Ginseng is a traditional medicine in Asian countries. Ginsenoside has the main active ingredient, exhibit cardiovascular, tumor, and central nervous system activities. In particular, protopanaxatriol-type ginsenosides Rh1, exhibits anti-inflammatory, anti-allergic, and memory improvement activities. Ginsenoside Rh1 is only found in trace amounts in Panax ginseng, Panax pseudoginseng var. notoginseng, and Panax quinquefolius. Biotransformation of rare ginsenosides has become an effective way. In this paper, the research progress of transformation of ginsenoside saponins by biotransformation to produce rare ginsenoside Rh1 is reviewed, which provides a useful reference for the further development and preparation of ginsenoside Rh1.

Using a UPLC-MS/MS (MRM) and cocktail probe substrates method, the metabolic fingerprint of the compatibility of Radix Aconite (RA) and Radix Paeoniae Alba (RPA) and its effect on CYP450 enzymes were investigated. These main CYP isoforms include CYP 1A2, CYP 2C, CYP 2E1, CYP 2D and CYP 3A. Compared with the inhibition effect of RA decoctions on CYP450 isoforms, their co-decoctions of RA and RPA with different proportions can decrease RA' inhibition on CYP3A, CYP2D, CYP2C and CYP1A2, but can not reduce RA' effect on CYP2E1. The metabolic fingerprints of RA decoction and co-decoctions with different proportions of RPA in CYP450 of rat liver were analyzed by UPLC-MS. Compared with the metabolic fingerprints of RA decoction, the intensity of diester-diterpenoid aconitum alkaloids decreased significantly, while the intensity of monoester-diterpenoid alkaloids significantly increased in the metabolic fingerprints of co-decoctions of RA and RPA. The results suggest that RA coadministration with RPA increased the degradation of toxic alkaloid and show the effect of toxicity reducing and efficacy enhancing.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 Enzyme Inhibitors ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Liver ; drug effects ; enzymology ; Metabolome ; Paeonia ; chemistry ; Rats ; Tandem Mass Spectrometry

To investigate the effect of compound Chinese traditional medicine PC-SPES II I in inhibiting proliferation of human prostate cancer cell LNCaP based on the androgen receptor (AR) signaling pathway. The effect of PC-SPES II on LNCaP cell proliferation was detected by MTT assay. According to the findings, at the mass concentration of 180-1 440 mg x L(-1), PC-SPES II significantly inhibited the proliferation of LNCaP cells; the IC50 of PC-SPES II at 24 h and 48 h were 311.48, 199.01 mg x L(-1), respectively. The flow Cytometry detection showed 240 mg x L(-1) PC-SPES II arrested cells in G2/M phase, and an obvious apoptotic peak appeared before G0/G1 peak and rose over time. Meanwhile, Hoechst 33258 staining revealed apoptotic cellular morphology. Annexin V-FITC/PI staining manifested an increase in apoptotic cell ratio at the PC-SPES II concentration of 480 mg x L(-1) in a dose dependent manner. The prostate specific antigen (PSA) secretion of LNCaP cells was tested by PSA ELISA kit. Besides, compared with 25 mg x L(-1) Bic, 480 mg x L(-1) PC-SPES II significantly reduced the cell secretion of PSA. The AR and PSA mRNA and protein expressions were detected by qRT-PCR and Western blot. According to the results, after the induction of LNCaP cells with synthetic androgen 25 μg x L(-1) R1881, 240-480 mg x L(-1) PC-SPES II notably down-regulated the AR and PSA mRNA and protein expressions and inhibited the translocation of AR from cytoplasm to nucleus. In summary, PC-SPES II significantly can inhibit the in vitro proliferation of LNCaP cells and arrest cell cycle arrest in G2/M phase. Its mechanism may be associated with the down-regulation of the AR and PSA expressions and the inhibition of AR nuclear translocation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Prostate-Specific Antigen ; genetics ; metabolism ; Prostatic Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; Receptors, Androgen ; genetics ; metabolism ; Signal Transduction ; drug effects

Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.
Aconitine ; analogs & derivatives ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Enzyme Inhibitors ; pharmacology ; Ketoconazole ; pharmacology ; Male ; Metabolic Networks and Pathways ; Microsomes, Liver ; enzymology ; metabolism ; Quinine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfaphenazole ; pharmacology ; Tandem Mass Spectrometry

Using brain microdialysis and LC-MS/MS to detect acetylcholine in rat brain to investigate the effects of ligustrazine. A liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of acetylcholine in rat brain dialysate sampling by microdialysis. The results indicated that ligustrazine administration by subcutaneous injection significantly increased Ach release in rat medial prefrontal cortex and nucleus accumbens in a dose-related manner. The drug' s effect on Ach release in rat brain could be directly detected by microdialysis combined with HPLC-MS/MS and this method is selective and sensitive.
Acetylcholine ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Ligusticum ; chemistry ; Male ; Microdialysis ; Nucleus Accumbens ; metabolism ; Plants, Medicinal ; chemistry ; Prefrontal Cortex ; metabolism ; Pyrazines ; administration & dosage ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

Objective To report the clinical and myopathological features in a patient with common variable immunodeficiency (CVID) with myositis.Methods A 33 years old man suffered from recurrent respiratory infection with fever over 10 years.The symptoms improved after anti-infection therapy.At the same time he presented with fatigue.Two years ago he developed general muscle weakness,hypertrophy and myotonia,especially in the hands,neck and thighs.Genetic test for myotonic dystrophy protein kinase (DMPK) and zinc finger protein 9 (ZNF9) was performed.Laboratory tests,electromyography,muscle ultrasound and muscle biopsy were performed.In addition to standard histological and enzyme histochemical stainings,immunohistochemical method was used with primary antibodies of mouse anti human monoclonal antibodies including CD8 for T-lymphocytes,CD20 for B-lymphocytes,CD68 for macrophages and MHC-Ⅰ for muscle membrane.Results Electromyography revealed myogenic changes and abound with myotonic potentials.There was muscle hypertrophy in muscle ultrasound.Lung biopsy showed chronic inflammatory changes.Serum hypoimmunoglobulin and anemia were found.Muscle biopsy showed muscle fiber necrosis and regeneration with lymphocyte and macrophage infiltration.There were no gene mutations in DMPK and ZNF9 gene.Conclusion Muscle hypertrophy and myotonia appeared in CVID with myositis.

Objective To investigate the cognitive-communication ability of people aged 20-30 years in China. Methods 139 normal participants took part in the Cookie Theft test. 8 different indexes were used to analyze the language samples. These indexes were total words number, the percentage of incorrect statement, dysfluencies, providing structure support, repetition, content units, elaborations of content units and irrelevancies. Results The rater consistence reliability was 1, 0.89, 0.85, 0.93, 0.92, 0.98, 0.98, 0.62, respectively. Conclusion The norms have acceptable reliability.

OBJECTIVETo screen specific small interfering RNA (siRNA) targeting mouse BMP9 gene and identify its function in BNLCL.2 fetal liver cells and C3H10 cells.

METHODSFour pairs of double-stranded DNA fragments for silencing mouse BMP9 were annealed in vitro and cloned into pSOS-BMP9 vector with BMP9 gene to construct pSOS-simBMP9 plasmid. The 4 pSOS-simBMP9 plasmids were separately transfected in HEK293 cells via Lipofectamine, and the gene silencing efficiency was assessed by GFP detection. BNLCL.2 fetal liver cells were infected with the constructed adenovirus simBMP9s, and their BMP9 expression was detected with RT-PCR and Western blotting. C3H10 cells were co-infected with Ad-simBMP9 and Ad-BMP9, and the inhibitory effect on BMP9-induced osteoblasts was evaluated by alkaline phosphatase (ALP) activity.

RESULTSGFP expression in the two simBMP9 groups was significantly decreased in HEK293 cells, and the endogenous expression of BMP9 was reduced by 50%-70% by adenovirus-mediated simBMP9 in the fetal liver cells. ALP activity in C3H10 cells was significantly higher in BMP9 group than in the control group (P<0.01), while the activity of the two Ad-simBMP9-infected groups was significantly lower than that in Ad-BMP9-infected group (P<0.01).

CONCLUSIONTwo specific siRNA targeting mouse BMP9 gene have been obtained, which can effectively inhibit both endogenous and exogenous expressions of BMP9 to facilitate the study of the mechanisms of BMP9 in liver cell differentiation.

Animals ; Cell Differentiation ; Cell Line ; Fetus ; Growth Differentiation Factor 2 ; biosynthesis ; genetics ; Hepatocytes ; metabolism ; Humans ; Mice ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo investigate the effect and mechanism of valsartan and felodipine extended release tablets (Plendil) on a novel salt-sensitive hypertensive rat induced by sensory denervation.

METHODSNewborn Wistar rats were given 50 mg/kg capsaicin subcutaneously on the 1st and 2nd day of life. Control rats were treated with vehicle solution (10%ethanol, 10%Tween 80 in saline). After weanling period (3 weeks), male rats were divided into 5 groups and subject to the following treatment for 4 weeks: control + high salt diet (4%, CON-HS), capsaicin + normal salt diet (0.5%, CAP-NS), capsaicin + high salt diet (CAP-HS), capsaicin + high salt diet + Valsartan (30 mg/kg per day, by orally) (CAP-HS-VAL), capsaicin + high salt diet + Plendil (30 mg/kg per day, by orally) (CAP-HS-PLE). Tail-cuff systolic blood pressure, body weight, intralymphocytic [Ca(2+)](i), plasma calcitonin gene-related peptide concentration ([CGRP]), angiotensin II concentration ([AngII]) and 24 hour water intake, urinary volume, urinary Na(+) and K(+) concentrations were examined.

RESULTSTail-cuff systolic blood pressure and intralymphocytic [Ca(2+)](i) were lower in CAP-HS-VAL or CAP-HS-PLE group than those in CAP-HS group. Plasma [AngII] were higher in CAP-HS-VAL group than that in other groups. Tail-cuff systolic blood pressure were lower in CAP-HS-VAL group than that in CAP-HS-PLE group. Intralymphocytic [Ca(2+)](i) were lower in CAP-HS-PLE group than that in CAP-HS-VAL group. The 24 hour urine sodium excretion was higher in CAP-HS-PLE group than that in CAP-HS or CAP-HS-VAL group.

CONCLUSIONValsartan or Plendil could prevent the development of salt-sensitive hypertension induced by sensory denervation and the overloading of intracellular [Ca(2+)](i), which indicated that salt-sensitive hypertension induced by sensory nerve degeneration might be related to renin-angiotensin-aldosterone system (RAAS) and the over loading intracellular [Ca(2+)](i), and might be more closely to RAAS.

Animals ; Antihypertensive Agents ; therapeutic use ; Blood Pressure ; Felodipine ; therapeutic use ; Hypertension ; chemically induced ; drug therapy ; physiopathology ; Male ; Rats ; Rats, Wistar ; Sodium Chloride, Dietary ; Tetrazoles ; therapeutic use ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan

Effects of six kinds of Chinese herb extracts, including Folium Crataegi extract, Herba Epimedii extract, Folium Acanthopanacis Senticosi extract, Trifolium pratense L. extract, Folium Ginkgo extract and Radix Puerariae extract, on the activities of CYP450 isozymes (CYP1A2, CYP2C, CYP2E1, CYP2D, CYP3A) in rat hepatic microsomals were studied by using a UPLC-MS/MS (MRM) and cocktail probe substrates method. The results showed that effects of six kinds of Chinese herb extracts on each CYP450 isozyme activity were inhibitory. The IC50 of Folium Crataegi extract for the inhibition of rat microsomal CYP2D activity was only for 4.04 microg x mL(-1), which showed the highest inhibition; Trifolium pratense L. extract had strong inhibitory action to CYP2D, the IC50 value was 5.73 microg x mL(-1); Folium Crataegi extract also had strong inhibitory action on CYP2E1, the IC50 value was 10.91 microg x mL(-1). Furthermore, the IC50 of Folium Ginkgo extract for the inhibition of rat microsomal CYP3A, 2D, 2E1 activities were 45.12, 35.45 and 22.41 microg x mL(-1), respectively, and the IC50 of Folium Acanthopanacis Senticosi extract on the inhibition of rat microsomal CYP2E1 activity was 32.89 microg x mL(-1). In addition, mechanism of inhibition experimental results showed that the inhibiting abilities of Folium Crataegi extract and Radix Puerariae extract on each CYP450 isozyme increased with the increasing of the preincubation time, therefore, the inhibitory effects were a mechanism-based inhibition.
Animals ; Chromatography, High Pressure Liquid ; Crataegus ; chemistry ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2E1 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Eleutherococcus ; chemistry ; Epimedium ; chemistry ; Ginkgo biloba ; chemistry ; Inhibitory Concentration 50 ; Male ; Microsomes, Liver ; enzymology ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Trifolium ; chemistry