A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18β-glycyrrhetinic acid(1), 3-oxo-15β-hydroxy-18β-glycyrrhetinic acid (2), 7β,15α-dihydroxy-18β-glycyrrhetinic acid (3), 3-oxo-7β, 15α-dihydroxy-18β-glycyrrhetinic acid (4), 7β-hydroxy-18β-glycyrrhetinic acid(5) and 15α-hydroxy-18β-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation
Biotransformation ; Cunninghamella ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; metabolism ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

AIM: To study the effect of Yupingfeng Powder(YPFP) on rat's allergic rhinitis induced by stimulating ovalbumin.and reveal the influence on Th1,Th2 and Th1/Th2 proportion. METHODS: 8-week-old BALB/c mice were sensitized by means of intranasal and systemic intraperitoneal injection application of OVA,6 subjects were administered including 2,500 mg/kg extracts of YPFP for 7 days in early stage(interference group);6 mice were administered YPFP in after challenge(therapy group);the placebo group were given saline.After 7 days,Th1 and Th2 in splenocyte were detected by flow cytometry(FCM) and nasobuccal mucosa pathology were observed. RESULTS: When compared with Th2 of animal model group(9.86?1.40),there was a significant decrease expressing Th2 after PFP treatment(3.41?0.72,P

AIM: To observe the efficacy and safety of intravitreal injection of Conbercept for the treatment of wet age-related macular degeneration( AMD) .
METHODS:Retrospective analysis. A total of 20 patients involving 22 eyes were diagnosed of wet AMD and confirmed by routine ophthalmic examination, fundus fluorescein angiography ( FFA ) and optical coherence tomography. All these affected eyes received intravitreal injection of 10 mg/ml of 0. 5mg Conbercept, once monthly, for 3 successive times during the initial treatment. The need for repeated treatment was determined according to patients'disease conditions. The patients were followed up once monthly for ≥6mo. The changes in best corrected visual acuity ( BCVA ) , central retinal thickness ( CRT ) and choroidal neovascularization ( CNV) lesion leakage of the affected eyes before and after treatment were compared and analyzed.
RESULTS:Within 1, 3 and 6mo after treatment, the mean BCVA ( logMAR ) of the affected eyes increased when compared with before treatment;the difference was statistically significant(P<0. 01). In 1, 3 and 6mo after treatment, the mean CRT of the affected eyes decreased when compared with before treatment;the difference was statistically significant(P<0. 01). During the last follow-up, FFA showed that macular CNV lesion leakage disappeared in 20 eyes(90%) while leakage mitigated in 2 eyes ( 9%) . During the follow - up, there were no treatment - related serious ocular complications and systemic serious adverse reactions.
CONCLUSION: Clinically, intravitreal injection of Conbercept for the treatment of wet AMD can increase visual acuity of the affected eyes. It also can decrease CRT of the affected eyes, and inhibit neovascular leakage. There are no treatment-related adverse reactions.

OBJECTIVETo study the changes of urine metabolites in hypertension patients of ascendant hyperactivity of Gan yang syndrome (AHGYS), and to explore its essence in hypertension patients.

METHODSTen typical hypertension patients of AHGYS were recruited as the patient group, and the other twelve healthy volunteers were recruited as the normal group. The metabolite profiling in the urine were collected using by high performance liquid chromatography coupled with time of flight mass spectrometry (HPLC-TOFMS). The principal component analysis (PCA) and partial least-square discriminant analysis (PLS-DA) were analyzed using SIMCA-P Software. The differential metabolites in the urine were found out and identified. The possible relevant metabolic pathways were explained.

RESULTSThe data from the analysis by PCA in the urine samples of the patient group and the normal group showed, two sets of data could be obviously classified in the score plot. Compared with the normal group, significant changes happened to the body metabolism in the patient group. The metabolites relevant to hypertension patients of AHGYS were determined using the PLS-DA. Fifteen compounds of the structure and metabolic pathways had been confirmed through inquiring KEGG Database, mainly including amino acids, free fatty acids, sphingosine, and so on.

CONCLUSIONSThe hypertension patients of AHGYS were studied using HPLC-TOFMS combined with pattern recognition, thus finding out small molecular metabolic markers from the microscopic field, which was advantageous in probing the biological nature of Chinese medicine syndromes.

Adult ; Aged ; Case-Control Studies ; Chromatography, High Pressure Liquid ; methods ; Discriminant Analysis ; Female ; Humans ; Hypertension ; diagnosis ; urine ; Least-Squares Analysis ; Male ; Mass Spectrometry ; methods ; Medicine, Chinese Traditional ; methods ; Metabolome ; Middle Aged ; Principal Component Analysis

OBJECTIVETo study the analgesic mechanism of pricking blood therapy at Ashi points in the acute gouty arthritis rat.

METHODSForty rats were randomly divided into a blank group, a model group, an indomethacin group and a pricking blood group. Except the blank group, other groups were injected with sodium urate liquor into the ankle cavity to develop the acute gouty arthritis rat model, and the indomethacin group received gastric perfusion of indomethacin, and the pricking blood group were treated with pricking blood therapy at Ashi points. The peripheral pain mediums K+, NE, DA, (5-HT contents were determined.

RESULTSThe K+, DA, 5-HT contents in the pricking blood group decreased significantly as compared with the model group (P < 0.01, P < 0.05); there was no significant difference in the content of NE between the pricking blood group and the model group.

CONCLUSIONPricking blood at Ashi points can effectively inhibit release of the peripheral pain mediums K+, DA and 5-HT.

Animals ; Arthritis, Gouty ; therapy ; Pain ; Rats ; Stomach

OBJECTIVE:To investigate the differences of the active components of different parts of rhubarb(the head,the body and the end part of the root)from genuine medicinal materials in Gansu province so as to provide theoretical basis for its quality control.METHODS:A comparative study of the fingerprints of different parts of rhubarb extract were conducted by HPLC,in which,the C18 was used as the chromatographic column under room temperature,the mobile phase consisted of menthol-0.1%H3PO4(70∶30)with a flow rate of 1.0ml/min,the column's temperature of room temperature and detection wavel_ ength of 280nm.RESULTS:Significant differences were noted in the HPLC fingerprints of different parts of rhubarb,there were significant differences in the contents of the active components such as emodin,chrysophanol,etc.,the contents decreased progressively from the head of root,the body part to the end part.CONCLUSION:In order to ensure the medical value and the quality of rhubarb medicinal material,different parts of which should be differentiated in the quality control.

OBJECTIVETo observe clinical effects of posterior short-segment fixation with undermining decompress by posterior ligament complex for the treatment of upper lumbar burst fractures.

METHODSFrom October 2010 to March 2013,23 patients with upper lumbar burst fractures (Denis B type) were treated by posterior short-segment fixation with undermining decompress by posterior ligament complex. There were 18 males and 5 females aged from 26 to 64 years old with an average of 45.7 years old. Twelve patients were caused by falling down, 5 cases were caused by traffic accident, 4 cases were the bruise injury caused by heavy object and 2 cases were caused by other injury. Fourteen patients were L1 fracture and 9 patients were L2 fracture. Thirteen patients were combined with nerve injuries (degree D according to ASIA classification). Internal fixation were removed from 12 to 20 months with an average of 14.3 months. JOA scores and imaging changes were recorded and compared at different time points.

RESULTSAll patients were followed up from 18 to 24 months with an average of 20.4 months. Thirteen patients with nerve injuries were completely recovered at 3 to 6 months after operation. JOA score at 1 year after operation was 20.63 ± 0.92, and 20.38 ± 1.06 at 3 months after removal of internal fixation,which were improved obviously than 9.90 ± 2.73 at 3 months after operation. (P > 0.05) Anterior height of injured vertebrae, vertebral body angle and local Cobb angle was (95.0 ± 0.53)%, (2.78 ± 1.36) and (2.43 ± 1.52) °respectively, and improved obviously than that of before operation (P < 0.05). There was no statistical significance in JOA scores at 3 months after removal of internal fixation and 1 year after operation (P > 0.05).

CONCLUSIONposterior short-segment fixation with undermining decompress by posterior ligament complex for the treatment of upper lumbar burst fractures has advantages of minimally invasive, could effective recover vertebrae height, maintain stability of spine, decrease low back pain. It is a safe and effective operative method.

Adult ; Decompression, Surgical ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Spinal Fractures ; surgery

OBJECTIVETo explore the effect of IL-28B variation on the response of patients with chronic hepatitis C virus (HCV) infection to therapy.

METHODSA total of 220 patients with chronic hepatitis C (CHC) were prospectively treated with pegilated interferon (peg-IFN) in combination with ribavirin (RBV) for 48 weeks. After completing the therapy, the patients were followed-up for 24 weeks and the therapeutic effectiveness was evaluated. The rs8099917 was identified from each cohort. The IL28B genotype was compared in hepatitis C patients to assess the effect of single nucleotide polymorphism (SNP) on different treatment response.

RESULTThe proportion of the rs8099917 TT, TG, and GG genotypes was 71.4%, 25.0%, and 3.6% in sustained viral response (SVR) group; 15.8%, 60.5%, 23.7% in null response (NR) group; 38.1%, 52.3%, 9.6% in relapse (RP) group. There was a statistically significant difference in the genotype among SVR, NR and RP groups (P < 0.001, Chi-square test). NR vs. SVR (TG vs. TT: OR = 7.67, 95% CI: 2.91-20.56, P < 0.001). RP vs. SVR (TG vs. TT: OR = 3.10, 95% CI: 1.14-6.36, P < 0.01).

CONCLUSIONSThe genotypes of IL-28 B (rs8099917) is closely related to the effectiveness of peg-IFN-alpha/RBV therapy, and it is an important predictive factor before treatment in patients with chronic hepatitis C.

Adult ; Aged ; Antiviral Agents ; therapeutic use ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Genetic Variation ; Hepacivirus ; drug effects ; physiology ; Hepatitis C, Chronic ; drug therapy ; genetics ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Interleukins ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Ribavirin ; therapeutic use ; Treatment Outcome

Objective To measure the concentration of intracellular free Ca2+ ([Ca2+]i) in neuron-like cells resulted from rat bone mesenchymal stem cell (BMSCs) differentiation induced by salvia miltiorrhiza injection and provide some theoretical basis for the BMSCs transplantation. Methods The rat BMSCs were separated from rat bone marrow and cultured in vitro. After induced by basic fibroblast growth factor and 10mL/L salvia miltiorrhiza injection, the cells were identified with immunofluorescence staining against NeuN. The same procedure was performed on primarily cultured hippocampal neurons. Then, the [Ca2+]i of the differentiated neuron-like cells was determined and compared with primarily cultured hippocampal neurons. Results The BMSCs after induced by basic fibroblast growth factor and salvia miltiorrhiza injection expressed neuronal phenotypes similar to the cell appearance of neurons with NeuN. The average fluorescence intensity of the neuron-like cells derived from BMSCs was 984.75±79.51, while the average fluorescence intensity of the primarily cultured hippocampal neurons was 769.42±60.93. No significant difference was found between them (P>0.05). Conclusion The neuron-like cells from rat BMSCs differentiation induced by salvia miltiorrhiza injection possess certain neuronal properties.

Objective To explore the effects of salvia miltiorrhizae (SM) injection on the apoptosis of cultured rat neuronal stem cells induced by hypoxia and the activity of Caspase-3, in order to provide the further evidence for the molecular mechanism of neuroprotection of SM injection. Methods The neuronal stem cells from neonatal rat hippocampus were cultured and divided randomly into normal control group, hypoxia group and SM treatment group. After Hoechst staining, the apoptotic morphological change and apoptosis percentage were observed under fluorescence microscope. The activities of Caspase-3 in the 3 groups were evaluated by the colorimetric assay. Results Compared with normal control group [(2.75±0.28)%, 1.16±0.07], the percentage of apoptosis and the activity of Caspase-3 were increased significantly in neuronal stem cells cultured in hypoxia [(30.12%±2.09)%,3.85±0.41, P<0.05). Application of SM injection reduced markedly the percentage of apoptosis and the activity of Caspase-3 of the neuronal stem cells cultured in hypoxia [(9.16±1.34)%, 1.50±0.09, P<0.05].Conclusion SM injection can depress the apoptosis of the rat neuronal stem cells induced by hypoxia,so as to exert the neuroprotection.