The development of biochemistry and molecular biology in China

Abstract

Originally, biochemistry or physiological chemistry was a preclinical course in medical schools. At the beginning of the last century, medical schools were established all over China. Among them, one of the best known was Peking Union Medical College (PUMC) founded in 1917 with the endowment of Rockefeller Foundation. Its Department of Physiological Chemistry was established in 1919, headed by Hsien Wu in 1924 and he renamed the Department of Biochemistry in 1925. From 1920 to 1941, Wu and his colleagues in PUMC carried out pioneering work on blood analysis, protein nutrition, protein denaturation, immunochemistry, and so forth. His system of blood analysis marked the beginning of clinical chemistry. His book Principles of Nutrition published in 1929 was the first of its kind written in Chinese. His theory of denaturation based on a series of thorough studies was published in the Chinese Journal of Physiology in 1931 (1) and was republished in Advances in Protein Chemistry in 1995 (2) because the theory he proposed 64 years ago is surprisingly consistent with our present knowledge of protein structure. Through biochemical studies in his laboratory, he trained many visiting scholars and graduate students, and they afterwards became the leaders of biochemistry in other medical schools. Unfortunately, this golden period ended after Japan had invaded China in 1937 and occupied PUMC in 1942 after the Pacific War. During the anti-Japanese War, most universities and medical colleges moved to Southwest China but biochemical studies did not stop, even under very harsh conditions. For instance, the Department of Biochemistry in Central University headed by Ji Zheng (Libin T. Cheng) continued to do nutrition research in Chengdu. In 1938 and 1945, respectively, Bang-Zhe Ren (P. C. Jen) coming back from H. B. Lewis' lab in Ann Arbor and Ying-Lai Wang from Keilin's lab in Cambridge joined him, thus, making his lab a biochemical research center in the vast unoccupied area. In teaching, he had trained both undergraduate and graduate students using Bodansky's Physiological Chemistry and A Laboratory Manual of Biochemistry written by himself, which was also widely used in other medical schools. Before retirement, he was Professor and Head of the Department of Biochemistry at Nanjing University where he trained numerous outstanding biochemists. Now he is enjoying his retirement at the age of 110. Another active biochemical research center during the war was the Faculty of Science in Southwest Union University where biologists studying zoology, botany, plant physiology, and plant pathology gathered to continue their studies and publish their articles in the Biochemical Bulletin, an English language journal edited by the famous plant physiologist Pei-Song Tang and printed with a mimeograph. Prof. Hsien Wu (1893–1959) After the Second World War ended in 1945, exiled universities and medical colleges moved back to resume their normal teaching and research. After my graduation from Zhejiang University in 1948, I came back to my native city Changsha and worked in Hsiang-Ya or Hunan-Yale Medical College (HYMC) for a couple of years as an assistant in the Department of Biochemistry headed by P. C. Jen. In HYMC, biochemistry was taught in English using Benjamin Harrow's Textbook of Biochemistry. In the laboratory, I practiced blood analysis following Hsien Wu's protocol to prepare a protein-free blood filtrate and determine the blood sugar content using a DuBosq visual colorimeter. At that time, I had the opportunity to read a new book, Dynamic Aspects of Biochemistry written by Earnest Baldwin and published in 1947. This book was a gift to P. C. Jen from H. B. Lewis. As a newcomer to biochemistry, I benefited greatly from reading this concise and up-to-date book. I believe other biochemists reading this book also found it very inspiring. After the war, PUMC reopened in 1948. Its Department of Biochemistry was headed by William Adolf until he left for the United States in 1950. He had worked in China for over 20 years and educated many students of biochemistry. Ying-Lai Wang was one of his graduate students in Yenching University. Before he left, I spent about 6 months in his lab to purify a plant protein from Phaseolus vulgaris. After Adolf's leaving, Zhi-Quan Liang (C. C. Liang) who obtained his PhD from the State University of Pennsylvania came back in 1950. He co-chaired in 1951 and chaired in 1958 the Department of Biochemistry in PUMC. Robert K. S. Lim, a famous physiologist who directed the Department of Physiology at PUMC from 1924 to 1937 planned to set up a Medical Institute in Shanghai belonging to the Central Academy of Sciences and De-Pei Feng (T. P. Feng) who used to work with Lim in PUMC was appointed in 1944 as the Director of the Medical Institute in Preparation. In 1948, Ying-Lai Wang also moved from Central University to the Medical Institute in Preparation. On the basis of the Medical Institute in Preparation, the Institute of Physiology and Biochemistry belonging to the Chinese Academy of Sciences was established in 1950 and divided into the Institute of Physiology and the Institute of Biochemistry in 1958. The development of biochemistry before 1949 was fully reviewed in A History of the Early Development of Biochemistry in China (1917–1949) written by Libin T. Cheng in Chinese and published by Nanjing University Press in 1989 (3). It is a valuable book highly recommended to those who read Chinese. The development of biochemistry and molecular biology from 1949 to 2008 can be subdivided into two periods, each spanning about 30 years before and after China was opened up. Biochemical research from 1949 to 1988 was reviewed in Current Biochemical Research in China edited by C. L. Tsou and published by Academic Press in 1989 (4). After Nixon's visit in 1972, Jordan Tang at the Oklahoma Medical Research Foundation and Yuan-Chuan Lee at Johns Hopkins University were among the early American scientists visiting China. After coming back, they wrote an article, Biochemistry in the People's Republic of China published in Trends in Biochemical Sciences February 1980 (5). This article reviewed some aspects of biochemical research and teaching in China before China's opening up. After 1949, the peaceful and stable environment favored the development of science and there was a surge of scientists returning from abroad. To promote the development of science, the Chinese Academy of Sciences was established in November 1949, right after the founding of New China in October 1949. In 1956, the Chinese Academy of Medical Sciences was also established. Many students were sent to the Soviet Union and obtained Candidate Degrees after their graduate studies. There was no education of graduate students in China after 1949 until Premier Zhou started “Marching toward Science” in 1956. In 1957, I was fortunate to pass the entrance examination and study protein chemistry in the Institute of Physiology and Biochemistry as a graduate student of Tian-Qin Cao (Tian-Chin Tsao). In the 50s and 60s of the last century, biochemical studies were mainly carried out in research institutes. In universities and medical colleges, most professors were busy with teaching and did not have enough time to do research. Zhi-Quan Liang and his colleagues at the Institute of Basic Medical Sciences, belonging to the Chinese Academy of Medical Sciences, were mainly concerned with protein research and later nucleic acids as well. In protein research, they developed plasma substitutes from bovine serum albumin. The Institute of Biochemistry directed by Ying-Lai Wang was mainly concerned with proteins, enzymes and nucleic acids. In enzymology, Chen-Lu Tsou (C.-L. Tsou) and Tsing-Ying Wang in collaboration with Ying-Lai Wang succeeded in the purification of membrane bound succinic dehydrogenase and found that its FAD prosthetic group was covalently linked to the enzyme protein (6). In 1962, Chen-Lu Tsou proposed a graphical method for the determination of the number and type of essential groups in protein molecules according to the relations between the fraction of activity remaining and the modification of a certain type of functional group (7). In comparison with Koshland's kinetic method, Tsou's method should have a much wider application. In muscle protein research, Tian-Chin Tsao et al. studied a series of tropomyosins and paramyosins from a variety of sources. In 1963, Tian-Chin Tsao, Tsu-Hsun Kung, et al. did pioneering electron microscopic studies on tropomyosin and paramyosin paracrystals (8). Tsao's group also studied plant viruses infecting tobacco, wheat and other crops in collaboration with many labs of plant pathology. Ying-Lai Wang (1907–2001) Chen-Lu Tsou (1923–2006) Tian-Chin Tsao (1920–1995) Studies on nucleic acids were retarded in the Soviet Union and China by Lysenko's anti-Mendelian doctrines, though rapid progress in molecular and cell genetics proved that nucleic acid was the structural basis of heredity. In 1956, at a Tribune on Genetics convened in Qingdao by the Chinese Academy of Sciences and the Ministry of Higher Education, Mendel–Morgan's genetics were rehabilitated. De-Bao Wang (T. P. Wang), just coming back from the United States to the Institute of Biochemistry was an invited speaker at the meeting. Since then, he had led a group studying nucleic acids, especially tRNA. The achievements in nucleic acid research in De-Bao Wang's lab and Zhi-Quan Liang's lab before the Cultural Revolution were reviewed by T. H. Cheng and R. H. Doi in Recent Nucleic Acid Research in China and published in Progress in Nucleic Acid Research and Molecular Biology in 1968 (9). Zhi-Quan Liang (1914–2006) De-Bao Wang (1918–2002) This tour de force project started in 1958 and succeeded in 1965, before the “Cultural Revolution.” It was a collaboration of three research teams. The Institute of Organic Chemistry and the Department of Chemistry of Peking University were responsible for the synthesis of the A-chain. The Institute of Biochemistry was responsible for the synthesis of B-chain, the combination of A- and B-chain of natural insulin to obtain re-synthesized crystalline insulin and finally the combination of synthetic A- and B-chain to obtain totally synthetic crystalline insulin. Conventional peptide synthesis in solution was used to make the A- and B-chains. The re-synthesis of natural insulin in crystalline form from its two chains not only paved the way for the total synthesis of insulin but also demonstrated that the three-dimensional structure of insulin was determined by its primary structure. The totally synthetic insulin was fully characterized by physicochemical analyses and biological assays. The structural analysis of porcine insulin crystal by the Peking Insulin Structure Research Group marked the beginning of protein crystallography in China. This project, started in 1967, was directly stimulated by the total synthesis of bovine insulin. In 1971 and 1973, the three dimensional structure of crystalline insulin was solved at 2.5 and 1.8 Å resolution respectively (11). Through this work, long-term collaboration between this group and Dorothy Hodgkin's group at Oxford University was established. In 1972, Dorothy Hodgkin brought her electron density map of insulin to Beijing and compared it with the electron density map there. Afterwards, she wrote a short note, Chinese Work on Insulin in Nature May 1975, anticipating closer exchange of insulin research between the East and the West (12). Another reaction to the total synthesis of insulin was the total synthesis of yeast alanine tRNA composed of 76 nucleotides. This project was started in 1968 and completed in 1981. It was carried out by researchers from the Shanghai Institute of Biochemistry, the Shanghai Institute of Cell Biology, the Shanghai Institute of Organic Chemistry, the Institute of Biophysics, the Department of Biology of Beijing University, and the Shanghai Second Reagent Factory. Six fragments of oligonucleotides were synthesized by a combination of chemical and enzymatic method and joined by RNA ligase to form two half molecules. Finally, the two synthetic half molecules were joined by RNA ligase to obtain the totally synthetic tRNA molecule containing all modified bases and exhibiting full biological activity. This work is a nation wide collaboration involving many institutes and hospitals. The anti-malaria artemisinin was discovered in 1971 by You-You Tu et al. at the Institute of Chinese Medical Sciences in collaboration with other institutes in Shangdong and Yunan. It was extracted in crystalline form from a Chinese herb Artemisia annua (Qinghao) and proved to be highly effective in the treatment of malaria, especially pernicious malaria. Its structure was solved by Wei-Shan Zhou et al. at the Institute of Organic Chemistry in 1976 and published in Chinese in Acta Chimica Sinica May 1979 (14). Later, its crystal structure was solved in the Institute of Biophysics. Artemisinin is a sesquiterpene compound containing 15 carbon, 22 hydrogen, and five oxygen atoms but contains no nitrogen, thus differing from all other nitrogen containing anti-malaria drugs. On the basis of this structure, derivatives with improved action were synthesized by Ying Li et al. at the Institute of Materia Medica in Shanghai, among them artemether being one of the best. This achievement has had a huge impact in the development of effective drugs from Chinese Traditional Medicine. The World Health Organization has recommended an artemisinin-based combination therapy (ACT) to all countries where the malaria parasite has developed resistance to chloroquine. Here, I have to stress that the aforementioned formidable projects before China's opening up were carried out from scratch through the collaboration of different laboratories where only primitive facilities were available and many reagents were self-prepared. The development of biochemistry and molecular biology was greatly affected by the chaotic “Cultural Revolution” which was started in 1966 and eventually ended in 1976. During this decade, there was tremendous progress elsewhere in molecular biology, especially molecular cloning and genetic engineering. After 1979 when China adopted the open door policy, research activities gradually returned to normal. To catch up with developments abroad, many people were sent to Western countries to study as visiting scholars or graduate students. In January 1979 when China and the United States established diplomatic relations, American laboratories began to accept Chinese visiting scholars. In 1981, a CUSBEA (China-United States Biochemistry Examination and Application) Program was initiated by Ray Wu, son of Hsien Wu, at Cornell University to help young people in China study as PhD students in first rate American universities. Up to 1989, 422 graduate students were sent to the United States through this program. As more and more people studied abroad and came back, research activities were not only centered in academies and institutes but also actively carried out in the universities. The most urgent effort was to catch up with the rest of the world. Here, I would just mention some of the most important achievements. The cloning and expression of HBV surface antigen gene subtype adr was achieved by Zai-ping Li et al. at the Institute of Biochemistry. The HBV genomic DNA was cloned and sequenced, and then the surface antigen gene was cloned to vaccinia virus of low toxicity. Instead of using the recombinant vaccinia virus directly as a live vaccine as reported by B. Moss et al. at NIH, the recombinant virus was grown in chicken embryo and the expressed surface antigen was purified to form 22 nm particles, which could be used as a safe hepatitis B vaccine. The cloning of the hearing impairment gene GJB3 was accomplished by Jia-Hui Xia et al. at the National Medical Genetics Lab in collaboration with medical doctors and published in Nature Genetics December 1998 (16). The GJB3 gene encoding the human gap junction protein was mapped to chromosome 1p33-p35. It was shown that a mis-sense mutation and a nonsense mutation were associated with high frequency hearing loss in two families, thus realizing the cloning of the first disease associated gene in China. Since then, other genes associated with various diseases, such as dentinogenesis imperfecta, brachydactyly, and atrial fibrillation, have been cloned. The aforementioned two projects are noteworthy as they represent the so called “zero-breaking” (meaning from none to available in a certain field) achievements. With increasing financial capabilities, China decided to join the expensive Human Genome Project in September 1999 and began the sequencing of a region in the short arm of chromosome 3 containing about 30 million base pairs or 1% of the whole genome. This task was accomplished in time through the joint effort of three genome centers: the Human Genome Center at the Institute of Genetics led by Huan-Ming Yang, the Chinese National Human Genome Center at Beijing led by Bo–Qin and the Chinese National Human Genome Center at Shanghai led by Zhu Chen. This project was initiated by Dong-Cai Liang and Pei-Song Tang because of its importance in photosynthesis and carried out by Wen-Rui Chang et al. at the Institute of Biophysics and Ting-Yun Kuang et al. at the Institute of Botany. It did not succeed until 2003 and was published in Nature March 2004 (17) Although thousands of protein crystal structures were solved, yet the structural analysis of membrane proteins was still a challenge. The major light-harvesting complex of photosynthesis II is the principal solar energy collector of green plants and may also function in photo-protection under high light conditions. The structural biology laboratory of the Institute of Biophysics was involved in the solving of the crystal structure of insulin and that of light-harvesting complexes in blue-green algae. The accumulation in protein crystallography and the experience of growing protein crystals in this laboratory laid the foundation for the successful solving of the first membrane protein in China. Later, the crystal structure of another membrane protein—the mitochondrial respiratory complex II was solved and published in Cell July 2005 (18) through the collaboration of Tsinghua University and the Institute of Biophysics led by Zi-He Rao. The modification of DNA by sulfur in Streptomyces lividans 1326 was discovered by Zi-Xin Deng et al. at Shanghai Jiaotong University and was first published in Molecular Microbiology September 2005 (19). The entire dnd gene cluster involved in this modification was localized on an 8 kb DNA fragment. In addition, the same DNA modification associated with the Dnd (DNA degradation) phenotype was also observed in widespread bacterial species. Further studies revealed that this site-specific modification was an isosteric replacement of oxygen by sulfur resulting in the phosphorothiolation of DNA, the first known physiological modification of the DNA backbone. This important discovery in basic research should have important implications in cell biology. In the last 10 years, with the rapid development of economy in China, the investment in science and education has increased exponentially. Now, the research facilities are equivalent to or even better than those abroad. Recently, the Shanghai Synchrotron Radiation Facility was built in Pudong New District by the Shanghai Institute of Applied Physics in collaboration with the Institute of High Energy Physics in Beijing. In August 2008, the facility ran successfully beams at 3.5 GeV in the storage ring with super conducting RF cavities. Furthermore, most components were made by the above two institutes. With improved working conditions and much higher payment to scientists, more and more overseas Chinese scientists are attracted to develop their careers in China. Last year, more than five thousand scholars including biologists came back to work in Shanghai. In all, about 300,000 overseas Chinese came back to China. In this period, so many achievements in biochemistry and molecular biology have been made in China, that it is impossible to describe them in a limited space. Fortunately, important articles of well-established researchers in China were published in international journals and should be familiar to their colleagues abroad. In addition, the 21st Congress of IUBMB will be held in Shanghai in August 2009. On that occasion, there will be an all-round display of the recent progress in this field of biology in China and in other countries. This also gives me the excuse that, to describe the current development of biochemistry and molecular biology in China is not only beyond my ability but also unnecessary. The history of a biochemical society in China can be traced back to 1926 when the Chinese Physiological Society was founded by Robert KS Lim and Hsien Wu. The first generation of Chinese biochemists joined this society and published their papers in the Chinese Journal of Physiology. Before 1949, Chengdu Biochemical Society and Shanghai Biochemical Society were established in 1938 and 1948, respectively. The Chinese Biochemical Society was first established in Shanghai in December 1949. There were only 36 members in the beginning and later the number did not increase very much. During the ‘Cultural Revolution’, the activities of all societies were stopped. After China's opening up, the Chinese Biochemical Society was reestablished in May 1979 during the First National Conference of Biochemistry held in Hangzhou. After two months, a Chinese delegation of biochemists joined the 11th International Congress of Biochemistry held in July 1979 in Toronto. In 1993, the Chinese Biochemical Society adopted its present name, the Chinese Society of Biochemistry and Molecular Biology. China joined the IUB in 1958. In 1961, the IUB Council accepted the application of Taiwan to join the Union. This was unacceptable to China, as Taiwan is an integral part of China. In protest, China withdrew from the Union. In July 1979, a Chinese delegation of six members headed by Ying-Lai Wang joined the 11th International Congress of Biochemistry in Toronto. During the meeting, Ying-Lai Wang and Cheng-Lu Tsou discussed with IUB General Secretary W. J. Whelan and Treasurer E. C. Slater about the restoration of China's membership in IUB. As a result of protracted negotiations, the Chinese Biochemical Society rejoined the Union. With the cooperation of J. C. Su and T. B. Lo from Taipei, the Biochemical Society located in Taipei, China also joined the Union (20). I was fortunate to be one of the six members in the Chinese delegation and witnessed this event. This win–win solution promoted the scientific exchange between Chinese biochemists across the channel and their relations with the world communities. The same kind of solution was adopted by other scientific unions and later by ICSU itself. Now, after 30 years, the 21st International Congress of Biochemistry and Molecular Biology will be held in Shanghai. As its host, the Chinese Society of Biochemistry and Molecular Biology will make every effort to ensure the success of this global meeting.