OBJECTIVETo investigate the effect of Smad7 on the expressions of collagen I and alpha-smooth muscle actin (alpha-SMA) in HSC-T6 cell line activated by transforming growth factor-beta1 (TGF-beta1).

METHODSHSC-T6 cells stably expressing M2-flag protein were selected after co-infection of the cells with pTRE-Smad7-M2-flag and pTet-on. The optimal dose of doxycycline for inducing Smad7 was determined, and the effects of Smad7 over-expression on the expressions of collagen I and alpha-SMA in the cells activated by TGF-beta1 and on Smad2/3 phosphorylation were evaluated using Western blotting.

RESULTSThe optimal dose of doxycycline for inducing Smad7 expression was 2 mg/L. Smad7 over-expression induced by doxycycline decreased the expressions of collagen I and alpha-SMA in HSC-T6 cells activated by TGF-beta1, and down-regulated the level of Smad2/3 phosphorylation.

CONCLUSIONSmad7 over-expression inhibits Smad2/3 phosphorylation, and decreases the expression of collagen I and alpha-SMA in HSC-T6 cells induced by TGF-beta1 to inhibit the progression of liver fibrosis.

Actins ; genetics ; metabolism ; Cells, Cultured ; Collagen Type I ; genetics ; metabolism ; Genetic Therapy ; Hepatocytes ; cytology ; metabolism ; Humans ; Liver Cirrhosis ; metabolism ; therapy ; Smad7 Protein ; pharmacology ; Transforming Growth Factor beta1 ; pharmacology

Evaluation of the permeability mainly focuses on intestinal absorption in biopharmaceutics classification system (BCS). It is more complicated that the absorption and metabolism under multicomponent environment in biopharmaceutics classification system of Chinese materia medica (CMMBCS) compared with single component environment, which needs suitable mathematical models to be described. Therefore, with full consideration of existing single component mathematical algorithm combining with the characteristics of intestinal absorption and metabolism, we explored and designed a new mathematical algorithm of intestinal absorption and metabolism of multicomponent drug. Then we put forward a new coefficient, P (influence), the relative change rate of the single component's intestinal absorption and metabolism under multicomponent environment compared with single component environment, which described the influences of intestinal absorption and metabolism of the component under multicomponent environment. Moreover, P (influence) highlights the distinctive characteristics of multicomponent drug's intestinal absorption and metabolism, and lays the foundation for the construction of CMMBCS.
Algorithms ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Humans ; Intestinal Absorption ; Intestines ; chemistry ; metabolism ; Models, Theoretical ; Solubility

In this article we built formula database of health food containing Gardeniae Fructus with Traditional Chinese Medicine Inheritance Support System (V2.0). And on this basis, use data mining method such as association rules of the software, to analyze commonly used formula raw materials or materials combination of formula containing Gardeniae Fructus and raw material application having assisted function formula to protect chemical liver injury. The result shows that of the 71 health food formulas containing Gardeniae Fructus, most used materials are Gardeniae Fructus, Lycii Fructus, Angelica Sinensis Radix, Poria and so on. Commonly used materials combination mostly are Gardeniae Fructus and Lycii Fructus, Gardeniae Fructus and Angelica Sinensis Radix, Gardeniae Fructus and Poria, Gardeniae Fructus and Paeonia. There are nearly 18 healthcare functions of the health food containing Gardeniae Fructus, and most of these are assisted functions to protect chemical liver injury, and then immune modulating function. Of 23 formulas containing Gardeniae Fructus having assisted function formula to protect chemical liver injury, Gardeniae Fructus usually combined with traditional Chinese medicine which nourishs blood and liver such as Pueraria, Lycii Fructus, Hawthorn, Paeonia and Turnjujube. Analyzing formula raw materials application of health food containing Gardeniae Fructus contributes a lot to the further development and utilization.
Chemical and Drug Induced Liver Injury ; drug therapy ; Data Mining ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Food, Organic ; Fruit ; chemistry ; Gardenia ; chemistry ; Medicine, Chinese Traditional ; methods

To research the effect of Ginseng Radix et Rhizoma and Aconiti Lateralis Radix Praeparata compatibility on cardiac toxicity in rats by UPLC-Q-TOF/MS, and explore the endogenous markers and molecule mechanism. Different compatibility of Shenfu decoction were given to male Wistar rats at dosage of 20 g · kg(-1) for 7 days, collected the serum, and analyze the endogenous metabolites effected by Shenfu formulation by principal component analysis and partial least-squares analysis. Results showed that content of glutathione, phosphatidylcholine and citric acid decreased in mixed-decoction group, while ascorbic acid, uric acid, D-galactose, tryptophan, L-phenylalanine increased. The results showed cardiac toxicity of Aconiti Lateralis Radix Praeparata in Shenfu mixed-decoction. Shenfu co-decoction group showed a similar or weaker trend compared with control group, but most of them do not have a statistically significant. The results indicated the scientific basis of Shenfu compatibility by comparison of co-decoction group with mixed-decoction group. Shenfu compatibility can reduce cardiac toxicity induced by Aconiti Lateralis Radix Praeparata, and citric acid, glutathione, phosphatidyl choline, uric acid might be regarded as potential markers of cardiotoxicity.
Animals ; Biomarkers ; Cardiotoxicity ; Drugs, Chinese Herbal ; toxicity ; Glutathione ; blood ; Least-Squares Analysis ; Male ; Metabolomics ; methods ; Principal Component Analysis ; Rats ; Rats, Wistar

OBJECTIVETo determine the effects of the autologous nucleus pulposus on the pain-related behaviors of hind paws in rats.

METHODSThe nucleus pulposus harvested from autologous coccygeal vertebra was applied beside the unilateral L4 and L5 nerve roots without compression. The mechanical withdrawal threshold of both paws were measured in different times after surgery. And hematoxylin and eosin (HE) staining was applied to observe the changes of nucleus pulposus and nerve roots.

RESULTSMechanical sensitivity of the operated side in paws obviously increased after application of autologous nucleus pulposus beside the lumbar nerve roots without compression. And HE staining showed obviously inflammatory changes in the nucleus pulposus and vacuolation in the nerve roots.

CONCLUSIONSInflammation resulted from nucleus pulposus may contribute to the development of mechanical hyperalgesia. The results suggest that in addition to mechanical compression, inflammation may be an important factor in the pathogenesis of sciatica.

Animals ; Disease Models, Animal ; Hyperalgesia ; etiology ; pathology ; physiopathology ; Intervertebral Disc Displacement ; complications ; pathology ; physiopathology ; Male ; Radiculopathy ; complications ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley

Objective To investigate the discrepancy of aldosterone synthesis process and potential regulation abnormality between aldosterone-producing adenoma(APA) and normal adrenal(NA) with microarray. Methods cRNA probes labelled with biotin were prepared from mRNA of APAs(APA group,n=10) or NAs(control group,n=7).The probes were hybridized with oligonucleotide microarray of target gene expression profile.Expression levels were read from the fluorescent intensity scanned.The difference of gene expression profile was analyzed by computer software.Differentially expressed genes were verified by real-time RT-PCR. Results Compared with control group,97 genes were up-regulated and 168 genes were down-regulated in APA group.In the genes related to steroid hormone synthesis,only CYP11B2 was significantly up-regulated.In the physiologic regulators of aldosterone synthesis,CYB5A,CYP17A1,DUSP1 and HMGCR were down-regulated,while RENBP and NR1H2 were up-regulated.As a key enzyme in the biosynthesis of cortisol,the expression of CYP17A1 gene was inhibited. Conclusion Among the aldosterone synthesis related enzyme and corresponding regulatory genes in APA,CYP11B2 may be a key synthetase,and the suppressed physiologic regulators of aldosterone synthesis may indicate the existence of neoplastic modulation.

OBJECTIVESTo determine the effect of destroying capsaicin-sensitive primary afferents (CSPA) fibers on paw withdrawal mechanical threshold (PWMT) induced by the direct compression of L5 nerve root with autologous disc.

METHODSThe procedure used autologous disc of the rats from the coccygeal intervertebral discs to apply direct pressure to the L5 dorsal root. PWMT was measured at the different time points post-surgery and pre-surgery. The changes in spatial expression pattern of c-fos protein in the spinal cord were also determined at 3 weeks when PWMT decreased to the peak.

RESULTSThe pretreatment with capsaicin produced a complete prevention of mechanical hyperalgesia induced by disc compression. The direct compression of L5 nerve root produced an obvious expression of fos-like immunoreactivity neurons in the dorsal horn of the spinal cord, which was significantly decreased by pretreatment with capsaicin.

CONCLUSIONSThe study shows that CSPA fibers, which mainly terminated in superficial layers of dorsal horn, may play a key role in mechanical hyperalgesia in the new sciatica model.

Afferent Pathways ; physiopathology ; Animals ; Capsaicin ; pharmacology ; Disease Models, Animal ; Hyperalgesia ; physiopathology ; Intervertebral Disc Displacement ; complications ; Male ; Pain Threshold ; drug effects ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sciatica ; etiology ; metabolism ; physiopathology ; Spinal Nerve Roots ; metabolism

To study the enzyme kinetics of schizandrin metabolism in different gender in rat liver microsomes, liver microsomes were prepared from male or female rats. Schizandrin was incubated with rat liver microsomes. Schizandrin and its metabolites were isolated and identified by HPLC-UV method. Vmax, Km and Cl(int) of schizandrin in male and female rat liver microsomes were (21.88 +/- 2.30) and (0.61 +/- 0.07) micromol x L(-1) x min(-1) x mg(-1) (protein), (389.00 +/- 46.26) and (72.64 +/- 13.61) micromol x L(-1), (0.0563 +/- 0.0007) and (0.0084 +/- 0.0008) min x mg(-1) (protein), respectively. The major metabolites of schizandrin in female and male rat liver microsomes were 7,8-dihydroxy-schizandrin (M1) and 7, 8-dihydroxy-2-demethyl schizandrin (M2b), respectively. Ketoconazole, quinidine, and orphenadrine had different level effects on schizandrin metabolism in both male and female rat liver microsomes, and cimetidine still had some inhibitory effect in male liver microsomes. CYP3A and CYP2C11 may be the main P450 enzymes in schizandrin metabolism and their difference in rat liver microsomes may be the main reason for the sex difference of metabolic enzyme kinetics and metabolites of schizandrin in rats.
Animals ; Chromatography, High Pressure Liquid ; Cimetidine ; pharmacology ; Cyclooctanes ; isolation & purification ; metabolism ; Enzyme Inhibitors ; pharmacology ; Female ; In Vitro Techniques ; Ketoconazole ; pharmacology ; Lignans ; isolation & purification ; metabolism ; Male ; Microsomes, Liver ; metabolism ; Orphenadrine ; pharmacology ; Plants, Medicinal ; chemistry ; Polycyclic Compounds ; isolation & purification ; metabolism ; Rats ; Rats, Sprague-Dawley ; Schisandra ; chemistry ; Sex Factors ; Spectrophotometry, Ultraviolet

OBJECTIVETo investigate the effect of prescription compatibility on the pharmacokinetics of components from Dachengqi Decoction (DCQD, ) in rats.

METHODSTwenty-four male rats were randomly and equally divided into the DCQD group, Dahuang (Radix et Rhizoma Rhei, Polygonaceae) group, Houpo (Magnolia officinalis Rehd., Magnoliaceae) group, and Zhishi (Fructus Aurantii Immaturus, Rutaceae) group. The blood samples were collected before dosing and subsequently at 10, 15, 20, 30, 45 min, 1, 2, 4, 8, and 12 h following gavage. The levels of aloe-emodin, rhein, emodin, chrysophanol, honokiol, magnolol, hesperidin, and naringin in rat serum were quantified using a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for pharmacokinetic study.

RESULTSThe area under the curve (AUC), mean retention time (MRT), the peak concentration (C(max)) of aloe-emodin, rhein, emodin, and chrysophanol in the DCQD group were significantly different compared with the Dahuang group (P <0.05, respectively). The mean plasma concentration, C(max), and the absorption of Dahuang's component in the DCQD group were obviously lower at each time point than those in the Dahuang group, while the elimination process of Dahuang's component was obviously delayed (P <0.05). Half-lives of aloe-emodin, chrysophanol, and rhein were also extended in the DCQD group (P <0.05, respectively). In the DCQD group, the mean plasma concentration, AUC, C(max) and absorption of honokiol, and magnolol were significantly lower (P <0.01, respectively) at each time point than those in the Houpo group, while the drug distribution half-life time (T(1/2α)), the drug eliminated half-life time (T(1/2β)), MRT, and time of peak concentration (T(max)) were significantly delayed (P <0.05, respectively). Pharmacokinetic parameters of hesperidin and naringin in the Zhishi group were not significantly different as compared with the DCQD group (P >0.05, respectively), while the MRT of naringin was significantly longer.

CONCLUSIONSThe compatibility in Chinese medicine could affect the drug's pharmacokinetics in DCQD, which proves that the prescription compatibility principle of Chinese medicine formulations has its own pharmacokinetic basis.

Administration, Oral ; Animals ; Anthraquinones ; administration & dosage ; blood ; pharmacokinetics ; Biphenyl Compounds ; administration & dosage ; blood ; pharmacokinetics ; Drug Incompatibility ; Emodin ; administration & dosage ; blood ; pharmacokinetics ; Flavanones ; administration & dosage ; blood ; pharmacokinetics ; Hesperidin ; administration & dosage ; blood ; pharmacokinetics ; Lignans ; administration & dosage ; blood ; pharmacokinetics ; Male ; Plant Extracts ; administration & dosage ; blood ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare the clinical outcomes of intrasacrospinal muscular approach and posterior midline approach in treating far lateral lumbar disc herniation.

METHODSThe clinical data of 32 patients with far lateral lumbar disc herniation underwent transforaminal lumbar interbody fusion from January 2004 to January 2011 were retrospectively analyzed. The patients were divided into intrasacrospinal muscular approach group (11 males and 6 females ) and posterior midline approach group (10 males and 5 females). All patients were followed up from 12 to 18 months with an average of 15.3 months. Operative time, blood loss, postoperative draining volume were recorded and pre-and post-operative visual analog scale (VAS) and Oswestry Disability Index (ODI) were compared between two groups.

RESULTSOperative time, blood loss, postoperative draining volume in intrasacrospinal muscular approach group was less than that of posterior midline approach group (P < 0.05). There was no significant difference in VAS at final follow-up between two groups (P > 0.05); and the mean ODI in intrasacrospinal muscular approach group was less than that of posterior midline approach group (P < 0.05).

CONCLUSIONFor the treatment of far lateral lumbar disc herniation, intrasacrospinal muscular approach has less injury for paraspinal muscle and more satisfactory clinical outcome and is better method than posterior midline approach.

Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Spinal Fusion ; methods ; Tomography, X-Ray Computed