To control the quality of Maca, the quality standard was established in this study. According to the methods recorded in the Appendix of Chinese Pharmacopoeia (2010 Edition), the water, extract, total ash, acid insoluble substance, and heavy metals inspections in Lepidium meyenii were carried out. N-benzyl-9Z, 12Z-octadecadienamide in L. meyenii was identified by TLC, and it was determined by HPLC. The results showed that the N-benzyl-9Z, 12Z-octadecadienamide identification of TLC was a strong mark and specificity. In content determination experiment, the linearity of N-benzyl-9Z, 12Z-octadecadienamide was in the range of 0.01-2 microg (r = 0.9998), and the average recovery (n=9) was 99.27% (RSD 2.0%). The methods were simple, accurate, with good reproducibility. It is suitable for quality control L. meyenii.
Chromatography, High Pressure Liquid ; methods ; Chromatography, Thin Layer ; methods ; Lepidium ; chemistry ; Plant Extracts ; chemistry

0.05),higher rate of maternal mortality and perinatal mortality(P

Objective To observe the effect of early rehabilitation treatment at different time point on the motor function and activity of daily living (ADL) and emotion in acute stroke patients. Methods 120 patients with acute stroke were randomly assigned to three reha-bilitation groups and one control group according to the disease course (3 days, 5 days, 8 days when the disease is steady). Each group con-sisted of 30 eases. The patients in each group were treated with the regular medication therapy, rehabilitation groups were treated with com-prehensive rehabilitation treatment including Bobath technique therapy. Neurological deficit, motor function, balance function, ADL, emo-tion of all the patients were assessed before and 21 days after rehabilitation intervention. Results After treatment, the motor function, bal-ance function, ADL and emotion of each rehabilitation group were improved. The patients'rehabilitation scores in FMA, Fugl-Meyer balance function, MBI and HRSD got much better than that in control group. The difference was statistic significant. The effect of rehabilitation treat-ment did not show difference among rehabilitation groups. Conclusion Early rehabilitation treatment for acute stroke does benefit for stroke patients. Effect does not show vary with the start time of rehabilitation treatment within 8 days after acute stroke.

OBJECTIVETo study influence of serum containing Liuwei Dihuang decoction (see text) on proliferation and differentiation of osteoblast form neonatal SD rats cultured in vitro at different times and different stretch stress.

METHODSAfter osteoblast cultured for 24 hours in the serum containing Liuwei Dihuang decoction (see text) and serum in control group, the 0.5 Hz frequency, 6% and 12% stretch-stress were added. The MTT1 and the activity of ALP were measured at the 12th and 24th hours, and the data were analyzed.

RESULTS1. In the environment of stretch stress to the frequency of 0.5 Hz, and stretched for 24 hours, the osteoblast was stimulated under elongation rate of 6% and 12%; the proliferation and differentiation of osteoblast was more active under elongation rate of 12% than that of 6%. 2. There were no stimulating effects on osteoblast proliferation and differentiation of serum containing Liuwei Dihuiang decoction (see text) acted on osteoblast cells of SD rats cultured in vitro for a shot time.

CONCLUSIONStretch stress environment can enhance osteoblast proliferation and differentiation cultured in vitro.

Alkaline Phosphatase ; metabolism ; Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Male ; Osteoblasts ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Serum ; Stress, Mechanical

Australia ; Environmental Monitoring ; Humans ; Occupational Health ; Safety Management ; methods ; organization & administration

OBJECTIVETo establish a more stable method to isolate osteocytes in vitro, and then to find the differences with osteoblast biological characteristics.

METHODSOsteocytes and osteoblasts were isolated from the bone tissue of 3-day-old rats using sequential collagenase digestion. The cells were identified through cell morphology after 24 hours later. Alkaline phosphatase (ALP) kit was used to stain the first generation cells by Kaplow-way, the bone gla protein (BGP) of the cells were stained by immunocytochemitry. Measured ALP and computed its activity.

RESULTSOsteocytes and osteoblasts showed obviously differences in cell morphology. Osteocytes were star-shaped or dendrite-shaped within more dendrites, while osteoblasts were spindle-shaped with short dendrites. Osteocytes were negative for ALP, but osteoblasts were positive; Osteocytes were more positive for BGP, and osteoblasts were less positive. The secretion of ALP in osteocytes was lower than that of osteoblasts.

CONCLUSIONOsteocytes can be isolated and cultured in vitro. These characteristics of osteocytes are apparently difference with those of osteoblasts.

Alkaline Phosphatase ; analysis ; Animals ; Cell Separation ; methods ; Osteoblasts ; chemistry ; cytology ; Osteocalcin ; analysis ; Osteocytes ; chemistry ; cytology ; Rats ; Rats, Sprague-Dawley

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Carcinoembryonic Antigen ; metabolism ; Carcinoma, Neuroendocrine ; metabolism ; pathology ; surgery ; Chromogranins ; metabolism ; Female ; Follow-Up Studies ; Gastrointestinal Neoplasms ; metabolism ; pathology ; surgery ; Humans ; Ki-67 Antigen ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis ; Synapsins ; metabolism

OBJECTIVETo establish a chiro chromatography for studying the stereoselective metabolism of propranolol (PL) in S(9) incubates prepared from transgenic cell lines expressing human cytochrome P450.

METHODSThe concentration of each enantiomer in S(9) incubates was determined through precolumn derivatization with GITC, followed by RP-HPLC assay using S-(+)-propafenone as internal standard.

RESULTSBaseline separations among the diastereomers of S(-)-P, internal standard and R(+)-PL were achieved on Shimpack CLC C(18)ODS column, with UV detection and methanol:water:glacial acetic acid (67/33/0.05,v/v/v) as mobile phase. The assay was simple, accurate, precise and specific. The linear range was from 5 to 500 micromol/L for each enantiomer. The limit of quantitation (LOQ) for the method was 5 micromol/L for the S(-)-and R(+)-PL, respectively (n=5, RSD<10%). The analytical method afforded average recoveries of 98.7 and 98.1% for S(-)- and R(+)-PL, respectively. The reproducibility of the assay was good (RSD<10%). The time-dependent studies showed that PL had the stereoselectivity of S-(-)-isomer in metabolism via CYP2C18 and the stereoselectivity of R-(+)-isomer in metabolism via CYP2C9.

CONCLUSIONThe method allows to study of stereoselective metabolism of PL in vitro.

Chromatography, High Pressure Liquid ; Cytochrome P-450 Enzyme System ; genetics ; physiology ; Humans ; Propranolol ; analysis ; metabolism ; Reproducibility of Results ; Stereoisomerism ; Transgenes

Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P

Objective:To investigate the polymorphisms at HLA-A,-B,-DRB1 loci in Dalian Han population. Methods: A total of 10 000 unrelated marrow donors who live in Dalian were genotyped by SBT and SSO methods. Haplotype frequencies and linkage dis-equilibrium values were calculated by ARLEQUIN software,and DA genetic distances between populations were calculated by poptree2 software. Results: A total of 18 HLA-A alleles, 32 HLA-B alleles and 13 HLA-DRB1 alleles were found in Dalian Han population. HLA-A?02 (31. 65% ),B?40(14. 84% ) and DRB1?15(15. 82% ) occurred most frequently. A?30-B?13-DRB1?07 (4. 56% ) was determined to be the most common three-locus haplotype and the second predominant haplotype was A?02-B?46-DRB1?09 ( 2. 43% ) . A ?30-B ?13 ( 6. 00% ) and B ?13-DRB1 ?07 ( 59. 89% ) were the most common two-locus haplotypes. Moreover,A?33-B?58 and B?13-DRB1?07 were strongest haplotypes with the linkage disequilibria values 0. 336 6 and 0. 665 1,respectively. In China,the closest genetic distances were found with Heilongjiang (0. 001) followed by Jilin (0. 002) and Shandong (0. 002),the furthest was found with Taiwan (0. 047). Compared with other populations worldwide,the closest genetic distances were found with Thailand (0. 029) and Korea (0. 03),the furthest was found with Italy (0. 183). Conclusion: Dalian Han population had rich polymorphism at HLA-A,-B,-DRB1 loci,and the distribution of HLA-A,-B and-DRB1 was in line with the charac-teristics of the northern population.