ABSTRACT
Although the water-soluble metabolite profile of human mesenchymal stem cells is known, the lipid profile still needs further investigation. In this study, methanol-chloroform was used to extract pid-soluble metabolites and perchloric acid was used to extract water-soluble metabolites. Furthermore, a dual phase extraction method using methanol-chloroform and water was used to obtain both water and lipid fractions simultaneously. All metabolite extractions were analyzed on a 9.4T high-resolution nuclear magnetic resonance spectrometer. Metabolite resonance peaks were assigned in the acquired spectra according to the chemical shift, and the extraction efficiency of ferent methods was compared. Results showed that in the spectra of water-soluble extracts, major metabolites comprised low molecular weight metabolites, including lactate, acetic acid, fatty acids, threonine, glutamic acid, creatine, choline and its derivatives, while in the spectra of lipid-soluble extracts, most metabolites were assigned to fatty acids. Among the different extraction procedures, perchloric acid was more efficient in extracting water-soluble metabolites and methanol-chloroform was efficient in extracting organic components compared with the dual phase extraction method. Nuclear magnetic resonance spectroscopy showed that as low as 0.7 mg organic yield was enough to obtain clear resonance peaks, while about 6.0 mg water-soluble yield was needed to obtain relatively favorable spectral lines. These results show that the efficiency of extracting water and lipid fractions is higher using perchloric acid and methanol-chloroform compared with dual phase extraction and that nuclear magnetic resonance spectroscopy is highly sensitive for analyzing lipid-soluble extracts.
ABSTRACT
OBJECTIVE: To observe the characteristic of the fMRI brain map in patients undergoing needling at Zusanli (ST36) by reinforcing method for exploring the essence of Meridian-Collaterals and the mechanisms of acupuncture in treating diseases. METHODS: Twenty-six healthy volunteers were randomly assigned to two groups by double blinded method, 16 in the acupoint group and 10 in the non-acupoint group. Using GE Signa 1. 5 T superconducting MRI system, the fMRI was performed with Gradient echo-EPI sequence. Post-processing of fMRI data was performed using the Functool software (GE-ADW4.0) to generate positive correlation coefficient brain functional activating images and the data was analyzed statistically using SPSS 13.0 software. RESULTS: Brain functional area was elicited in 13 out of the 15 patients in the acupoint group and 10 in the non-acupoint group. Among them, the temporal elicited area in the acupoint group showed specificity (Fisher's Exact test, P = 0.022) and only the difference in contralateral hemisphere activation rate was of statistical significance (McNemer test, P = 0.020). CONCLUSION: Acupoints has its own specific brain activated areas. The therapeutic effect of acupoint might be mediated through brain to treat diseases and regulate functional disorder of organs. There exists special transmission channel of meridian.
