[Effect of electroacupuncture on expression of heat shock protein 70 in mice with amyotrophic lateral sclerosis].

OBJECTIVE: To observe the effects of electroacupuncture (EA) on the body weight, disease progression and the expression of heat shock protein 70 (HSP70) in lumbar spinal cord of amyotrophic lateral sclerosis (ALS) mice, so as to explore the mechanism of EA on treating ALS. METHODS: Eighteen ALS transgenic SOD1G93A mice were randomly divided into model, EA and medication groups, with 6 mice in each group, and six C57BL/6J mice were used as the normal group. Mice in the EA group received EA at "Quchi"(LI11)-"Hegu"(LI4), "Zusanli"(ST36)- "Sanyinjiao"(SP6), 30 min/time, 5 times/week, for 8 weeks.Mice in the medication group were given riluzole solution (7.6 mg·kg-1·d-1) by gavage for 8 weeks. The body weight of the mice was recorded and the motor function of the hind limbs was evaluated by the neurological function scoring stan-dard of the ALS Therapeutic Development Institute. The expression of HSP70 in lumbar spinal cord was detected by Western blot and immunohistochemistry, respectively. RESULTS: Compared with the normal group, the body weight and the expression of HSP70 in the model group decreased significantly (P<0.05), while no significant difference in the body weight was found among other groups(P>0.05). After intervention and in comparison with the model group, the disease onset time and paralysis time of the EA group and the medication group were significantly delayed (P<0.05, P<0.01), the expression of HSP70 in the EA group and the medicine group was significantly increased (P<0.05, P<0.01).But there was no significant difference in the survival time among all groups(P＞0.05). The disease onset time of the EA group was shorter than that in the medication group (P<0.05). CONCLUSION: EA can increase the expression of HSP70 in lumbar spinal cord, thereby delaying the progression of ALS.

Exploring lipid biomarkers of coronary heart disease for elucidating the biological effects of gelanxinning capsule by lipidomics method based on LC-MS.

High-throughput lipidomics technology was used to explore the potential therapeutic targets and mechanism of action of gelanxinning capsule on rat model with coronary heart disease (CHD). This study attempts to provide a novel method to interpret the molecular mechanism of traditional medicine. The lipid markers of CHD were determined by full-scan analysis based on ultra-performance liquid chromatography-high-definition mass spectrometry. Then, the metabolic changes associated with gelanxinning capsule treatment via the modulation of lipid biomarkers and pathway in rats were characterized. After gelanxinning treatment, the metabolic profile tended to recover compared with the model group. A total of 26 potential biomarkers were identified to represent the disorders of lipid metabolism in CHD animal model, of which 19 were regulated by gelanxinning capsule administration, and four metabolic pathways such as glycerophospholipid metabolism, sphingolipid metabolism, glycosylphosphatidylinositol-anchor biosynthesis, and glycerolipid metabolism were involved. From the pathway analysis, it was found that glycerophospholipid metabolism and sphingolipid metabolism with significant differences have the potential to be regarded as new targets for the treatment of CHD. Gelanxinning capsule with its good therapeutic effect protects against CHD by regulating lipid biomarkers and pathway from lipidomics-guided biochemical analysis.

Metabolomics coupled with pattern recognition and pathway analysis on potential biomarkers in liver injury and hepatoprotective effects of yinchenhao.

Metabolomics can provide an opportunity to develop the systematic analysis of the metabolites in biological samples and has been increasingly applied to discovering and identifying biomarkers and perturbed pathways. It enables us to better understand the metabolic pathways which can clarify the mechanism of traditional Chinese medicines (TCM). Yinchenhao (YCH, Artemisia annua L), a famous TCM plant, has been used clinically for more than a thousand years to relieve liver diseases in Asia, and its mechanisms are not still completely clear. Here, metabolomic techniques may provide additional insight, and our investigation was designed to assess the effects and possible mechanisms of YCH on α-naphthylisothiocyanate (ANIT)-induced liver injury. Metabolite profiling was performed by ultra-performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry (UPLC/ESI-Q-TOF/MS) combined with pathway analysis and pattern recognition approaches including independent component analysis (ICA) and partial least squares-discriminant analysis (PLS-DA). Biochemistry test was also performed for the liver tissue and plasma samples. The changes in metabolic profiling were restored to their baseline values after YCH treatment according to the ICA score plots. Of note, YCH has a potential pharmacological effect through regulating multiple perturbed pathways to normal state, correlating well to the assessment of biochemistry test. Five different potential biomarkers in the positive mode contributing to the treatment of YCH were discovered. Pathway analysis showed that these metabolites were associated with perturbations in pyrimidine metabolism, primary bile acid biosynthesis, and propanoate metabolism, which may be helpful to further understand the action mechanisms of YCH. It showed that changed biomarkers and pathways may provide evidence to insight into drug action mechanisms and drug discovery.

[Analysis of syndrome discipline of generalized anxiety disorder using data mining techniques].

OBJECTIVE: To study the use of data mining techniques in analyzing the syndrome discipline of generalized anxiety disorder (GAD). METHODS: From August 1, 2009 to July 31, 2010, 705 patients with GAD in 10 hospitals of Beijing were investigated over one year. Data mining techniques, such as Bayes net and cluster analysis, were used to analyze the syndrome discipline of GAD. RESULTS: A total of 61 symptoms of GAD were screened out. By using Bayes net, nine syndromes of GAD were abstracted based on the symptoms. Eight syndromes were abstracted by cluster analysis. After screening for duplicate syndromes and combining the experts' experience and traditional Chinese medicine theory, six syndromes of GAD were defined. These included depressed liver qi transforming into fire, phlegm-heat harassing the heart, liver depression and spleen deficiency, heart-kidney non-interaction, dual deficiency of the heart and spleen, and kidney deficiency and liver yang hyperactivity. Based on the results, the draft of Syndrome Diagnostic Criteria for Generalized Anxiety Disorder was developed. CONCLUSION: Data mining techniques such as Bayes net and cluster analysis have certain future potential for establishing syndrome models and analyzing syndrome discipline, thus they are suitable for the research of syndrome differentiation.

Evaluation study on urine metabolomics in yinhuang rat model induced by triplet factors of rhubarb, ethanol, and α-nephthylisothiolyanate.

OBJECTIVE: To ascertain the biomarkers capable to characterize the animal composite model of Chinese medicine (CM) yinhuang syndrome induced by triplet factors of rhubarb, ethanol, and α-nephthylisothiolyanate (abbreviated as R, E, and A below) through metabolomic study and to evaluate the established model by means of studying the sources of markers based on the changes of metabolites produced from various combinations of the three modeling drugs. METHODS: Eighty Wistar rats allocated equally in eight groups (A-H) were treated with saline, R+E+A, R, E, A, R+E, R+A, and E+A, respectively. Rats' 12 h urine in the 14 successive experimental days were collected separately using metabolic cages and analyzed by ultra performance liquid chromatograph/time of flight mass spectrometer (UPLC/TOF-MS) to create the metabolic contour graph of urine in different groups for identifying the differences between them. The similarities and differences of metabolic network among various groups were represented from microcosmic viewpoint by pattern recognition method (principal component analysis). RESULTS: Controlled by group A, the landing points in principal component map of various groups were apparently assorted, especially obvious on the 14th day; 19 biomarkers, which capable to represent the genesis and development process of the yinhuang syndrome in the triplet factors-induced rat model, were identified. CONCLUSION: Metabolomic method is successfully used in evaluating the animal model of CM syndrome. Furthermore, according to the holistic view and substance changes in vivo, the influences of disease on organism were comprehensively analyzed, and the pathogenic mechanism of CM yinhuang syndrome was explored at the level of metabolomics in vivo as well.

Proteomic study of serum proteins in a type 2 diabetes mellitus rat model by Chinese traditional medicine Tianqi Jiangtang Capsule administration.

Proteomics technology was for the first time applied to investigate the changes of serum proteins levels in type 2 diabetes mellitus (T2DM) rat model after treated by Chinese traditional medicine Tianqi Jiangtang Capsule (ten normal Wistar rats, ten with T2DM and ten with T2DM administrated by Tianqi Jiangtang Capsule). In addition to two-dimensional polyacrylamide gel electrophoresis (2-DE), serum protein profiling in the three groups was further performed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-TOF/MS). 11 visualized spots were differentially regulated and identified as diabetes-associated proteins. All the samples in three groups were then analyzed by ELISA and estimated the 7 proteins which were found to vary. The distinct effect of T2DM induction on the pattern of rat serum includes the down-regulation of Apolipoprotein E, Apolipoprotein A-I, Ig gamma-2A chain C region, and up-regulation of Transthyretin (TTR), Haptoglobin (Hp), Serum amyloid P-componen (SAP), Prothrombin. The majority of those protein levels were interestingly restored to those of healthy rats after Tianqi Jiangtang Capsule treatment.

Two new monoterpenes from Sibiraea angustata.

Two novel monoterpenes, sibiscolacton (1) and sibiraic acid (2), were isolated from the aerial part of Sibiraea angustata RCHD: . along with seven known compounds, namely three phenylpropanoids (3-5), two flavonoids (6,7), one glucityl ferulate (8, sibirate), and a monoterpene glucoside (9, sibiskoside). The structures of 1 and 2 were established on the basis of spectroscopic and chemical data.

Comparative study on the protective effects of Yinchenhao Decoction against liver injury induced by alpha-naphthylisothiocyanate and carbon tetrachloride.

OBJECTIVE: To optimize the animal model of liver injury that can properly represent the pathological characteristics of dampness-heat jaundice syndrome of traditional Chinese medicine. METHODS: The liver injury in the model rat was induced by alpha-naphthylisothiocyanate (ANIT) and carbon tetrachloride (CCl(4) ) respectively, and the effects of Yinchenhao Decoction (, YCHD), a proved effective Chinese medical formula for treating the dampness-heat jaundice syndrome in clinic, on the two liver injury models were evaluated by analyzing the serum level of alanine aminotransferase (ALT), asparate aminotransferase (AST), alkaline phosphatase (ALP), malondialchehyche (MDA), total bilirubin (T-BIL), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) as well as the ratio of liver weight to body weight. The experimental data were analyzed by principal component analytical method of pattern recognition. RESULTS: The ratio of liver weight to body weight was significantly elevated in the ANIT and CCl(4) groups when compared with that in the normal control (P<0.01). The contents of ALT and T-BIL were significantly higher in the ANIT group than in the normal control (P<0.05,P<0.01), and the levels of AST, ALT and ALP were significantly elevated in CCl(4) group relative to those in the normal control P<0.01). In the YCHD group, the increase in AST, ALT and ALP levels was significantly reduced (P<0.05, P<0.01), but with no significant increase in serum T-BIL. In the CCl(4) intoxicated group, the MDA content was significantly increased and SOD, GSH-PX activities decreased significantly compared with those in the normal control group, respectively (P<0.01). The increase in MDA induced by CCl(4) was significantly reduced by YCHD P<0.05). CONCLUSION: YCHD showed significant effects on preventing liver injury progression induced by CCl(4), and the closest or most suitable animal model for damp-heat jaundice syndrome may be the one induced by CCl(4).

[Effect of geniposide on serum IL-1beta and TNF-alpha of rheumatoid arthritis rats].

OBJECTIVE: To study the effect of geniposide on serum IL-1beta and TNF-alpha levels of rheumatoid arthritis rats, as well as the mechanism of this drug. METHOD: To establish an experimental rat model of type II collagen-induced arthritic (CIA). The inhibitory effects on paw edema were observed, and serum IL-1beta and TNF-alpha levels were determined in experimental rats. RESULT: Compared with the model, geniposide delayed the starting time of right paw edema significantly, and the levels of serum IL-1beta and TNF-alpha were significantly decreased by geniposide at high dose or medium dose (P < 0.01). CONCLUSION: Geniposide can lower serum IL-1beta and TNF-alpha levels in rheumatoid arthritis rats. The effect may be close related to inhibitory development of rheumatoid arthritis by the agent.

[Study on the collecting method of constituents absorbed into the blood from anti-V capsule].

OBJECTIVE: Aim To establish the condition and parameters for collecting the effective constituents of Anti-V capsule. METHOD: Based on HPLC, using chlorogenic acid and baicalin which are the main constituents absorbed in the blood as the marker substance, the best technical process of macropore absorbed resin was evaluated by the quantity of two compounds in the collected part. RESULT: After the abstraction of the capsule, the collection method of NKA-2 macropore absorbed resin as sorbent and 5 bed volume 50% ethanol as mobile phase was definited as the best craftwork. In the collected part, the content of chlorogenic acid and baicalin was five times than in the capsule. CONCLUSION: In guide of serum pharmacochemistry, macropore absorbed resin is the effective method to collect the constituents absorbed in blood of Anti-V capsule. At the same time, this method provides a way to research other Compound medicine.