In order to provide scientific basics for exploitation and sufficient application of Dendrobium officinale leaves resources, the phenol-sulfuric acid method was applied to determine the polysaccharide content. The monosaccharides were derivated by PMP and the derivatives were identified by HPLC-DAD-ESI-MS(n) and the contents of mannose and glucose were determined simultaneously. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2004A) was employed to generate the mean chromatogram and similarity analysis of the samples was carried out. The results demonstrated that polysaccharide content, monosaccharide compositions and composition ratio had an obvious difference between stems and leaves. The polysaccharide content of stems was higher than that of leaves. Monosaccharide composition in leaf was significantly different from that in stem. The polysaccharide from stems was composed of mannose and glucose, however the polysaccharide of leaves was acid heteropolysaccharide and was mainly composed of five monosaccharides, including mannose, galacturonic acid, glucose, galactose and arabinose. The similarity value of the 14 batches was above 0.9, indicating that similarity of fingerprints among different samples was high. The study can provide evidence for expanding the medicinal parts of D. officinale.
Chromatography, High Pressure Liquid ; Dendrobium ; chemistry ; Mass Spectrometry ; Plant Extracts ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Polysaccharides ; chemistry

OBJECTIVETo establish the method of specific chromatogram analysis of ether extract of Dendrobium officinale for identification of D. officinale.

METHODChromatographic separation was carried out at 30 degrees C on an Ultimate C18 column (4.6 mm x 250 mm, 5 microm) eluted with methanol and water containing 0.2% phosphoric acid in a gradient elution at a flow rate of 1.0 mL x min(-1). The detection wavelength was set at 280 nm. The similarity evaluation system for chromatographic fingerprint of NPC (National Pharmacopoeia Committee) was adopted to specific chromatogram construction.

RESULTThe HPLC specific chromatogram of D. officinale was constructed with 6 common specific chromatographic peaks including naringenin as a reference peak.

CONCLUSIONThe method shows good precision and repeatability of relative retention time. It can be used to identify D. officinale.

Chromatography, High Pressure Liquid ; Dendrobium ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ethers ; chemistry ; Reproducibility of Results

OBJECTIVETo explore the reason of postoperative axial pain (PAP) complication caused by unilaterally open-door cervical laminoplasty with Centerpiece mini-plate fixations for the treatment of multilevel cervical spondylotic myelopathy(CSM).

METHODSThe clinical data of 79 patients with CSM who underwent unilaterally open-door cervical laminoplasty from January 2010 to December 2013 were retrospectively analyzed. There were 45 males and 34 females, aged from 48 to 75 years old with an average of (58.7±4.4) years, complicated with ossified posterior longitudinal ligament(OPLL) of 42 cases. Courses of disease were from 2.1 to 3.9 years with an average of (3.0±0.4) years. Decompression segment occurred in C₃-C₆ of 31 cases, C₃-C₇ of 9 cases, C₄-C₇ of 39 cases. The condition of PAP was record. Cervical curvature index, cervical lordosis angle, the rate of cervical instability, the motion of flexion and extension between PAP group and non-PAP group were compared preoperatively. Multivariate non-linear regression analysis was used to verify relationship between aforementioned parameters and incidence of PAP. JOA score of preoperative, postoperative 6 months and initial onset of PAP, the improvement rate of JOA score and Odom criteria at final follow-up were used to evaluate curative efficacy.

RESULTSAll the patients were followed up from 26 to 44 months with an average of (36±9) months. Among them, 12 patients occurred PAP who receive the conservative treatment. The rate of preoperative cervical instablility of PAP group were higher than that of non-PAP group(<0.05). Preoperative cervical instability was the only independent risk factor in predicting occurrence of PAP. There were no significant differences in cervical curvature, cervical lordosis index, the motion of flexion and extension between PAP and non-PAP group before operation. There were no significant differences in the improvement of nerve function and clinical effect between PAP and non-PAP group after operation(>0.05).

CONCLUSIONSPreoperative cervical instability is prone to inducing the respectively intervertebral motion disorder and imbalance of stress redistribution, which results in PAP after cervical unilateral laminoplasty. Correct treatment of preoperative cervical instability is a key factor to prevent the occurrence of PAP after cervical laminoplasty, which would not affect long-term nerve functional recovery pronouncedly.

Epiberberine, one of the most important isoquinoline alkaloid in Coptidis Rhizoma, possesses extensive pharmacological activities. In this paper, the liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to study phase I and phase II metabolites. A Thermo HPLC system (including Surveyor AS, Surveyor LC Pump, Surveyor PDA. USA) was used. The cocktail probe drugs method was imposed to determine the content change of metoprolol, dapsone, phenacetin, chlorzoxazone and tolbutamide simultaneously for evaluating the activity of CYP2D6, CYP3A4, CYP1A2, CYP2E1 and CYP2C9 under different concentrations of epiberberine in rat liver microsomes. The result showed that epiberberine may have phase I and phase II metabolism in the rat liver and two metabolites in phase I and three metabolites in phase II are identified in the temperature incubation system of in vitro liver microsomes. Epiberberine showed significant inhibition on CYP2D6 with IC50 value of 35.22 μmol L(-1), but had no obvious inhibiting effect on the activities of CYP3A4, CYP1A2, CYP2E1 and CYP2C9. The results indicated that epiberberine may be caused drug interactions based on CYP2D6 enzyme. This study aims to provide a reliable experimental basis for its further research and development of epiberberine.
Animals ; Berberine ; analogs & derivatives ; chemistry ; metabolism ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP2D6 ; metabolism ; Cytochrome P-450 CYP2D6 Inhibitors ; chemistry ; metabolism ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Male ; Microsomes, Liver ; drug effects ; enzymology ; metabolism ; Molecular Structure ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

3,4-Methylenedioxymethamphetamine (MDMA) is a substituted amphetamine with stimulating and hallucinogenic properties. Since MDMA induces "ecstasy" it is extensively used as a "recreational" drug. It has been well established that MDMA is neurotoxic and can result in long-term degeneration of cerebral 5-hydroxytryptamine (5-HT) nerve terminals in many species. The present study was undertaken to investigate the long-term neurotoxic effects of MDMA on cortical and hippocampal structures, by repeatedly administering MDMA in short time. Male Wistar rats were randomly assigned to control group and MDMA-treated group. MDMA (10 mg/kg) was administered to rats of MDMA-treated group, once per hour, total 40 mg/kg; rats of control group were treated with the same volume of saline. Thirty-two weeks after administering MDMA, the expression of serotonin transporter (SERT) mRNA and diazepam binding inhibitor (DBI) mRNA was detected by in situ hybridization. The expression of glial fibrillary acidic protein (GFAP) was detected by immunohistochemistry, and the degeneration of nerve terminals was demonstrated by Bielschowsky and Glee Marsland silver staining. The results showed that the expression of SERT mRNA in hippocampus decreased by 31.96%, while expression of DBI mRNA in neocortex increased by 40.51%, compared with the control group (P<0.05). The expression of GFAP in the brain tissue increased (P<0.05), while significant reduction of the nerve terminals in neocortex was demonstrated by silver staining, compared with the control group. These results suggest that the neurotoxicity of MDMA results in sustained cortical and hippocampal structural changes, which in turn result in disorder of the brain functions.
Animals ; Cerebral Cortex ; pathology ; physiopathology ; Diazepam Binding Inhibitor ; genetics ; metabolism ; Hippocampus ; pathology ; physiopathology ; Male ; N-Methyl-3,4-methylenedioxyamphetamine ; toxicity ; Neurotoxicity Syndromes ; etiology ; pathology ; physiopathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Serotonin Plasma Membrane Transport Proteins ; genetics ; metabolism

To figure out the stability and intestinal bacteria metabolites of rats in vitro of astragaloside IV ( AST), this research was done to explore the stability of AST in the artificial gastric juice. artificial intestinal juice and rat liver homogenate and the metabolism in rat intestinal in vitro. HPLC was used to calculate the remaining rate of AST in biological samples by measuring the content of AST, while metabolites were determined by combining the methods of TLC, HPLC and LC-MS/MS. It turned out that AST was difficult to metabolize in the artificial gastric juice, artificial intestinal juice and rat liver. Also, the metabolic pathway of AST was stepped by deglycosylation. Firstly, AST was converted to its secondary etabolites (6-O-β-D-glucopyranosyl- cycloastragenol, CMG) by removal of xylose moiety at C-3, then transformed into cycloastragenol (CAG) after hydrolytic removal of the glucose moiety at C-6. All the results suggested that the metabolism of AST in vivo occurs mainly in the intestinal by hydrolysis of glycosyl. In conclusion, hydrolysis of intestinal flora is the main reason that AST metabolizes.
Animals ; Bacteria ; metabolism ; Chromatography, High Pressure Liquid ; Drug Stability ; Intestines ; microbiology ; Liver ; metabolism ; Rats ; Rats, Sprague-Dawley ; Saponins ; chemistry ; metabolism ; Tandem Mass Spectrometry ; Triterpenes ; chemistry ; metabolism

This study was purposed to summarize the clinical characteristics and laboratorial data of blastic plasmacytoid dendritic cell neoplasm (BPDCN) in pediatric patients in order to enhance understanding this disease in diagnosis and therapy. A rare case of BPDCN in children was enrolled in this study. The blood routine test, examination of bone marrow cell morphology, histopathology and immunophenotype of the skin lesions were performed and analysed, the single cell suspensions of the biopsied skin mass were detected by flow cytometry. The results showed that tumor cells expressed CD4, CD56, CD43 and CD123, while not expressed CD19, CD20, CD3, CD8, CD13, CD11b and myeloperoxidase (MPO). According to the clinical and laboratorial features and the results from histopathological and immunophenotype examinations, BPDCN was confirmed. It is concluded that BPDCN in children is an extremely rare hematopoietic malignancy with presenting a rapidly and fatally aggressive clinical course. The diagnosis of this disease is mainly based on the clinical presentations, pathologic and immunohistochemical features. BPDCN is a highly aggressive disease, its prognosis is very poor, its pathogenesis remans still unclear. A standard treatment protocol for BPDCN has not yet been established.
Adolescent ; Dendritic Cells ; Hematologic Neoplasms ; Humans ; Male ; Skin Neoplasms ; Waldenstrom Macroglobulinemia

OBJECTIVEThe presence of liver fibrosis in patients with beta-thalassemia major has been demonstrated to be an important negative prognostic factor. Identification of liver fibrosis in early stage would be of great value. Hyaluronic acid (HA), type III pre-collagen (PC III), collagen IV (C IV) and laminin (LN) as serum markers were widely used in the diagnosis of liver fibrosis in patients with chronic viral infections or alcoholic liver diseases. However, their values in thalassemic liver fibrosis have not been studied. This work was to determine the serum HA, PC III, C IV and LN levels in children with beta-thalassemia major and evaluate the diagnostic utility.

METHODSerum HA, PC III, C IV and LN in 49 hospitalized children with beta-thalassemia major (aged 1 - 15 years with the media age of 6.27 years) and 41 healthy children served as controls (aged 1 - 13 years with media age of 6.40 years) were detected by radioimmunoassay (RIA). Forty-five of 49 cases were performed percutaneous liver biopsies, and the histopathological fibrosis was compared with the four serum markers. The correlation and discriminate analysis were used.

RESULTSAll the serum levels of HA, PC III, C IV and LN in beta-thalassemia were significantly higher than those in controls (P < 0.01). In 36 of 45 cases, the histopathology showed liver fibrosis including stage I and stage II by biopsies with a positive rate of 80%. The serum levels of four markers increased successively with the aggravation of liver fibrosis from stage 0 to stage II, and significant correlation was observed between the level of HA or PC III and the stage of fibrosis (HA, r = 0.379, P = 0.017; PC III, r = 0.455, P = 0.04). While there was no difference between the level of C IV or LN and fibrosis (C IV, r = 0.312, P = 0.053; LN, r = 0.310, P = 0.055). Using discriminate analysis, the discriminate function of co-detection of the four markers for the diagnosis of fibrosis was 0.002 HA + 0.003 PC III + 0.002 C IV + 0.006 LN-1.859, which had a sensitivity of 93.88%, specificity of 68.29%, predictive value of positive test and negative test of 77.97% and 90.32%, respectively. Moreover, there was a significant correlation between the serum level of HA or PC III and the liver iron concentration (HA, r = 0.318, P = 0.035; PC III, r = 0.305, P = 0.044).

CONCLUSIONThe results suggest that, in beta-thalassemia major with chronic liver disease, HA and PC III showed more practical value in diagnosing liver fibrosis than the levels of C IV and LN. The combination of the four serum markers could improve the accuracy and reliability of the diagnosis. A validation study is necessary before introducing into the prediction function during the clinical practice.

Adolescent ; Biomarkers ; blood ; Child ; Child, Preschool ; Collagen Type III ; blood ; Collagen Type IV ; blood ; Female ; Humans ; Hyaluronic Acid ; blood ; Infant ; Laminin ; blood ; Liver Cirrhosis ; blood ; complications ; diagnosis ; Male ; Prognosis ; beta-Thalassemia ; blood ; complications ; pathology

OBJECTIVETo observe the influence of therapy with Chinese medicine Lirukang Granule (, LRKG) combined with psychological intervention on anxiety states and sex hormones in patients with cyclomastopathy and menoxenia.

METHODSA total of 470 subjects were randomly assigned to three groups by the net-central randomization system, the treatment group (161 patients, treated with LRKG and psychological intervention), the Chinese medicine group (157 patients, treated with LRKG), and the psychological intervention group (152 patients, treated with psychological intervention). The dose of LRKG was 12 g three times per day; psychological intervention included establishing relations, cognitive intervention and psychological persuasion, 30-40 min per session, once a week. The therapy duration for all groups was three months. The efficacy was compared and anxiety state/State-Trait Anxiety Invertory (STAI) scoring was measured before and after treatment. The serum estradiol (E2), progesterone (P), prolactin (PRL) and follicle stimulating hormone (FSH) levels of 60 patients selected randomly from each group during the luteal phase were measured before and after treatment, and a group of 20 healthy women were evaluated for comparison. A follow-up was arranged for one year after treatment.

RESULTSThirty subjects were lost to follow-up. (1) Comparison of efficacy: the markedly effective rate and the total effective rate of the treatment group were 86.67% (131/150) and 98.00% (147/150), respectively; of the Chinese medicine group, 64.58% (93/144) and 90.27% (130/144), respectively; and of the psychological intervention group, 0% (0/146) and 3.42% (5/146), respectively. The markedly effective rate and the total effective rate in the treatment group were significantly higher than those in the Chinese medicine and psychological intervention groups (P < 0.05). (2) Comparison of STAI scoring: STAI scoring was decreased dramatically in the treatment group after treatment compared with that of the Chinese medicine group (P < 0.01), but there was no significant difference compared with the psychological intervention group. (3) Comparison of levels of sex hormones: E2, P, PRL and FSH of the three patient groups were disordered before treatment, and significantly different from healthy women (P < 0.01). After treatment, the levels of P and FSH of the treatment group were significantly increased (P < 0.01), E2 and PRL were significantly reduced, which were also significantly decreased compared with the psychological intervention groups (P < 0.01). (4) FOLLOW-UP: the markedly effective rate and the total effective rate of the treatment group remained higher than those of the other two groups after one year of treatment (P < 0.05). (5) Adverse reactions: no obvious adverse reactions were found among the three groups.

CONCLUSIONSTherapy with Chinese medicine combined with psychological intervention was effective for short-term and long-term treatment of cyclomastopathy and menoxenia. The mechanism might be related to the regulation of sex hormones.

Adult ; Behavior Therapy ; methods ; Breast Diseases ; physiopathology ; therapy ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Menstrual Cycle ; Menstruation Disturbances ; physiopathology ; therapy ; Middle Aged ; Prospective Studies ; Psychotherapy ; methods ; Risk Assessment ; Treatment Outcome ; Young Adult

Aim To investigate the protective roles of sonic hedgehog( Shh) signaling pathway in hypoxia-in-duced DNA damage with the neonatal rat cardiomyo-cytes. Methods The hypoxia model on neonatal car-diomyocytes was established with one to two days old Sprague Dawley rats by deprivation of oxygen and glu-cose ( OGD) . After pretreated with Shh pathway ago-nist SAG1.3 or antagonist GANT61, the survival rates of cardiomyocytes were assayed by MTT after OGD 6 hours or 12 hours. The protein levels of Shh pathway, phosphorylated histone H2AX at serine 139 (γH2AX), phosphorylated ATM (p-ATM), phospho-rylated p53 ( p-p53 ) , cleaved-caspase-3, Bcl-2 and Bax were detected by Western blot. The γH2AX foci was detected by immunofluorescence. Results Com-pared to control group, the protein expression of γH2AX, p-ATM, cleaved-caspase-3, p-p53 in OGD cardiomyocytes significantly increased, and Bcl-2/Bax ratio proportionally decreased. Particularly, the ex-pression of γH2AX, p-ATM was highest at OGD 6 h, and then gradually declined after OGD 12 h. After SAG1.3 pretreatment, the expression of γH2AX, p-ATM, cleaved-caspase-3 and p-p53 dramatically de-creased and the Bcl2/Bax ratio increased in OGD 6 h or OGD 12 h cardiomyocytes. On the contrary, in GANT61 pretreatment group, the expression of γH2AX, p-ATM, cleaved-caspase-3 and p-p53 signifi-cantly increased and the Bcl-2/Bax ratio decreased compared to the OGD 6 h or OGD 12 h cardiomyo-cytes. Conclusion The activation of Shh pathway protects cardiomyocytes against hypoxia-induced apop-tosis through inhibition of DNA damage.