HPLC Fingerprint Analysis of Benchmark Sample of Yanghetang / 中国实验方剂学杂志

ObjectiveTo establish a high performance liquid chromatography（HPLC） fingerprint of Yanghetang benchmark sample， and evaluate its quality with chemometric methods， so as to provide a reference for the quality control of this benchmark sample. MethodHPLC was used to establish the fingerprint of Yanghetang benchmark sample with ZORBAX SB-C18 column（4.6 mm×250 mm， 5 μm）， the mobile phase was consisted of acetonitrile（A） -0.05% phosphoric acid aqueous solution （containing 0.05% triethylamine solution）（B） for gradient elution（0-5 min， 2%-3%A； 5-15 min， 3%-5%A； 15-65 min， 5%-30%A； 65-90 min， 30%-70%A）， the flow rate was 1.0 mL·min-1， the column temperature was 35 ℃， and the detection wavelength was 210， 260 nm. Traditional Chinese Medicine（TCM） Chromatographic Fingerprint Similarity Evaluation System （2012 edition） combined with cluster analysis， principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA） were used to evaluate the quality differences between different batches of Yanghetang benchmark samples， and to find the main chemical components responsible for the quality differences. ResultHPLC fingerprint of Yanghetang benchmark sample was established， 13 common peaks were identified and attributed to each common peak， including peaks 2 and 8 from Rehmanniae Radix Praeparata， peaks 10 and 11 from Cinnamomi Cortex， peaks 1， 3-6 from fried Sinapis Semen， peak 13 from Ephedrae Herba， and peaks 7， 9， 12 from Glycyrrhizae Radix et Rhizoma. Eight of them were identified by comparing with control substance， which were 5-hydroxymethylfurfural（peak 2）， sinapine thiocyanate（peak 4）， glycyrrhizin（peak 7）， verbascoside（peak 8）， cinnamic acid（peak 10）， cinnamaldehyde（peak 11）， glycyrrhizic acid（peak 12） and ephedrine hydrochloride（peak 13）. The similarities of the HPLC fingerprints of 15 batches of Yanghetang benchmark samples with the control fingerprint were all greater than 0.80. The three chemometric methods could classify the samples into two categories. Eight differential components were screened out， among which 5-hydroxymethylfurfural， sinapine thiocyanate， verbascoside and ephedrine hydrochloride were identified. ConclusionThe established fingerprint analysis method is accurate， stable and reproducible， which basically reflects the overall chemical composition of Yanghetang benchmark sample， and can provide a basis for establishment of quality standards for compound preparations of this famous classical formula.

The physiological and biochemical responses of a medicinal plant (Salvia miltiorrhiza L.) to stress caused by various concentrations of NaCl.

Salvia miltiorrhiza, which is commonly known as Danshen, is a traditional Chinese herbal medicine. To illustrate its physiological and biochemical responses to salt stress and to evaluate the feasibility of cultivating this plant in saline coastal soils, a factorial experiment under hydroponic conditions was arranged on the basis of a completely randomised design with three replications. Five salinity treatments (0, 25, 50, 75 and 100 mM NaCl) were employed in this experiment. The results showed that salinity treatments of <100 mM NaCl did not affect the growth of Salvia miltiorrhiza in a morphological sense, but significantly inhibit the accumulation of dry matter. Salinity treatments significantly decreased the Chl-b content but caused a negligible change in the Chl-a content, leading to a conspicuous overall decrease in the T-Chl content. The Na(+) content significantly increased with increasing hydroponic salinity but the K(+) and Ca(2+) contents were reversed, indicating that a high level of external Na(+) resulted in a decrease in both K(+) and Ca(2+) concentrations in the organs of Salvia miltiorrhiza. Salt stress significantly decreased the superoxide dismutase (SOD) activity of Salvia miltiorrhiza leaves in comparison with that of the control. On the contrary, the catalase (CAT) activity in the leaves markedly increased with the increasing salinity of the hydroponic solution. Moreover, the soluble sugar and protein contents in Salvia miltiorrhiza leaves dramatically increased with the increasing salinity of the hydroponic solution. These results suggested that antioxidant enzymes and osmolytes are partially involved in the adaptive response to salt stress in Salvia miltiorrhiza, thereby maintaining better plant growth under saline conditions.

Study on establishment of golden hamster buccal squamous carcinoma model for and its biological characteristics / 重庆医学

Objective To establish the golden hamster model of buccal squamous carcinoma and observe its biological character-istics .Methods 50 golden hamster were randomly divided into two group :experiment group (n=40) and control group(n=10) . Buccal mucosa of golden hamster were daubed by exposure 4-nitroquinoline-1-oxide (4NQO) in experiment group and tap water in control group .HE staining and immunohistochemistry were used to observe the tissue sample on the 8th and 12th week .The tissue samples of golden hamster buccal-mucosa cancer were used for the in-vitro subculture .Then flat cloning formation rate ,expression of CK and Vim ,and cell karyotype were detected .Results The observations of cell morphology and biology showed that the tissue of buccal squamous carcinoma were conformed to the basic characteristics of squamous carcinoma cell in 26 golden hamster(84 .6% ) after 12 weeks .The positive rate of CK and Vim were 96 .0% by immunohistochemical staining .The Chromosomes were tetraploid karyotype .Conclusion We successfully established the golden hamster model of buccal squamous carcinoma by the daub method of 4NQO .

Mechanism of Acupotome Therapy on Knee Osteoarthritis(review) / 中国康复理论与实践

@#: Acupotome therapy exhibits good efficacy on knee osteoarthritis in clinic. The mechanism based on its dynamic equilibrium theory is that acupotome therapy can restore normal joint stress state by changing the imbalance of mechanical equilibrium around the joint. Furthermore, it can ease inflammation, relieve pain and repair tissue structure so as to achieve its therapeutic effect. This review proposes new thoughts for the etiopathogenisis, pathology and treatment of KOA.

Regulating effects of aquaporin 4 expression in LoVo cells by serum containing total anthraquinone in rheum / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the regulating effects of serum containing total anthraquinone in rheum on the expression of AQP4 in cultured LoVo cells in vitro.</p><p><b>METHOD</b>Sixteen SD rats were randomly divided into 4 groups respectively treated with 0, 1.4, 2.5, 4.5 g x kg(-1) x d(-1) of total anthraquinone in rheum by intragastric administration. After 7 days of treatment, the rats were anesthetized and celiotomized to prepare the serum containing total anthraquinone in rheum. LoVo cells cultured in vitro were treated with different concentrations of total anthraquinone in rheum for 24 h. The expression levels of protein and mRNA of AQP4 IN LoVo cells were decided by Western blot and semiquantive RT-PCR.</p><p><b>RESULT</b>Western blot and semiquantive RT-PCR showed that serum containing total anthraquinone in rheum prepared from rats treated with 4.5 g x kg(-1) x d(-1) of total anthraquinone in rheum could inhibit the expression of mRNA and protein levels of AQP 4 in LoVo cells (P<0.01).</p><p><b>CONCLUSION</b>Serum containing total anthraquinone in rheum can inhibit both the transcription and translation of AQP4 gene, indicating that the regulation mechanism of rhubarb on the expression of AQP4 can be related to the cathartic effect of rhubarb.</p>

Study of Knee Osteoarthritis Gait Index on Function Evaluation-Time Distribution at the Sole and the Vertical Rebound Force at Stand Phase / 中国康复理论与实践

@# Objective To investigate time distribution of pressure of footplate in each stage and vertical anti-force from ground of patients with knee osteoarthritis (KOA).Methods 40 KOA patients, 10 aged people, and 10 common saplings were selected to test time distribution of pressure of footplate in each stage and vertical anti-force from ground in period of monopodia support.Results To unilateral KOA patient, there was not obvious difference to be observed in initial contact phase and forefoot push off phase ( P>0.05), but there was a significant difference in forefoot contact phase and foot flat phase between health side and sick side ( P<0.001), and there was obvious difference decreased buffer time and buffer force coefficient between them ( P<0.01).Conclusion To KOA patients, the time decreases in weight-bearing phase and absorption shocks, and amortization coefficient also decreases. These indexes can be used as the walking index of functional assessment and the objective evidence of curative effect.

Significance of Magnetic Resonance Imaging for Early Diagnosis of Knee Ostarthritis(review) / 中国康复理论与实践

@#During the latest researches on knee ostarthritis,it is discovered that before the morphology changes of cartilage,there were changes of the content of proteoglycan and water and changes of alinement of surface layer collagen fibers in cartilage matrix.By the new technologies of DWI,proton density mapping image,23Na MRI,T2Mapping image and so on,not only the pathological changes of cartilage can be disclosed,the component changes of cartilage matrix can also be detected.Accompany with the progress in the technology of software and hardware,MRI will have more and more prospects in early diagnosis and therapeutic effect evaluation of knee ostarthritis.

Inhibitory Effect of Growth-inhibitor on Isochrysis galbana and Inhibition of Antioxidants on Growth-inhibitor Damage to Its Cells / 生物化学与生物物理进展

The inhibitory effects of growth-inhibitor on the growth of Isochrysis galbana, malondialdehyde (MDA) content and activities of antioxidant enzymes (superoxide dismutase and peroxidase) were studied. And the inhibition effects of 4 species of antioxidants on this damage to its cells were also investigated. The results showed that 0.10 mg/L of GI marked decreased cell densities, activities of superoxide dismutase (SOD) and peroxidase (POD), and MDA content increased. With increase of GI concentrations, cell densities and activities of SOD and POD decreased sharply, and MDA content increased obviously. When concentration of GI was 0.30 mg/L, the cell densities, activities of SOD and POD decreased to 0.05, 0.56 and 0.59 folds of the control, respectively, and MDA content increased to as higher as 2.2 folds of that of the control set. L-Ascorbic acid, citrazinic acid, lenediamiaetetraacetic acid disodium salt (Na2EDTA) and 3-tert-butyl-4-hydroxyanisole could decrease MDA content obviously, and marked increased cell densities and activities of SOD and POD. When 4 species of antioxidants were added to the medium, the cell densities, and activities of SOD and POD increased to as high as 1.38～1.90, 1.49～2.12 and 1.55～2.13 folds of that of the control set, respectively, and MDA content decreased by 57.7%～87.9%. It was indicated that the excess active oxygens were produced under growth-inhibitor stress and active oxygen participated in the damage of growth-inhibitor to Isochrysis galbana; the L-ascorbic acid, citrazinic acid, lenediamiaetetraacetic acid disodium salt and 3-tert-butyl-4-hydroxyanisole could effectively inhibit membrane lipid peroxidation and further alleviated algal damage.

Determination of icariin and adenosine in Cordyceps Vigorine Tablet by HPLC / 中成药

AIM: To establish HPLC for the determination of icariin and adenosine in Cordyceps Vigorine Tablet. METHODS:MetaChem Polaris ODS C18-A column(4.6 mm × 250 mm, 5 μm) was adopted; acetonitrile-water was the mobile phase; detection wavelength was set at 272 nm for icariin and at 260 nm for adenosine; column linearity within the range of 0. 374-2. 246 μg for icariin and 0. 348-1. 392 μg for adenosine, respectively. The average recovery and RSD were found to be (99.4 ± 1.84) % for icariin ( n = 6), (99.63 ± 1.29 ) % for adenosine,respectively. CONCLUSION: The method is convenient, fast, reliable to operate and suitable for the quality control of Cordyceps Vigorine Tablet.

Protective effects of luteolin-7-glucoside against liver injury caused by carbon tetrachloride in rats.

Ixeris chinensis (Thunb.) Nakai has been used as a Chinese folk medicine; the information on the physiological and biochemical functions of the compounds extracted from I. chinensis is still scanty. We investigated the effects of luteolin -7-glucoside (LUTG) isolated from I. chinensis against liver injury caused by carbon tetrachloride (CCl4). CCl4 significantly increased the enzyme activities of glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) in blood serum, as well as the level of malondialdehyde (MDA) and 8-hydroxydeoxyguanosine (8-OHdG) in liver tissue, and decreased the levels of reduced glutathione (GSH). Pretreatment with LUTG was not only able to suppress the elevation of GPT, GOT, MDA and 8-OHdG, and inhibit the reduction of GSH in a dose-dependent manner in vivo, but also reduce the damage of hepatocytes in vitro. On the other hand, we also found LUTG has strong antioxidant activity against reactive oxygen species (ROS) in vitro in a concentration-dependent manner. The hepatoprotective activity of LUTG was possibly due to its antioxidant properties, acting as scavengers of ROS. These results obtained in vivo and in vitro suggest that LUTG had protective effects against hepatic oxidative injury induced by chemicals. Further studies on the pharmaceutical functions and immunological responses of LUTG may help in the development of a clinical application.

Operative complications in tethered cord syndrome and their management / 中华外科杂志

<p><b>OBJECTIVES</b>To find out the common complications induced by the operation on patients with tethered cord syndrome (TCS) and to discuss the mechanism and the treatment of these complications.</p><p><b>METHODS</b>During 1993 and 2001, the spinal cords of 87 patients with TCS were explored and released operatively. The operative complications were analyzed retrospectively and the results of prevention and treatment of these complications were evaluated.</p><p><b>RESULTS</b>The operative complications in patients with TCS included headache (43.7%), hemorrhage (31.0%), lumbago (21.8%), retethering (18.4%), lumbar instability (12.7%), neural injury (8.1%) and CSF leakage (4.6%).</p><p><b>CONCLUSIONS</b>To reduce operative complications, the pathology of TCS should be investigated thoroughly before operation. The techniques of microsurgery and electrophysiology should be used properly and some complications were reversible after treatment. Post-operation rehabilitation training should be stressed.</p>