The study is to establish an HPLC method using fluorescence detector for the determination of doxazosin enantiomers and investigate their chiral inversion in vitro and in vivo. Ultron ES-OVM was taken as the chiral chromatographic column, and the column temperature was 30 degrees C. Isocratic elution using a mobile phase of phosphate buffer-acetonitrile (85 : 15, v/v) at a flow rate of 0.8 mL x min(-1) was done. The fluorescence detection was set at lambda(Ex) = 255 nm and lambda(Em) = 385 nm. Prazosin was used as the internal standard. (-) Doxazosin or (+) doxazosin added into rat plasma in vitro was determined after incubating in 37 degrees C water bath for 2, 5 and 10 days. (-) Doxazosin or (+) doxazosin was administered orally to the rats for one months. Plasma samples were taken at 8 h after the last administration. A good linear relationship was achieved when the concentration of doxazosin enantiomers was within the range of 4 - 2 000 ng x mL(-1). The average recovery for (-) doxazosin was 99.5% with RSD 3.6%, and for (+) doxazosin was 99.3% with RSD 4.3%. Chiral inversion was observed neither in vitro nor in vivo studies. The method is selective, accurate and reproducible, which is suitable for the detection of doxazosin enantiomers in rat plasma. The in vitro and in vivo studies indicate that chiral inversion occurs uneasily between (-) doxazosin and (+) doxazosin in the rat.

OBJECTIVEComparative study to the effect of Chinese herbal medicine on left ventricular remodeling in rats with left heart failure after myocardial infarction (MI).

METHODSRat's model of left heart failure after myocardial infarction was treated with injection for activating blood circulation (ABCI, consisted of R. Salviae miltiorrhizea; Rh. Ligusticum wallichii and F1. Carthamus tinctorius) and injection for replenishing Qi (RQI, consisted of R. Codonopsis Pilosulae and R. Astragalus membranaceus) respectively. The effect of treatment were evaluated by observing and comparing the changes of heart morphological structure, collagen element, heart weight/body weight ratio (HW/BW), left intraventricular area (LVA), ratio of ventricular wall thinning in MI area and myocardial nuclei number (MNN) per square area.

RESULTSIn comparison with the model group, the reduction of collagen tissue around myocardial cells in living area of MI, HW/BW and LVA of ABCI and RQI group were lower, and MNN per square area was higher significantly (all P < 0.05).

CONCLUSIONBoth ABCI and RQI, though without positive myodynamia, showed certain inhibitory effect of left ventricular remodeling in rats with left heart failure after MI.

Animals ; Astragalus membranaceus ; Cardiac Output, Low ; pathology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Myocardial Infarction ; pathology ; physiopathology ; Phytotherapy ; Qi ; Rats ; Rats, Wistar ; Ventricular Remodeling ; drug effects

Objective To determine the activities of 131I for treating differentiated thyroid carcinoma with diffuse pulmonary metastases ( DTC-DPM ) from the perspective of internal radiation dosimetry.Methods According to Medical Internal Radiation Dosimetry (MIRD) schema, the activity constraint,from which the whole bdy retention at 48 h should not exceed 2.96 GBq (2.96 GBq rule), was converted to dose-rate constraint(DRC) to lungs at 48 h ( DRCLU ·48 h ) in 131I therapy for DTC-DPM. Based on the assumption of DRCLU·48 h at 48 h in lung, the fractions of whole body activities ( F48 ), the effective half times of 131I in lungs ( TLL ) and the remainder of body ( TRB ) were 0.6-0.9, 20- 120 h, and 10- 20 h, respectively. The maximum safe activities of 131I for different human phantoms from the Organ Level Internal Dose Assessment (OLINDA) software were calculated. Results According to MIRD schema and 2.96 GBq rule, DRCLU ·48 h should not exceed 46.4 mGy/h in 131I therapy for DTC-DPM. Depending on varying F48 h,TLL and TRB, the maximum safe activities of 131I were 6.77-81.36, 5.29-56.20, 5.08-55.19 and 3.87-40. 52 GBq for the male adult, female adult, 15-year-old, and 10-year-old patients with DTC-DPM, respec tively. Conclusion Dosimetry-guided 131I therapy for DTC-DPM considers adequately the differences of 131I kinetics in individual patients and can adjust administered activities of 131I on the precondition of avoiding radiological pneumonitis and pulmonary fibrosis.

Abstract: Objective　To investigate a poisoning incident caused by eating eight treasure congee, and establish liquid chromatography (LC)-mass spectrometry (MS)/MS screening method of 28 alkaloids to provide references for disposal of similar poisoning incidents. Methods　LC-MS/MS was used for screening 28 alkaloids in the urine, eight treasure congee and food raw material, and the detected alkaloids were quantified. Samples were extracted with 0.4% formic acid aqueous solution and separated by a Acquity UPLC BEH C18 column (1.7 μm, 100 × 2.1 mm). Acetonitrile-0.2% formic acid aqueous solution was used as the mobile phase and gradient elution was adopted. The ionization mode was electrospray positive ionization mode, and the detection method was multi-reaction monitoring (MRM). Analytes were quantified with the external standard method. Results　In the concentration range of 0-100 ng/mL, the linear correlation coefficient r were greater than 0.999 for 28 alkaloids. The recovery of 28 alkaloids in urine sample ranged from 63.0% to 105.0%, and the relative standard deviations (RSDs) were between 5.8% and 8.6%. The recovery of 28 alkaloids in eight treasure congee sample ranged from 72.0% to 109.0%, and the RSDs were between 6.3% and 9.7%. The recovery of 28 alkaloids in semen sesami nigrum sample ranged from 60.0% to 95.0%, and the RSDs were between 4.8% and 8.2%. Hyoscyamine (2 380.0 ng/mL), scopliamine (3.6 ng/mL) and rac-anisodamine (4.7 ng/mL) were detected in the patient's urine. Hyoscyamine (63.3 μg/g), scopliamine (5.7 μg/g) and rac-anisodamine (2.1 μg/g) were detected in eight treasure congee. Hyoscyamine (901.0 μg/g), scopliamine (80.0 μg/g) and rac-anisodamine (30.1 μg/g) were detected in the seed of Datura stramonium L. The ratio of scopliamine and hyoscyamine in the seed of D. stramonium was 1∶11, which complies with the characteristics of D. stramonium L. In urine sample, the proportion of scopliamine and rac-anisodamine was 0.15% and 0.20%, and hyoscyamine accounted for 99.65%. Conclusion　Seed morphology, the content range and proportion of three alkaloids are all in accord with the characteristics of D. stramonium. Combined with the clinical symptoms of atropine poisoning, it can be deduced that this incident is a family food poisoning caused by accidental consumption of seed of D. stramonium L. The method can provide technical support for the clinical diagnosis and treatment of alkaloid poisoning patients, and also provide a basis for emergency detection and disposal of alkaloid poisoning events.

In this study, we investigated the effect of Cigu Xiaozhi formula on HSC-T6 activity in hypoxic microenvironment based on network pharmacology and computer-aided drug design, and predicted and verified its possible targets and related signaling pathways. The potential active components and targets of Cigu Xiaozhi formula were screened by searching Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Encyclopaedia of Traditional Chinese Medicine (ETCM) and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) databases, and the liver fibrosis related targets retrieved from Gene Cards and Pharm GK database were integrated to obtain the potential targets of Cigu Xiaozhi formula in the treatment of liver fibrosis. GO enrichment analysis and KEGG signaling pathway enrichment analysis were performed on Omic Share platform, and Cytoscape software was used to construct the "potential active ingredient-key target-pathway" network. The active components and target proteins were subjected to molecular docking analysis by Auto Dock software. According to the results of molecular dynamics simulation and binding free energy calculation, the top 5 active components with degree were scored. The active components stigmasterol and β-sitosterol were subjected to molecular docking. CoCl₂ was used to induce HSC-T6 cells to construct hypoxia model in vitro. The cell viability was detected by CCK-8 assay, and the optimal time and concentration of hypoxia model of HSC-T6 cells was determined to be 100 µmol·L^-1 CoCl₂ for 24 h. Under hypoxia condition, HSC-T6 cells were activated, the wound healing rate was significantly increased, and the fluorescence signal of activation marker protein α-smooth muscle actin (α-SMA) was significantly enhanced. However, 6% drug-containing serum could inhibit the activation of HSC-T6 cells, and the wound healing rate was significantly decreased, and the fluorescence signal of α-SMA was significantly weakened. Further studies showed that the expressions of hypoxia-inducible factor-1α (HIF-1α), α-SMA and key proteins of Hedgehog (Hh) signaling pathway in HSC-T6 cells were up-regulated under hypoxia, while the expressions of HIF-1α, α-SMA, Patched-1 (Ptch-1) and glioma related oncogene homology-1 (Gli-1) were down-regulated in 6% drug-containing serum group, the YC-1 group and the cyclopamine group. These results indicated that HIF-1α and Hh signaling pathways were involved in the activation of HSC-T6 cells, and the traditional Chinese medicine Cigu Xiaozhi formula could inhibit the activation of HSC-T6 cells, and the mechanism may be related to the inhibition of HIF-1α expression and the blocking of Hh signaling pathway. In conclusion, Cigu Xiaozhi formula can inhibit the activation of HSC-T6 cells by directly acting on HIF-1α and Hh signaling pathway, and exert an anti-hepatic fibrosis effect. The animal experimental protocol has been reviewed and approved by Laboratory Animal Ethics Committee of Gansu University of Chinese Medicine, in compliance with the Institutional Animal Care Guidelines.

Objective To explore the polymorphism of HLA class I alleles of HIV-infected former plasma donors and to investigate its impact on HIV-1 viral load in central China.Methods 106 subjects chronically infected with HIV-1 were recruited and HLA class I alleles were genotyped with PCR-SSP assay.HLA class I genotypes and haplotypes were determined and their association with plasma viral loads were analyzed.Gag-specific CTL responses were detected by an IFN-λ EUSPOT assay by using overlapping peptides,and their association with plasma viral loads were also analyzed.Results Subjects homozygous at HLA class I locus had higher plasma viral loads(P=0.0098);HLA-A*30,-B*13,-Cw*06,-Cw*14 alleles and HLA-A*30/B*13/Cw*06 haplotype were associated with lower plasma viral loads(P=0.0004,0.0103,0.0058,0.0371 and 0.0006);an inverse correlation between p2p7p1p6-specific CTL responses and viral loads in subjects with HLA-A*30/B*13/Cw*06 haplotype as well as an inverse correlation between p17-specific CTL responses and viral loads in subjects with HLA-Cw*14 allele were observed.Conclusion HLA-A*30,-B*13,-Cw*06,-Cw*14 alleles and HLA-A*30/B*13/Cw*06 haplotype were associated with lower plasma viral loads and Gag-specific CTL responses restricted by these HLA alleles may contribute to the protection.

OBJECTIVETo establish methods for the determination of phillyrin and forsythoside A in Lianqiao Bingdu Qing capsule by RP-HPLC.

METHODThe determination of phillyrin was carried out on YWG-C18 column (4.6 mm x 250 mm, 10 microm), using acetonitrile-water (25:75) as mobile phase at a flow rate of 0.8 mL x min(-1) and detected at the wavelength 277 nm. The determination of forsythoside A was carried out with YWG-C18 column(4.6 mm x 250 mm,10 microm), using acetonitrile-water-Acetic acid (17:83:0.4) as mobile phase at a flow rate of 1.0 mL x min(-1) and detected at the wavelength 280 nm.

RESULTThe average recovery of phillyrin was 99.6%, RSD = 1.9% (n = 5). The average recovery of forsythoside A was 101.3%, RSD = 2.5% ( n = 5).

CONCLUSIONThe methods were simple and accurate and could be used to control the quality of the Lianqiao Bingdu Qing capsule.

Capsules ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Forsythia ; chemistry ; Fruit ; chemistry ; Glucosides ; analysis ; Glycosides ; analysis ; Plants, Medicinal ; chemistry

This study is to establish the fingerprint for Phyllanthus emblica and their tannin parts from different habitats by HPLC for its quality control. The determination was carried out on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column, with methanol-0.2% glacial acetic acid as mobile phase with gradient elution at a flow rate of 1 mL x min(-1). The temperature was maintained at 30 degrees C and the detected wavelength is 260 nm, Thirteen chromatographic peaks were extracted as the common peaks of the fingerprint of P. emblica, and eleven as the common peaks of P. emblica tannin parts, and five peaks were identified by comparing with referent samples. The fingerprints of 8 samples were compared and classified by similarity evaluation, cluster analysis and principal component analysis (PCA). The similarity degrees of eight P. emblica were between 0.763 and 0.993, while tannin parts were between 0.903 and 0.991. All the samples of P. emblica and their tannin parts were classified into 3 categories. The method was so highly reproducible, simple and reliable that it could provide basis for quality control and evaluation of P. emblica from different habitats.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Medicine, Tibetan Traditional ; Phyllanthus emblica ; chemistry ; classification ; Quality Control ; Tannins ; analysis ; Tibet

· AIM: To inspect and compare the functional vision of an aspheric intraocular lens (Tecnis) with those of conventional monofocal silicone and acrylic intraocular lens and multifocal intraocular lens (Array).· METHODS: The IOLs were tested in the eye model, which was designed to be optically equivalent to the theoretical eye model. The eye model is a combination of a spherical photographic lens with 35mm focal length ( IOL put in a water cell)and a charge coupled device (CCD) camera. The images constructed by the lenses are observed on a monitor of personal computer and the contrasts of the images are analyzed by using commercial image processing software.· RESULTS: The modulation transfer function of the eye model equals the scale produced by the theoretical eye model. The images constructed by changing the diameter of aperture stop and IOL.· CONCLUSION: The proposed eye model is useful for testing functional vision and for inspecting differences of intraocular lens.

Objective To investigate the influence of high intensity interval exercise (HIIT) on peroxidation and vascular endothelial function for experimental hyperhomocysteinemia (HHcy) rats. Methods Thirty five male rats were randomly divided into 4 groups. Control group (n=8) was given ordinary feed. High methionine group (n=27) was given 3% methionine on this basis, and divided into model group, folic acid group and HIIT+ folic acid group, with 9 rats per group for 16 weeks. Serum homocysteine (Hcy) , content of plasma malondialdehyde (MDA) , hydroxyl radical (OH-), total antioxidant capacity (T-AOC), activity of glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) were measured, as well as the level of Nitric Oxide (NO), Nitric Oxide Synthase (NOS) and Endothelin 1 (ET-1) . The pathology of abdominal aortas was analyzed.Results Sixteen weeks after intervention, there was no significant difference between HIIT + folic acid group and the control group (P>0.05) . The levels of serum Hcy in the model group, folic acid group and the HIIT+folic acid group were (23.95±3.35) μmol/L,(8.73±0.60) μmol/L, and (6.19±0.34) μmol/L respectively (P<0.05) . Sixteen weeks after intervention, the content of MDA in HIIT+ folic acid group reduced, and there was no significant difference compared with the control group (P>0.05). The level of SOD and GSH-PX increased in HIIT+ folic acid group and folic acid group, and there was a significant difference compared with the model group. There were significant differences in activities of SOD and GSH-PX in HIIT+ folic acid group when compared with folic acid group (P<0.05). Compared with the control group, there were significant differences in levels of ET-1, NOS and NO in folic acid group (P<0.05), while there was no significant difference in the level of ET-1 and NOS between HIIT+folic acid group and control group (P>0.05) . Mild atherosclerotic lesions were observed in the HIIT+folic group. Conclusion High methionine diet can reduce the level of serum Hcy in HHcy rats, and high intensity interval exercise combined with folic acid intervention could reduce the level of serum Hcy, improve oxidative stress state, reduce the injury of endothelial function, and thus to alleviate atherosclerotic lesion.