1September, 1988 to 199 1 July, 1998, studied for a master's degree in human anatomy at Sun Yat-sen University of Medical Sciences. Under the guidance of Professor Tang Tingyong, he completed his master's thesis "Aging changes of arcuate nucleus of hypothalamus in rats". It is found that the neurons of hypothalamic arcuate nucleus, glial cells, synapses and β -endorphin neurons in aged rats change with age, which is the physiological function of hypothalamic-pituitary axis in the elderly. Hippocampus is closely related to learning and memory, and the learning and memory ability of elderly individuals and patients with Alzheimer's disease is declining. We studied the development of hippocampus and excitatory amino acid neurons in detail, which provided abundant morphological data for brain aging.
2. Peripheral nerve injury and its repair
Supported by two projects of the Provincial Health Department, we studied the effects of injecting basic fibroblast growth factor into target muscle on the regeneration and functional recovery of sciatic nerve injury, the effects of human embryonic spinal cord extract on the survival and growth of motor neurons in the anterior horn of spinal cord of embryonic rats, and the effects of embryo motor neuron transplantation on denervated muscles. It provides a new route of administration for the clinical application of basic fibroblast growth factor.
3. Experimental study on molecular biological diagnosis methods of Alzheimer's Harmo disease and Alzheimer's Harmo disease.
⑴ Study on the genetic and molecular biological diagnosis of Alzheimer's disease.
At present, the early diagnosis of AD is still a difficult problem in neuropsychiatry. So far, we have not found a biological diagnostic index that fully meets the requirements, so it is very meaningful to find an effective biological index for prenatal diagnosis and early diagnosis. It can not only diagnose AD disease early, guide clinical treatment plan and strive for early treatment, but also make it possible for different research institutions to carry out comparative research because of the application of the same biochemical indicators, which lays a foundation for further clarifying the pathogenesis of AD disease. At present, with the support of the State and Guangdong Natural Science Foundation, we are carrying out genetic and molecular biological diagnosis research of Alzheimer's disease, and have achieved good results.
① A fluorescent quantitative RT-PCR method was successfully established to detect APP and HO- 1 genes.
A fluorescence quantitative RT-PCR method for detecting APP and HO- 1 genes was successfully established. It was found that APP and HO- 1 genes were highly expressed in the cells of patients with Alzheimer's disease, which was related to the severity of AD. In the next step, a large number of samples will be detected and compared with other detection methods to explore whether detecting APP and HO- 1 genes are expected to become biological indicators for detecting AD.
② The correlation between ②ApoE gene polymorphism and D 10S 1225 locus and Alzheimer's disease.
One or more susceptibility loci on the long arm of chromosome 10 are associated with late-onset Alzheimer's disease (Lod). Through family linkage analysis, the maximum multi-point dominant logarithm score near D 10S 1225 was obtained, suggesting the possible etiological significance of this locus to the dominant logarithm. We successfully established a reverse dot blot hybridization method for ApoE genotyping (China patent application number: 2004 100525307), and genotyped D 10S 1225 by polyacrylamide gel electrophoresis and silver staining. It was found that the frequency distribution of ApoE allele ε4 in the case group was significantly higher than that in the control group. Seven alleles were detected at D 10S 1225 * *, and the sizes were 178bp,18/bp, 184bp and184bp respectively. It is suggested that ApoE allele ε4 is the susceptible factor of load, and the results show that there may be no susceptible gene linked to load imbalance near D 10S 1225.
③ Neuroprotective effect of human telomerase reverse transcriptase gene transfection on Alzheimer's disease and its mechanism.
The full-length cDNA of hTERT was obtained from pCI-neo-hTERT by PCR, and was directionally cloned into pADTrack-CMV shuttle plasmid. PADEasy- 1 and recombinant shuttle plasmid (pADTrack- hTERT) were transformed into BJ5 183 by step transformation for homologous recombination. HEK293 cells were transfected with hTERT recombinant adenovirus skeleton plasmid (pAD- hTERT), and the expression of green fluorescent protein (GFP) was observed by fluorescence microscope. The DNA extracted from recombinant adenovirus (AD-hTERT) was harvested for PCR identification and amplification. PCR, restriction endonuclease digestion and sequencing confirmed that hTERT cDNA was correctly inserted into shuttle plasmid and recombined with pADEasy- 1 PCR identification and reporter gene analysis showed that hTERT recombinant adenovirus was packaged correctly and had good infection activity. The successful construction of hTERT recombinant adenovirus laid a foundation for the study of hTERT's neuroprotective effect.
⑵ Experimental study on the treatment of Alzheimer's disease with traditional Chinese medicine
In view of the important role of β -amyloid protein (Aβ) in the pathogenesis of Alzheimer's disease, the research on the treatment of Alzheimer's disease with Aβ as the target was carried out, and the mechanism of the effective component TA990 1 extracted from natural plants affecting the aggregation and fiber formation of Aβ peptide was studied in series, and the curative effect and mechanism of TA990 1 in preventing and treating Alzheimer's disease were expounded. And obtained the invention patent of China (patent number ZL065438). We also found that EGb76 1 can greatly enhance the function of TA990 1 and has a synergistic effect. Based on the drug screening model of fluorescence spectrophotometry, the quantitative structure-activity relationship and molecular mechanism of the compatibility of two drugs (TA9902) were studied by spectrometry. It is found that TA9902 can better inhibit the aggregation and fiber formation of Ab. Cell culture found that TA9902 also has neurotrophic effect, which can promote the survival and process growth of neurons in cerebral cortex and alleviate the neurotoxicity caused by Aβ. This research was supported by the National Natural Science Foundation of China and several provincial funds, and obtained 1 national patents. More than 65,438+00 papers have been published in core journals. In 2003, he won the second prize for outstanding academic papers in natural science in Guangdong Province and the third prize for scientific and technological progress in Guangdong Province in 2004. It will have an important impact on the research and development of a class II new drug for treating Alzheimer's Harmo's disease and a traditional Chinese medicine for preventing and treating AD with independent intellectual property rights, and bring huge economic benefits and benefits.
⑶ Experimental study on the treatment of Alzheimer's disease with vaccine.
With the support of the State, Guangdong Natural Science Foundation and key scientific and technological projects and cross-cutting projects in Guangzhou, and under the leadership of Professor Yao Zhibin, we carried out experimental research on vaccine treatment of Alzheimer's disease. Using Aβ42 peptide as vaccine for pathological intervention immunotherapy of AD is a breakthrough in foreign AD treatment research in recent years and a revolution in the treatment of non-communicable diseases. We have successfully constructed a subunit vaccine of Aβ42 (China invention patent application No.03 1 14042.4). Aβ subunit vaccine can effectively induce normal SD rats, BALB/c mice, AD transgenic mice (Tg2576) and adult rhesus monkeys to produce specific anti-Aβ42 antibodies with high titer, without adverse reactions, showing good immunogenicity and safety. This research was supported by the National Natural Science Foundation and several provincial and Guangzhou key funds, and more than 30 papers were published in domestic and foreign journals. Our goal is to develop cheap and effective new drugs to treat AD within 3 to 5 years.
4. Targeted diagnosis and treatment of nanoparticles
He undertook the research on the sub-topic "The distribution of nanoparticles in bone matrix-induced heterotopic bone formation and human osteosarcoma metastasis model" of the national high-tech research and development plan (863) project "Targeted diagnosis and treatment of nanoparticles" (No.:2001A218031).
In recent years, he has presided over important academic exchange conferences and continuing education program classes at home and abroad, made special reports and read papers for more than 20 times.