OBJECTIVETo evaluate the protective effects of punicosides on alcohol induced acute liver injury in mice and its possible mechanisms as well.

METHODThe 60 mice were randomly divided into normal control, model group, three dose groups of punicosides with low, medium and high, then there is silibinin group. Three dose groups of punicosides and silibinin were given in advance by gavage for 4 weeks, then the mouse model of alcoholic acute liver injury was established. The serum levels of ALT, AST and TG were determined, and the mice were killed to calculate somatic index of liver, thymus as well as spleen. MDA, SOD, GSH-Px and GSH-ST were detected in the liver homogenate. Histopathological changes of the liver were observed by HE staining. The expression of MCP-1 and NF-kappaB in the liver tissues were detected by immunohistochemistry.

RESULTMid and high dose of punicosides reduced the liver index in mice significantly, improved liver steatosis, decreased the level of ALT, AST and TG in serum and the content of MDA in liver homogenate, furthermore the two dose groups increased the activity of SOD, GSH-Px and GSH-ST, inhibited the expression of MCP-1 and NF-kappaB in liver tissue.

CONCLUSIONPunicosides can protect the acute liver damage induced by alcohol.

Alcohols ; adverse effects ; Animals ; Chemical and Drug Induced Liver Injury ; blood ; drug therapy ; metabolism ; pathology ; Chemokine CCL2 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation ; drug effects ; Liver ; drug effects ; enzymology ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; NF-kappa B ; metabolism

Abstract: Molecular genetic map is a fundamental organizational tool for genomic research. However, a genetic linkage map for Bupleurum chinense DC. has not been developed. In this study, with the theory of pseudo-testcross, 96 F1 plants from an intraspecific cross of B. chinense were used as mapping populations. Twenty eight ISSR (inter-simple sequence repeat) primers and 44 SSR (simple sequence repeat) primers were used to detect the polymorphisms between the parental plants, and of them, 28 ISSRs and 14 SSRs were selected to analyze the F1 populations. The map consisted of 13 linkage groups which included 80 (72 ISSRs and 8 SSRs) loci, and covered 2 633.9 cM with an average density of 33.4 cM. All 13 linkage groups consisted of 2-31 loci ranging in length from 15.4-1295.7 cM. This map will provide a basis for studies on gene mapping, map-based cloning and maker-assisted selection of important traits in B. chinense.

Objective To evaluate a combination of open surgery and multiple interventional methods in the treatment of DVT of the lower extremities. Methods 521 cases (521 limbs,356 male and 165 female) were studied in this group. Age ranged from 16 to 86 years with the mean age of (46 ±9)years. All 521 cases with DVT were treated by Fogarty embolectomy catheter. Among them,348 cases underwent percutaneous transluminal angioplasty (PTA), 135 cases received PTA and ultrasound ablation,stent-grafts were implanted in 108 cases. Results Based on angiography during operation, the obstructed iliofemoral vein received complete recanalization in 511 cases. Among them, the postoperative luminal diameter was more than 90% in 38 cases after Fogarty embolectomy, the average stenosis rate was reduced from 90% ±5% to 24% ±5% in 365 cases after PTA and stent-grafts were implanted in 108 cases with the stenosis rate still over 50% after PTA. Only partial recanalization was achieved in the entrance of common iliac vein to inferior vena cava in 10 cases. Of the 521 cases,472 cases were followed-up with mean time of (53 ± 26) months, ranging from 8 to 108 months and 462 cases reported satisfactory results with normal life,the unsuccessful 10 cases still felt swelling pain especially in erect position. Complications occurred in 33 cases. Conclusions Open surgery combining with multiple interventional methods is a safe and effective method in the treatment of DVT.

Objective:To establish the fingerprint analysis method for the aqueous extracts of Eucommia ulmoides from Enshi by HPLC. Methods:The fingerprint of aqueous extracts of ten batches of Eucommia ulmoides from Enshi were analyzed by HPLC. The columnwasWondaSilC18(250mm×4.6mm,5μm). Themobilephaseconsistedofacetonitrile-0.1% phosphoricacidwithgradient elution. The flow rate was 1. 0 ml·min-1 , the detection wavelength was 230 nm, the column temperature was 25℃, and the injection volume was 10 μl. Results:The fingerprint consisted of 12 common peaks. The similarity range of ten batches of Eucommia ulmoides calculated by similarity evaluation system for the chromatographic fingerprint of TCM(Version 2004 A)was 0. 596-0. 997. The standard fingerprint of Eucommia ulmoides was established by HPLC. Conclusion: The established HPLC fingerprint analysis method for Eu-commia ulmoides from Enshi is simple, stable and reproducible, which can effectively control the quality of Eucommia ulmoides from Enshi.

Objective To investigate antimicrobial resistance and ceftriaxone resistance mechanisms in clinically isolated nontyphoidal Salmonella(NTS),and provide evidence for the prevention and control of NTS infection and rational use of antimicrobial agents.Methods 108 NTS isolates were isolated from stool specimens of outpatients with acute diarrhea in the Second Hospital of Tianjin Medical University and Tianjin Medical University General Hospital from May to October of 2014,NTS were performed antimicrobial susceptibility testing;non-ceftriaxone-susceptible isolates were typed by serological,multilocus sequence (MLST),and pulsed-field gel electrophoresis (PFGE)methods,extended-spectrumβ-lactamase (ESBL)detection and AmpC genes were detection.Results Among 108 NTS isolates,mono-drug resistance rate to 11 antimicrobial agents was 49.07% (n= 53),multidrug resistance rate was17.59% . Susceptibility rates to nalidixic acid,levofloxacin,ciprofloxacin,ceftriaxone,and ertapenem were 61.11% ,66.67% ,68.52% ,97.22% ,and 100.00% respectively. Three non-ceftriaxone-susceptible NTS isolates were detected,2 were ST11 Salmonellaenterica serotype (Sa8709,Sa8771),1 was ST34 Salmonellatyphimurium serotype(Sa8763). Cluster analysis of PFGE revealed that Sa8709 was highly similar to Sa8771 strains(91 .70% ), but the similarity to Sa8763 was low(55.80% );Sa8709 strain carried CTX-M gene,Sa8771 strain carried CTX-M and TEM genes,Sa8763 strain carried OXA gene. Conclusion Clinically isolated NTS in this area are low resistant to fluoroquinolones,multidrug resistant strains carrying ESBLs have emerged.

Objective To study the effect of neuronal Nogo-66 receptor (NgR1) antagonist,soluble Nogo-66 receptor (sNgR1-Fc),on promoting the endogenous neural precursor cells (NPCs) differentiating into neurons in order to clarify the mechanism.Methods The cortical infarction was induced by photochemistry,named photothrombotic cortical injury (PCI).Twelve Sprague Dawley rats were randomly divided (random number) into three groups:Sham-operated group,PBS group,and sNgR1-Fc group.PBS (PBS group) or sNgR1-Fc (sNgR1-Fc group) was injected into the lateral ventricle of brain with a minipump.BrdU (Bromodeoxyuridine) was injected into the peritoneal cavity 4-6 days after PCI.The subdentate gyrus zone (SGZ) of brain from sacrificed rat was harvested for Immunohistochemistry to observe the ratio of NeuN +/BrdU + cells 35 days after PCI.Proteins including Nestin、Notch1 and Mash1 were detected by Western Blot.Results The cortical infarction in rat was successfully induced by photochemistry.Thirty-five days after PCI,the BrdU + cells number and theratio of NeuN +/BrdU + in the SGZ of the ipsilateral cerebrum hemisphere with PCI were significantly higher in sNgR1-Fc group than those in PBS group (P ＜ 0.05).The levels of Notch1,Mash1 and Neuro D in the sNgR1-Fc group were significantly higher than those in the PBS group (P ＜ 0.05),which were significantly higher than those in the Sham-operated group.Conclusions sNgR1-Fc could promote the endogenous NPCs differentiating into neurons in a cortical infarction model.The mechanisms may be attributed to the Notch/bHLH (proneural basic helix-loop-helix genes) signaling way.

This article firstly established a new efficient method for screening anti-osteoporosis ingredients, which used two-dimensional zebrafish model combined with hyphenated chromatographic techniques to evaluate anti-osteoporosis activities of epimedin A and its metabolite baohuoside I. Adult zebrafish was used for metabolism of epimedin A in 0.5% DMSO, and LC-MS was used for analysis of the metabolite, which was captured by HPLC, and prednisolone-induced osteoporosis model of zebrafish was used to evaluate the anti-osteoporotic activities of trace amounts of epimedin A and baohuoside I. The results indicated that epimedin A and baohuoside I can prevent prednisolone-induced osteoporosis in zebrafish. The developed method in this paper enables the separation, enrichment and analysis of micro-amount metabolite of epimedin A, and anti-osteoporosis activities in vivo of epimedin A and baohuoside I was simple and efficient screening resorting to zebrafish osteoporosis mode. This paper would provide new ideas and methods for a rapid and early discovery of anti-osteoporosis activities of micro-ingredients and its metabolite of traditional Chinese medicine.
Animals ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; metabolism ; pharmacology ; Mass Spectrometry ; Osteoporosis ; drug therapy ; Zebrafish

OBJECTIVETo break through the restrictions of the evaluation model and the quantity of compounds by using the two-dimensional zebrafish model combined with chromatographic techniques, and establish a new method for the high-throughput screening of active anti-osteoporosis components.

METHODAccording to the research group-related studies and relevant foreign literatures, on the basis of the fact that the zebrafish osteoporosis model could efficiently evaluate the activity, the zebrafish metabolism model could efficiently enrich metabolites and the chromatographic techniques could efficiently separate and analyze components of traditional Chinese medicines, we proposed that the inherent combination of the three methods is expected to efficiently decode in vivo and in vitro efficacious anti-osteoporosis materials of traditional Chinese medicines.

RESULT AND CONCLUSIONThe method makes it simple and efficient in the enrichment, separation and analysis on components of traditional Chinese medicines, particularly micro-components and metabolites and the screening anti-osteoporosis activity, fully reflects that efficacious materials of traditional Chinese medicines contain original components and metabolites, with characteristic of "multi-components, multi-targets and integral effect", which provides new ideas and methods for the early and rapid discovery of active anti-osteoporosis components of traditional Chinese medicines.

Animals ; Chromatography ; methods ; Disease Models, Animal ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; trends ; Osteoporosis ; drug therapy ; physiopathology ; Phytotherapy ; methods ; trends ; Reproducibility of Results ; Zebrafish ; physiology

Objective To observe the protective effects of soluble Nogo-66 receptor (NgR1 )antagonist (sNgR1-Fc) on cortical axons after cortical infarction in rats,and to study the phenomenon and molecular mechanism of its protective effects on and regeneration of axons.Methods The cortical infarction was induced by photochemistry,termed photothrombotic cortical injury (PCI).Fifteen Sprague Dawley rats were randomly divided into three groups:Sham-operated group,PBS (phosphate buffered solution) group,and s-NgR1-Fc group.In PBS group,PBS was injected into the lateral ventricle of rats; and in sNgR1-Fc group,sNgR1-Fc was injected instead of PBS. The ipsilateral cortex with lesion was harvested for histomorphometry and transmission electron microscope observation 7 days after PCI. Proteins including GTP-RhoA,p-JNK,p-c-JUN and p-ATF-2 were detected by Western blot,as well as Total-J and Total-RhoA.Results The cortical infarction in rats was successfully induced by photochemistry.Compared with sham-operated group,the pathological changes in PBS groups were more serious,including extensive edema or disappearance of axoplasm of fiber without medulla sheath involved and extensive thickening or layer derangement in axoplasm of fiber with medulla sheath involved.These changes were improved significantly after sNgR1-Fc treatment.The levels of GTP-RhoA,p-JNK1,p-JNK2,p-c-JUN and p-ATF-2 in the PBS group were significantly higher than those in the sham-operation group ( P ＜ 0.05 ),whereas the levels of Total-RhoA,Total-JNKl and Total-JNK2 were not different significantly between these two groups (P ＞0.05 ).The sNgR1-Fc treatment up-regulated the levels of these proteins ( P ＜ 0.05 ).Conclusions There is pathological change in axon induced by cerebral hypoxia-ischemia for a long period after cortical infarction.The mechanisms may be associated with RhoA/ROCK/JNK/c-Jun signal way,which is activated by ischemia injury and related to the inhibition of regeneration in axon.Our study shows that NgR1-Fc may inhibit this pathway significantly,and then promote the regeneration of axon partially.

OBJECTIVE To investigate the bone development activity and differences in safety of ethanol extract (EE) and water decoction (WD) of Psoralea corylifolia L.efficiently.METHODS Zebrafish larvae were co-exposed to prednisolone 25 μmol· L-1 and different concentrations of EE and WD (0.1,1.0,10 and 100 mg crude drug· L-1),and etidronate disodium (ED) 30 mg·L-1.All these groups were incubated at 28.5℃ until 9 dpf.The medium solution was changed every other day.Zebrafish skeleton at 9 dpf was stained with alizarin red and inspected under an optical microscope,in addition,the death toll and organ toxicity of zebrafish were also observed.The mRNA expression of osteoprotegrin (OPG) and receptor activator of NF-κB ligand (RANKL) in 9 dpf zebrafish were determined with fluorescence quantitative PCR.Zebrafish embryos (1 dpf) were exposed to various concentrations of EE (10,20,30,35,40,50 and 60 mg crude drug· L-1),WD (10,50,100,125,150,175,200 and 500 mg crude drug· L-1),psoralen (12.5,25.0,50.0,100.0,200.0 and 400.0 μmol·L-1) and bakuchiol (1,5,10,25 and 50 μmol· L-1).Embryonic morphology of zebrafish (3 dpf) was inspected with an optical microscope and the death toll of embryos or larvale was counted from 2 dpf to 9 dpf and LC50was calculated.Components of EE and WD ware analyzed by HPLC method.RESULTS Both EE (0.1 mg crude drug· L-1) and WD (1.0 mg crude drug· L-1) groups could increase the staining area and optical density values of zebrafish skeleton compared with prednisolone group (P＜0.01),indicating the increase in bone mineralization;the OPG mRNA expression in both EE and WD (1.0 mg crude drug· L-1) groups increased,while the RANKL mRNA expression decreased (P＜0.01) and the ratio of OPG/RANKL improved obviously (P＜0.01).Embryos exposed to EE,WD,psoralen and bakuchiol showed swelling of the heart and yalk sac,and decrease in GOT.The LC50 of WD and psoralen was 5～8 and 5～21 times that EE and bakuchiol,respectively.The composition and relative content of EE and WD also varied considerably.CONCLUSION Bone development activity and toxicity of EE are both stronger than those of WD.The lipid soluble characteristic components of Psoralea corylifolia L.,may be critical components of toxicity.