Analysis on the origin and development of elements of reinforcing and reducing needling techniques / 针刺研究

Reinforcing and reducing needling manipulations are important factors affecting clinical therapeutic effect. In the present paper, the relevant elements of the reinforcing and reducing techniques of acupuncture needle including the left- and right-ward twirling, gender, needling at the left and right, front and back parts of the body, needling along or against the running course of the meridian, and their origin and development recorded in ancient Chinese medical books were collected and sorted out, followed by analysis on the understandings of Chinese ancient medical practitioners about them. Results show that the right- or left-ward twirling of needles, gender, and needling at the right or left part, the front or the back part of the body of patients are not the core components of the reinforcing and reducing techniques. Of the three stimulus parameters of needling, named amplitude, frequency and duration which are frequently researched at present, only the duration of single twirling (frequency) was highly noted in GAO Wu's book Zhenjiu Juying (A Collection of Gems in Acupuncture and Moxibustion). It is worthy of being studied in the further. Regarding the stimulation intensity of acupuncture involving the identification of reinforcing or reducing manipulations, the factors influencing the patients' feelings of needling intensity of acupuncture should be studied at the same time.

Crystallization separation of antitumor constituent 20(R)-25-OCH3-PPD / 中草药

Objective: To separate antitumor constituent AD-1 [20(R)-25-OCH3-protopanoxadiol (PPD)] from non-racemic enatiomeric mixture. Methods: A crystallization method has been developed to separate AD-1 from 25-OCH3-PPD non-racemic enatiomeric mixture, the sorts and amount of solvent and crystallization temperature were optimized to obtain the best separative conditions, and the relationship between the S/R percentage of 25-OCH3-PPD non-racemic enantiomeric mixture and dielectric constant of the solvent was discussed. Results: Under the optimized conditions (acetone 26 mL, 4°C), the yield and purity of AD-1 crystals are 38.76% and 97.83% from 25-OCH3-PPD non-racemic enatiomeric mixture (R: 59.34%; S: 38.35%), respectively. Conclusion: The ε values of solvents are about 20.7, and an unsaturated solution and low temperature are beneficial to the separation of AD-1 from 25-OCH3-PPD non-racemic enatiomeric mixture.

Construction of eukaryotic recombinant expression plasmids with glyceraldehydes-3-phosphate dehydrogenase and cysteine protease inhibitor gene of periodic Brugia malayi and its expression in HeLa cells / 中国地方病学杂志

Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.

Cloning, sequencing of CPI gene from periodic Brugia malayi and prediction of B cell epitopes in its amino acid sequence / 中国地方病学杂志

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.