Analysis to Next Generation Sequencing Technology

With the development of science, traditional Sanger sequencing has failed to meet new requirements of low cost, high throughput and fast in speed.

Recent years, with the discovery and promotion of second-generation sequencing technology, the gene sequencing speed has increased greatly while achieving a substantial decline in costs, making large-scale application of genome sequencing possible. Now, the cost of personal whole genome sequencing is about 5,000$, and is expected to decreased to less than $ 1,000 in the next few years.

The rapid development of sequencing technology will promote the massive accumulation of DNA sequencing data, accompanied by the accumulation of the corresponding individual diseases, signs and other data at the same time. When we accumulate enough data, how to understand these data will be critical. On the micro level, generations of molecular biologists’ studying the effects of apparent biological traits genes exert on utilizing technologies like gene knockout have made breakthroughs in many crucial aspects. On the macro level, statistics and other data analysis techniques are introduced to study the relationship between gene sequences and biological phenotype. The accumulation of basic scientific research gradually brings breakthroughs in clinical applications.

There are now two types of clinical applications mainly, one aims at disease screening of ordinary people. It infers people’s future risks of getting cancer by measuring the known genes associated with a disease loci. The other aims at the diagnosis cancer and other deadly diseases. It finds in a series of drugs or plans the most effective one for certain patients by testing the loci of certain genes.

Data from BBC research shows that total global gene sequencing market increased from $ 7.941million in 2007 to $ 4.5 billion in 2013, and will reach $ 11.7 billion in the year of 2018 with the CAGR up to 21.2%.

Currently, the market of de novo sequence platform is mainly taken by several major manufacturers, including the Illumina, Ion Torrent / Life Technologies (was the acquisition of Thermo Fisher in 2014), 454 Life Sciences / Roche, etc.

Under such a circumstance, the next generation sequencing technology (second-generation sequencing) appears. As an emerging industry, the next-generation sequencing technology can be applied in clinical testing like antibody discovery, health industry, industrial and agricultural use of gene-oriented study as well as scientific research and development.

Discussion on Cell Therapy From The Point of Standardization, Scale, and Industrialization

What is cell therapy?
Cell therapy refers to the transplantation or input of normal or bioengineered human cells into a patient’s body and newly-imported cells can replace damaged cells or involve a stronger immune killing function, so as to achieve the purpose of treating diseases. Cell therapy has shown higher application value in the treatment of cancer, hematological diseases, cardiovascular diseases, diabetes, Alzheimer’s disease etc. In general, cell therapy includes tumor cell immunotherapy and stem cell therapy. There are two cell sources for cell therapy, one from the patient itself and the other from the allogeneic tissue.

The Defects of Cell Therapy
The cell is the most basic unit that contributes to a living organism, however, it does not mean that everyone shares the same cells. On the contrary, there is a huge difference in each individual which can be compared to human-to-human differences, that is, two identical people never exist. The huge difference between cells and cell preparations is the biggest drawback of cell therapy. In this post, we will discuss several issues that need attention in the current stage of cell therapy.

Difficulties in the Standardization of Cell Therapy
Cancer cell immunotherapy cannot be standardized from the stage of raw material acquisition. The cell treatment materiasl for each paitient are their own blood leukocytes. The condition and physical condition of each patient are different, and the collected white blood cell growth quantity and kill activity are not uniform and cannot be standardized. As it is impossible to standardize raw materials, preparation processes, and product specifications, it cannot be standardized, industrialized, and scaled up. Each tumor cell immunotherapy laboratory meets the GMP level with the hardware environment, and it can be more like a cell preparation workshop. Researchers ranged in number from a few to a dozen and could not really meet the standards of division of labor of industrialized pharmaceutical companies. Taking stem cell therapy that using umbilical cord mesenchymal stem cells as an example, which raw material is an umbilical cord, and one umbilical cord-produced cell can be utilized by many paitients. The standardization path is more advanced than the immunotherapy of tumor cells, and the raw materials can be standardized to some extent.

Difficulties in The Scale of Cell Therapy Industry
At present, the production mode of the cell therapy industry mainly depends on technicians. In the 10,000-grade clean laboratory, the cells are operated in class 100 clean bench, cultured in a carbon dioxide incubator, centrifuged in a centrifuge, observed through an inverted microscope, and the drug reagents are stored in a medicine refrigerator. All of these devices are operated by independent biological laboratories of the individual and being linked together through the operations of scientists. This type of production model is small in scale and similar to workshop-type production. Although there are some large scales, the essence is a collection of many small workshops. Due to the small scale, the instruments used are laboratory instruments and many of the reagents used are scientific reagents, which will lead to the issue of low efficiency but high cost.

Autologous or Allogeneic
There are two kinds of cell sources for cell therapy, one from the patients and the other from the allogeneic tissue. Autologous cell therapy cannot be standardized from the raw material acquisition stage, and it are only applied to the patient itself, the essence is essentially medical technology. The prevalence of autologous cell therapy as a medical technology is mainly due to the scale of the predicament. Allogeneic therapy, the cells derived from allogeneic. Taking tumor cell immunotherapy as an example, the cell source may be from cord blood, and the larger-scale cell source may be a filter plate for leukocyte filtration at the blood bank. Taking umbilical cord mesenchymal stem as an example, the cell source is the umbilical cord, and one umbilical cord-producing cell can be used by more than one person. If scale can be cultivated, although the quality standards cannot be quantified well, the scaled products themselves have a certain degree of standardized properties.

The cell industry, as an industry, is not the path to the advancement of cell-based therapeutics. If the advanced technology cannot be mass-produced on a large scale, it can only stay in the laboratory and become the object of research for scientists, never have achance to become a drug into the majority of patients. For allogeneic cell therapy that using allogeneic cells as raw materials, the standardized properties of the scaled products can be realized if large-scale cultures are prepared, then scale and standardization can promote each other. The current progress in standardization of cells is not easy, but the progress in scale should be relatively easy to achieve.

Natural cytokine supernatants with more standardized and standardized properties
Cytokines are a class of small molecule proteins with broad biological activity synthesized and secreted by immune cells (such as monocytes, macrophages, T cells, B cells, NK cells, etc.) and certain non-immune cells (endothelial cells, epidermal cells, fibroblasts, etc.) Immune responses are regulated by binding to the respective receptors to regulate cell growth, differentiation and effects. Cytokines (CK) are low-molecular-weight soluble proteins that are produced by various types of cells induced by immunogens, mitogens, or other stimulants. They have the ability to regulate innate immunity [1] and adaptive immunity [2], hematopoiesis, cell growth, and damage tissue repair and other functions.

Cytokines can be divided into interleukins, interferons, tumor necrosis factor superfamily, colony stimulating factors, chemokines, growth factors etc. Cytokines form a very complex cytokine regulatory network in the body and participate in many important physiological functions of the human body. Where stem cells and immune cells cannot reach the body, cytokines can easily reach target tissue sites because of their small size.

In recent years, recombinant gene cytokines have made remarkable achievements in clinical applications as a novel biological response modifier. A large part of the effects of stem cell therapy and immune therapy arises from the action of cytokines secreted in the body. The stem cells and immune cells in the body are introduced back into the body to secrete a variety of natural structural cytokines. Although the amount of these cytokines is relatively small, they are synergistic and act directly on the cytokine network in the body because of their high natural structure activity, lack of antigenicity but diversity. Because of the standardization, standardization, industrialization, and scale of natural compound cytokines, it is more cost-effective than cell therapy, allowing more patients in need to enjoy cell-like therapeutic effects.

Although natural complex cytokines can largely replace cell therapy, but there are still conditions that require the presence of cells to exert a therapeutic effect. We hope that cell therapy can break the current situation, become high efficiency and low cost with large scale, more standardization, and then be applied to more disease treatments.

A Healthy Dose of Medicine for the Soul

A large segment of the human population takes things way too seriously for their own good. The strange anomaly is that most people laugh at the wrong thing and fail to laugh at the right thing. This serious incongruity has robbed people of a healthy attitude towards life in general.

Those who take life too seriously are in danger of missing the great joys of living in a crazy world like ours. I am not sure about the scientific research but I would guess that for every sad moment it takes one hundred laughs to balance the books. Some people are about ninety-nine laughs short of a real sane moment.

I like the old English proverb that says, “Laugh and the whole world laughs with you, cry and you cry alone.”

From my perspective, if you cannot laugh with someone you will not be able to cry with him or her and have it mean anything.

According to some medical advice, it takes more facial muscles and energy to frown than it does to smile. Of course, the only exercise some people have is frowning and who am I to take that away from them.

I am determined, no matter what, to exercise my right to smile and laugh and enjoy the world around me. I must confess that I get this attitude quite honestly.

My paternal grandfather was a Past Master in the area of practical jokes. No amount of time was too much to spend preparing for one of his famous practical jokes. His favorite holiday was April 1 and began preparing for this holiday right after Christmas.

The fact that his practical jokes at times got him into trouble did not seem to affect him at all.

Once while in the hospital for an extended period he had somebody smuggle in to him a can of snuff. For some reason he liked chewing snuff. It is the most disgusting habit I know of on earth.

He no sooner received his smuggled goods then he began chewing it. If you know anything about chewing snuff, you know it is accompanied by a lot of spitting. As usual, his timing was impeccable. Just as the head nurse passed his door and looked in, he leaned over and spit in to the garbage can he had next to his bed. The nurse, not knowing about the chewing snuff, thought he was spitting blood and immediately went into emergency mode. Immediately my grandfather was rushed into the operating room and the surgeon and medical team were assembled.

My grandfather was very sick at the time. Some did not think he would get out of the hospital.

Just as they got him situated in the operating room he pulled from under his sheet his can of chewing snuff and smiled at them. The only person in the room that thought this was in any way amusing was my grandfather. The doctors were so angry with him that they refused to see him for three days and confiscated his can of chewing snuff.

My aunt and uncle lived right next to my grandfather. My aunt was hyper clean when it came to her house. Dirt in any form was not welcome under her roof. She had a broom that was always within reach because she never knew when a piece of dirt would try to invade her domicile.

That year my grandfather found something new. I am not sure where he found it but he probably spent a lot of time looking for something like this. It was a rubber facsimile of a very nasty looking piece of vomit. To him it was a prized possession.

Most of his practical jokes were executed on April 1. Whenever we saw grandfather coming on this particular day we usually ran for cover.

He went over to visit my aunt and was sitting on the couch in the living room. They chatted for a little while and then my grandfather began to cough a little bit. He said to my aunt, “I haven’t been feeling good lately. I really don’t know what it is.” Then he started to cough a little more seriously, to which, my aunt got up and went to the kitchen to get him a glass of water thinking that might help him.

When she got back, she was shocked to see on her new coffee table a very horrible sight. My grandfather was bent over the coffee table hacking and coughing as though he was in the process of dying. On the coffee table was a very nasty looking piece of vomit.

My aunt went into hysterics. She whirled around and within a moment had grabbed her broom and started towards my grandfather. My grandfather was laughing but not for long.

Suddenly he realized that the flailing broom in my aunt’s hand was aimed at him. She chased him out of the house, down the driveway and for at least three blocks yelling obscenities at him that I dare not repeat in public society.