Chemical profiling and tissue distribution study of Jingyin Granules in rats using UHPLC-Q-Exactive Orbitrap HR-MS / 中国中药杂志

In this study, the chemical profiling of Jingyin Granules and the tissue distribution of nine major constituents in this Chinese medicine were performed after oral administration of Jingyin Granules to rats, by using UHPLC-Q-Exactive Orbitrap HR-MS. An Acquity UPLC BEH C_(18) chromatographic column(2.1 mm×100 mm, 1.7 μm) was used as solid phase, while the mobile phase was methanol and 0.1% formic acid water for gradient elution. The major constituents in this Chinese medicine were quickly and accurately identified, via comparison with the retention times and MS/MS spectra of the standards. A total of 106 chemicals were identified from Jingyin Granules, including 24 kinds of organic acids, 47 kinds of flavonoids, 10 kinds of iridoids, and 21 kinds of saponins and 4 kinds of other compounds. After oral administered Jingyin Granules to rats, 48, 30, 25, 23, 45, 34, 39, 26, 19 prototype compounds were identified in serum, heart, liver, spleen, lung, kidney, brain, fat, and testicles, respectively. Meanwhile, an LC-MS based analytical method was established for simultaneous determination of chlorogenic acid, swertiamarin, caffeic acid, sweroside, liquiritin, prim-O-glucosylcimifugin, arctiin, 5-O-methylvisammioside and arctigenin in biological samples. The tissue distribution(serum, liver and lung) of these nine aim constituents in rats after oral administration of Jingyin Granules were investigated. It was found that these nine constituents could be quickly absorbed into circulation system and then distributed to liver and lung tissues. Except arctigenin, the exposure of other eight aim constituents to serum and lung was peaked at 1 h. At 1 h, the exposure of these components to lung tissue were ranked as follows: swertiamarin [(75 191.0±3 483.21) ng·g~(-1)]>arctiin [(2 716.5±36.06) ng·g~(-1)]>5-O-methylvisammioside [(585.1±0.71) ng·g~(-1)]>arctigenin [(437.45±3.18) ng·g~(-1)]>chlorogenic acid [(308.1±5.66) ng·g~(-1)]>prim-O-glucosylcimifugin [(211.35±2.19) ng·g~(-1)]>sweroside [(184.3±9.05) ng·g~(-1)]>caffeic acid [(175.95±2.05) ng·g~(-1)]>liquiritin [(174.78±153.34) ng·g~(-1)]. In summary, an UHPLC-Q-Exactive Orbitrap HR-MS method has been established for rapid and accurate identification of the constituents in Jingyin Granules, while the tissue distribution of nine major absorpted constituents were investigated in rats following oral administration of Jingyin Granules. These findings provided key information and guidance for further studies on pharmacodynamic substances and clinical applications of Jingyin Granules.

Progress on magnetic resonance imaging for evaluating the articular cartilage of the knee joint / 中国骨伤

The reasonable selection of magnetic resonance imaging(MRI) scan sequence and parameters is very important for the objective evaluation of the results of clinical study and high quality imaging. The semi quantitative scoring system of total knee joint including Whole Organ Magnetic Resonance Imaging Score, Boston Leeds Osteoarthritis Knee Score, MRI Osteoarthritis Knee Score, Cartilage Repair Osteoarthritis Knee Score and so on. They can fully evaluate the imaging changes of various organs during the development of knee osteoarthritis. With the continuous development of MRI technology, the morphological and physiological changes of articular cartilage can be quantitatively assessed. T2 mapping, Diffusion Weighted Imaging, and delayed Gadolinium-Enhanced MRI of Cartilage can be quantitatively monitoring changes in cartilage matrix components. These quantitative and semi quantitative evaluation techniques are helpful to detect OA in its early stage, guide clinical early intervention, and also provide the possibility for the accurate evaluation of the therapeutic effect.

Experimental study on topical treatment of diabetic skin ulcers with yi medicine "yi bu a jie" extract / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the role and mechanism of the Yi medicine, Yi Bu A Jie () extract, in topical treatment of diabetic skin ulcers, with a view to finding a breakthrough natural drug for the prevention and treatment of diabetic skin ulcers.</p><p><b>METHODS</b>A model of diabetic skin ulcers in Kunming mice was developed. Yi Bu A Jie was extracted in a Soxhlet extractor. Two different concentrations of the extract (0.005 mg/mL and 0.01 mg/mL) were applied to the wound of diabetic skin ulcers once every 3 days, and local skin appearance and histopathological changes were observed.</p><p><b>RESULTS</b>The shortest healing time was 25.25±2.06 day with a low concentration (P=0.0037 compared with the high concentration group, 33.14±2.21 day; P=0.0082 compared with control group, 28.21±2.14 days). The longest healing time was in the high concentration group (P=0.0025 compared with the control group). In both groups, a large number of inflammatory neutrophil cells were exuded during the experimental period. In the low concentration group, capillary-rich granulation tissue and actively growing fibroblasts appeared in the wound, while there was much necrotic tissue in the high concentration group.</p><p><b>CONCLUSION</b>Yi Bu A Jie extract has an inhibitory effect on diabetic skin ulcers in mice, and the low concentration is more suitable.</p>

Experimental study on the mechanism of icariin improving human osteoblasts proliferation and the expression of OPG protein / 中国骨伤

<p><b>OBJECTIVE</b>To establish the human osteoblasts culture system in vitro, observe the effects of icariin on human osteoblasts proliferation and expression of OPG protein, and to explore the mechanism of promoting bone formation about human osteoblast in icariin.</p><p><b>METHODS</b>The femoral cancellous bone pieces were obtained from the operation. The enzyme digestion method was used for culturing. The third passage of human osteoblast was taken for experiments. The cells were divided into four groups, the control group was treated with 15% NCS-DMEM-F12 (1:1), the experimental groups were respectively treated with 10(-6), 10(-8), 10(-10) mol/L icariin. The MTT method was used to observe the proliferation of human osteoblast on 1, 3, 5, 7, 9 d; in 8, 10, 12 d, western blot was used to determine the expression of OPG protein on human osteoblast.</p><p><b>RESULTS</b>1) Results of MTT: the icariin promoted the proliferation of human osteoblast. There was a concentration-response relation,while with the concentration of icariin increased, the ability was more obvious. There was statistically difference between 10(-6) mol/L icariin group (0.402 +/- 0.033) and the control group (0.268 +/- 0.031) (P<0.05). In the timely research, as the time prolong, the number of human osteoblast were more. At the fifth day, the human osteoblast entered rapid growth period, and access the growth platform stage; the icariin began to promote the proliferation of human osteoblast from the fifth day, which almost maintained to the 7th day and the 9th day, and most obvious in the 9th day. There was statistically difference between 10(-6) mol/L icariin group (0.402 +/- 0.033) and the control group (0.268 +/- 0.031) at the 9th day. (The results of OPG protein expression: in the control group, the expression of OPG protein was detected at the 8th day (1.01 +/- 0.08), and reached the expression peak (1.80 +/- 0.10), there was statistically different (P<0.05). In the different days and different concentration icariin groups, the expressions of OPG protein were all inferior to the control group. While the concentration decreasing, the expression was less. There were statistically difference (P<0.05). At the day 12, there was no significant difference of OPG protein expression between the 10(-6) mol/L icariin group and the 10(-8) mol/L icariin group (P>0.05).</p><p><b>CONCLUSION</b>The effect of icariin promoting the proliferation of human osteoblast maybe is one of the mechanisms of improving the bone formation on human osteoblast.</p>

Influence of Migu capsule and strengthening spleen prescription on the expression of small intestine VDR mRNA in calf muscle on ovariectomized rats / 中国骨伤

<p><b>OBJECTIVE</b>To observe the effect of Migu capsule and Strengthening Spleen prescriptions on the expression of vitamin D receptor on small intestine and calf muscle of the rat, while observing whether the Chinese medicine complex prescriptions of different effect such as invigorating the kidney and strengthening the spleen had selectivity to expression of the VDR mRNA.</p><p><b>METHODS</b>Copy the model of osteoporosis by ovariectomy operation, the rats were randomly divided into four groups, pseudo-resection group, model group, Migu capsule group and strengthening Spleen prescriptions group. Drug delivery 12 weeks later from operation, stomach lavage last for 12 weeks. Then, to observe the effect of medicine on the rats' bone mineral density, uterus weight, calf muscle/weight, and the VDR mRNA in small intestine and calf muscle through RT-PCR.</p><p><b>RESULTS</b>Migu capsule enhanced bone mineral density (P<0.05), raised calf muscle/weight (P<0.05), up-regulated expression of calf muscle VDR mRNA in the small intestine (P<0.01). Strengthening Spleen prescriptions remained bone mineral density (P>0.05), up-regulated expression of calf muscle VDR mRNA (P<0.01), there was no obvious difference in uterus weight in Migu capsule group and Strengthening Spleen prescription group.</p><p><b>CONCLUSION</b>Migu capsule and Strengthening Spleen prescriptions can cure osteoporosis by improving the expression of calf muscle VDR mRNA in the small intestine.</p>