies. Oncolys BioPharma Company has made great achievements in the research of oncolytic virus, developed Telomelysin, and recruited patients with esophageal cancer in Japan on 20 17 to start clinical trials.
Chimeric antigen receptor T cell immunotherapy (CART therapy) is a cell therapy that transforms immune cells into aggressive cells and destroys cancer cells.
The mainstream of CART therapy uses cancer patients' own T cells. Specifically, firstly, an immune cell named "T cell" (blue cell in the figure) was isolated from the blood of cancer patients, and the "chimeric antigen receptor" (orange part in Figure 3-4) gene was embedded in the T cell. T cells containing components only respond to cancer cells and have the function of attacking immune cells of cancer cells. Increase the number of "super aggressive" T cells and then re-enter the patient's body. At this time, the super-invasive T cells returned to the patient's body give full play to the invasive function of cancer cells, at the same time enhance the cell activity and continue to proliferate, ensuring long-term high invasive ability.
2065438+At the end of August, 2007, the chimeric antigen receptor T cell therapy (CART therapy) developed by Novartis Company of Switzerland was approved by the United States for the first time.
After CART treatment, most patients' conditions were controlled. Novartis takes a life-threatening leukemia as the experimental object, takes the marker of cancer cells as the target, and adopts CART therapy to treat it. After 3 months of medication, it was found that almost all the cancer cells in 83% patients disappeared.
In addition, in the treatment of malignant lymphoma, the venture capital company Kite Pharma has submitted an application for CART therapy to the United States. In Japan, Novartis Pharmaceutical Co., Ltd., a Japanese legal person, jointly developed CART therapy for severe leukemia and malignant lymphoma with Baori Medical Biotechnology Company and No.13 Company. Accompanied by the over-attack effect is the side effect of cart therapy. After clinical application, how to quickly find and deal with side effects has also become an urgent task. In addition, the cart therapy at this stage is "tailor-made", and the production and distribution costs are high. In the future, relevant parties should not only consider how to reduce costs, but also study the payment of medical expenses from the social level.
A gentle spray on the site where cancer may occur will only make cancer cells glow in a few minutes. This is the "cancer fluorescent spray". In the near future, cancer fluorescent spray may appear in the medical field as a sharp weapon to assist endoscopy and surgery.
In order to apply this spray to the "intraoperative rapid pathological diagnosis" technology of breast cancer, the drug was approved on 20 18, and the performance evaluation of cancer fluorescent spray has been carried out comprehensively. Endoscopic examination and surgical safety test of esophageal cancer have also begun.
The scientific name of this spray is "fluorescent probe", which was jointly developed by Professor Yasuhiro Urano from the Department of Pharmacy and Medicine of the Graduate School of Tokyo University and Professor Kobayashi Jiulong from the National Institutes of Health (NIH). The reagent will emit fluorescence after reacting with some proteolytic enzymes, and its main component is small organic molecules.
Fluorescent probe is a reagent that binds amino acids and rhodamine fluorescent molecules. Under normal circumstances, it is colorless and non-fluorescent. After the reagent meets the proteolytic enzyme on the surface of cancer cells, the fluorescent molecules decomposed by water immediately dissociate amino acids, enter the inside of cancer cells and emit fluorescence. As long as less than 1 mg is sprayed on the suspected cancer place, the cancer place will light up within a few minutes.
The important field of clinical research of this reagent is breast cancer. In order to avoid residual lesions, breast cancer surgery needs to make slices (sections) to detect whether cancer cells have been completely eliminated, which is called "intraoperative rapid pathological diagnosis". Fluorescent probe technology can make diagnosis quickly and is an important means to reduce the burden on surgeons and pathologists.
So far, fluorescent probe technology has achieved more than 90% verification accuracy, which can clearly identify breast cancer. Taking Fukuoka General Hospital of Jisheng Association (Fukuoka City) as the center, many institutions are conducting clinical research on breast cancer and collecting data for a whole year. According to the requirements, when applying to the PMDA Drug and Medical Device Administration for drug clinical trials, relevant materials must be submitted. If it is soon, the fluorescent probe will apply for drug access on 20 18.
In breast cancer surgery, in order to protect the integrity of breast morphology, many patients choose partial resection, but partial resection also increases the risk of cancer residue. In order to check whether there is any residue, it is necessary to carry out "intraoperative rapid pathological diagnosis", but many medical institutions are faced with the problems of insufficient pathologists and large business volume, which is difficult to fully implement.
Both Koryo Chemical and Hamamatsu Photon have joined the research camp. Professor Urano of the University of Tokyo authorized Pentagonal Chemicals to manufacture fluorescent probes, while Hamamatsu Optoelectronics began to develop a device for quantitative measurement of fluorescence intensity.
"In-vivo hospital" is a technology for human body to make self-diagnosis and treatment on necessary occasions and at necessary time.
Nano-molecules, known as "intelligent nano-machines", wander around the human body and diagnose and treat diseases such as cancer on the spot. The Nano-medical Innovation Center, with the main goal of "in vivo hospital", was selected as COINS Project, the innovation output project stronghold of the Ministry of Education, Culture, Sports, Science and Technology of Japan, and its director is Kataoka.
In order to realize intelligent nano-machine technology, Kataoka and others developed a drug delivery system for cancer. Using hydrophilic and hydrophobic polymers as tissues, the drugs were directly wrapped in nanocapsules (polymer micelles) to treat the affected areas.
The research and development of anticancer drugs coated with polymer micelles has condensed everyone's efforts. It is necessary to design the diameter of polymer micelle to be 30 nm and the virus size to be 100 nm. Only in this way can it be ensured that it will not enter the vascular space of normal tissue, but can enter the large space space unique to cancer tissue. Only in this way can we ensure the targeted effect of drugs on cancer.
The PH value (hydrogen ion index) of cancer tissue is lower than that of normal tissue. After the reaction, the polymer micelle was broken and the anticancer drugs in it were released. Polymer micelles enter cancer tissues like Trojans and launch fierce attacks. Many enterprises are developing polymer micelle technology to wrap anticancer drugs, and clinical trials are under way.
Encapsulation of anticancer drugs with polymer micelles is the first step to realize intelligent nano-machine technology. In the second step, Kataoka and others are working hard to develop drugs with both diagnostic and therapeutic effects. One of the achievements is "nano-mechanical contrast agent", which is beneficial to visually check the malignant and refractory parts of cancer through MRI. Under the action of gastric acid, the nanoparticles coated with manganese contrast agent only react with the unique environment of cancer and release the contrast agent.
Kataoka believes that the ultimate goal of nano-machine technology is to collect all the biological information in the patient's body and feed it back to the chip built in the body, so as to complete the disease diagnosis. It can be said that this idea is similar to the structure of an asteroid detector. Maybe one day in the future, the world described in the sci-fi movie Fantasy Journey half a century ago will really come true.
"Virtual colonoscopy" uses multi-slice spiral CT (computed tomography) to take pictures of the large intestine, and makes three-dimensional images of the large intestine through computer processing to help doctors find polyps and cancer lesions. Also known as "CT colonoscopy".
Virtual colonoscopy uses multi-slice CT with more than 16 rows to accurately photograph the peristalsis of large intestine in a short time, and this technology has been applied in clinic. After many slice cross-sectional images taken by multi-row CT are combined into three-dimensional images, the observation effect is almost the same as that of endoscope, so this technology is also called "virtual endoscope".
Through clinical observation and research, virtual colonoscopy is as sensitive and specific as endoscopy in finding lesions, and many in-depth medical institutions have begun to introduce virtual colonoscopy. The large intestine has many folds and curved shapes. After using virtual colonoscopy, even the lesions hidden in folds can be accurately found.
In the process of CT examination, a small amount of radiation is inevitable. According to the introduction of National Cancer Research Center of Japan, after simulating the whole virtual colonoscopy process, the radiation dose at the two positions is 2-3mSv, which is about1/5 (10-12 msv) of the radiation dose detected by enema X-ray.
At present, in the examination of colorectal cancer, it is necessary to carry out fecal occult blood test on patients first, and then carry out colonoscopy after confirming positive. Considering many factors such as taking laxatives, complicated pretreatment process, shame and so on, women tend to stay away from endoscopy. Moreover, in the actual testing process, only about 30% people really need to test. Moreover, when the endoscope is inserted from the anus and then pulled out, it is difficult to find the hidden lesions hidden in the folds of the large intestine.
Intestinal bacterial therapy is a therapeutic method to inject intestinal flora into large intestine, regulate intestinal environment, and treat and prevent diseases. Studies have shown that the disorder of normal flora in intestinal flora is the main cause of diarrhea, constipation and obesity. Recently, some research results have proved that intestinal flora will not only lead to difficult diseases such as ulcerative colitis and allergic enteritis, but also induce many diseases such as nervous system diseases and coronary artery diseases.
The injection of intestinal bacteria can be divided into the following categories: intestinal transplantation of feces, capsule transplantation without bacteria in the intestine, and drug administration to treat intestinal flora diseases.
Many medical institutions in Japan have conducted clinical trials and research on fecal transplantation therapy, and the research object is elderly inpatients who are prone to infection with difficult intestinal infectious diseases and ulcerative colitis. Among them, the research team of Shuntian University mainly studied the fecal transplantation and the combined treatment of antibacterial drugs for patients with ulcerative colitis. After taking antibacterial drugs, the number of intestinal flora is greatly reduced, while after transplanting feces, the intestinal flora is greatly improved.
During the treatment, after taking antibacterial drugs, about 200 grams of normal saline was added to the patient's feces collected that day to make a solution of about 400 ml, and the solution was injected into the appendix. The diagnosis was made by colonoscopy within 6 hours after transplantation.
So far, in clinical research, about 80% of patients who have completed the treatment have significantly improved their symptoms. After analyzing the intestinal flora, the researchers found that compared with the ineffective flora, the proportion of bacteria-"bacteroides", which is the main component of effective flora, increased significantly, indicating that the intestinal flora of patients gradually stabilized.
In the future, the research team of Shuntian University plans to treat Crohn's disease with fecal transplantation and antibiotics. The intestinal flora of Crohn's disease patients is very disordered.
"Non-invasive continuous blood glucose detection" is a detection method (non-invasive) for directly measuring blood glucose changes without blood sampling. In this method, sensors are installed in the subcutaneous tissue of the patient's abdomen and wrist, and the fluctuation of blood sugar value is simulated by measuring the glucose current conversion of tissue fluid.
20 17, 1 year 10 month, the "free Libre" product for patients to measure blood sugar at any time came out, and it was included in the Japanese insurance category in September. This product is sold by Abbott Japan. Using "FreeStyle Libre" can directly measure the blood sugar data of 14 days in real time, without blood collection. The feature of "FreeStyle Libre" product is that patients manage their own machines without doctors. After the sensor is installed in the human body, the patient can immediately know the blood sugar data at that time and the fluctuation of blood sugar value as long as he touches the sensor with a reader. This product is beneficial to prevent hypoglycemia, reasonably control diet, and control the rise of blood sugar. It can also remind users to improvise during exercise, and may even change the traditional treatment of diabetes.
Before the listing of "FreeStyle Libre", Abbott released the "FreeStyle Libre Pro" product on 20 16 12. This product is for doctors only, and the longest measurement time is 14 days. Some experts said: "When the monitoring time is two weeks, the dosage and types of drugs can be adjusted every week, and the most suitable prescription can be given to patients after analyzing the blood sugar results." This product has many advantages, not only can continuously record the changes of patients' blood sugar, but also can help to find patients' hypoglycemia at night.
Both products are designed with minimum current fluctuation, and there is no need to prick the finger to correct the value. In the past, most products need to puncture the fingertips to collect blood, which is an invasive detection method.
"Angiography" is mainly used for the diagnosis of cardiovascular diseases such as angina pectoris, which can measure the number, distribution and shape of atherosclerosis and whether there is tear in the intima of blood vessels.
In recent years, the development of "intravascular endoscopy" is particularly rapid, and there is also the technology of "intravascular ultrasound (IVUS)" which uses ultrasound to observe vascular sectional images in real time. Neither of these two techniques requires X-ray examination, so patients do not need to worry about the influence of radiation, and it is also convenient for doctors to observe. This technology began to be used in clinic in 1990s, and the technological innovation has been continuously improved.
A major technological innovation of vascular endoscope comes from JIMRO, a subsidiary of Otsuka Holdings. 2065438+May 2007, the company released a brand-new vascular endoscope "Angiography IJS 2.2". The new product adopts 3 MOS camera and LED light source, and the output image is high-definition and perfect.
Another technological innovation of vascular endoscope is "double perfusion". Naturally, the coronary artery is needless to say, and even the aorta with a lot of blood flow is clearly visible. The new technology is helpful for doctors to observe the subtle injury of aorta, such as the precursor of aortic dissection, which is difficult to diagnose so far.
In IVUS, the catheter is inserted directly through the vascular lesion, and the tip of the catheter is equipped with an ultrasonic transceiver, which can slowly take images of the lesion. The frequency of ultrasonic wave has increased from 40MHz to 60MHz in the past, and new products with greatly improved resolution and shortened detection time have also come out.
High-resolution technology is helpful to clearly see the separation of atherosclerotic arteries on the inner wall of blood vessels, and it is also easy to evaluate the regeneration of implanted stents to improve arterial stenosis. The examination time is short, the coronary artery insertion time is reduced, and the risk of ischemia is greatly reduced.
In the past, when diagnosing ischemic cardiovascular diseases such as angina pectoris and myocardial infarction, it was necessary to fill the vascular cavity with contrast agent and perform coronary angiography with X-ray irradiation. Patients are not only irradiated, but also unable to detect the shape and development of atherosclerotic arteries.
Protein (nuclease) has the function of scissors and is used to cut off various biological genes (DNA). In the process of gene repair, the genetic factors of cells are modified by changing DNA sequences, or similar DNA sequences are replaced, and DNA sequences from other organisms are implanted from the broken parts. This is gene editing technology. With gene editing technology, human beings can freely change the genes of species, develop new foods and drugs, and their applications in the biological field are also expanding.
Up to now, gene editing technology has gone through three generations: the first generation is "zinc finger nuclease (ZFN)" and the second generation is "transcription activator-like effector nuclease (TALEN)". The third generation is "clustered regular staggered short palindrome repeat sequence (CRISPR/Cas 9)". Among them, CRISPR/Cas 9 technology can complete gene editing in a short time, which is cheap and quickly popular all over the world. Using CRISPR/Cas 9 technology, human beings can change the genes of plants, fish, nematodes, mice, pigs, monkeys and humans, and the universal applicability of this technology has also accelerated its popularization.
Many countries use CRISPR/Cas 9 technology to cultivate transgenic animals and carry out experiments such as recombinant cells. This technology is not only used in real life, but also cultivated many excellent varieties by collecting high and small cells produced by substances and carrying out gene therapy. And it has blossomed in agriculture, forestry, fishing, chemistry, medicine and other fields.
For example, CRISPR/Cas 9 technology can be used to change the genes that inhibit muscle growth and cultivate pigs, snappers and so on. They are fat and strong with plenty of edible parts. In addition, the study of removing the abnormal gene of congenital amaurosis (LCA) is also under way.
In the past, transgenic technology generally used to irradiate multiple individuals with radiation to change their genetic characteristics, select individuals (mutants) who unexpectedly mutated after irradiation and met the requirements, extract similar DNA sequences for homologous recombination, and embed the genetic fragments that need to be introduced.
Taking the cultivation of transgenic knockout mice as an example, the cost of homologous recombination needs 3 million to 5 million yen, and the time is 1-2 years. With the emergence of CRISPR/Cas 9 technology, the cost is only a few thousand yen, and the time is shortened to about one month.
The "New Generation Mini-sequencing Technology" is a small device for reading genetic factors and sequencing genome bases at high speed.
In 20 15, the British Oxford Nanohole Technology Company launched a product called MinION all over the world. MinION is only the size of a palm and is connected to a personal computer. The company provides the host computer free of charge, and users only need to buy 1 thousand dollars 1 disposable sensor. Because of its small size, it can be used outdoors. In order to reuse water in space, the National Aeronautics and Space Administration (NASA) introduced MinION to measure water pollution.
Oxford Nanopore Technology will release smaller and cheaper products after 20 17. As the number of sensors reading genome (DNA) and ribonucleic acid (RNA) is reduced, the cost of the disposable part is reduced by 1/3- 1/5. Looking around the world, not only Oxford Nano-pore Technology Company has a new generation of small sequencing technology, but also Japanese Quantum Biological Systems Company is developing related technologies. We look forward to a more active market in the future.
Genetic factors carry the protein information of various functions of organisms, and there are countless genetic factors in the genome. The causes of intractable diseases and the development of new drugs are inseparable from the analysis of genetic factors and genomes.
The total amount of information in the genome is different for different biological species. There are about 3 billion base pairs in the human genome, so the detection of such a huge genome has to rely on the support of high-speed reading technology and "new generation of small sequencing technology". Reading the base information from many genome fragments and searching for the read fragment information on the Internet can get the genome sequencing of protozoa.
The technology of high-speed and large-scale data analysis is rapidly popularized, but the introduction cost ranges from tens of millions of yen to hundreds of millions of yen, which is too expensive. However, in order to increase the information of gene fragments and carry out optical detection of fluorescent markers, large equipment is essential. MinION uses a special protein sensor to measure the current through unit DNA and RNA, and then completes the gene analysis. Because the CCD camera and laser technology used to read genes are simplified, the size of the equipment is also smaller.
The technology of "frozen electron microscope" is to put the measured object such as biomolecules in an ultra-low temperature environment of about MINUS 200 degrees Celsius, take images by using electron beams, and analyze them by computer, and finally obtain the tiny three-dimensional structure of the measured object. The English name of Cryo in cryomicroscope is Cryo-electronic Microscopy, which means ultra-low temperature. Since the end of 20 13, it has attracted attention from all walks of life.
The resolution of cryoelectron microscope is1Amy (0.1nm), which is close to the size of a single atom and can accurately analyze the three-dimensional structure of biomolecules such as protein. It is of great benefit to the development of drugs and other products after unraveling the biomolecular structure of biomolecules and the "culprit" of infection. If we can solve the molecular structure of plant photosynthesis, we can even artificially complete photosynthesis and synthesize organic matter from sunlight.
The specific application steps of frozen electron microscope technology are as follows: First, the particle structure of biomolecules such as protein is interrupted, and frozen samples embedded with extreme particles are made. The size of each protein molecule is about 10 nanometer, and the frozen sample can accommodate multiple particles. Place the sample in a frozen environment for observation. In one night, the equipment can automatically take hundreds of high-resolution electron microscope images. There are hundreds of particles photographed in a single image, so the total number of protein particles photographed in one night will exceed 654.38+ million. Tens of thousands of complete and good data can be screened and analyzed by computer, and finally more detailed three-dimensional structural information can be obtained.
If the sample is of good quality, the atoms that make up the molecule can even be observed by an electron microscope. After observing 1 week, an image with a resolution of 5 Amy can be obtained, and an atomic model can be obtained in about one month.
1 cryoelectron microscope needs an investment of1-200 million yen, and many research institutions, universities and pharmaceutical and chemical enterprises have introduced it one after another. The well-known company in cryoelectron microscope industry is JEOL in Japan.
Before the appearance of cryogenic microscope, scientists mainly analyzed the structure of biomolecules by crystal X-ray diffraction analysis. After the crystal is irradiated with X-rays, the X-rays will be diffracted with different internal densities of the crystal. The three-dimensional structure of the crystal is analyzed by using physical principles. The more regular the crystallization, the larger the volume and the more detailed the stereo structure information. However, it is difficult to obtain high-quality crystals of biomolecules, so there are still many mysteries about the three-dimensional structure of protein. The industry also expects to analyze the structure of more amorphous protein by cryoelectron microscopy.
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