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 explore the efficacy and safety of greenlight photoselective vaporization of prostate(PVP) in high-risk older patients with benign prostatic hyperplasia(BPH).Methods A total of 96 high-risk older patients with BPH underwent PVP.The operative time,blood loss,indwelling catheterization and operative complications were observed.The variables such as IPSS,QOL,uroflowmetry,post-void residual urine volume(RUV) were recorded and calculated pre-and post-operatively.Results All the 96 patients safely got through perioperative period.The mean operative time was(24.8?6.5)min,intraoperative blood loss was(55.4?16.8)ml and postoperative indwelling catheterization was(1.6?0.7)d.IPSS and QOL scores decreased form preoperative 29.8?5.2 and 5.2?0.8 to postoperative 8.4?2.3 and(1.4?)0.5,respectively.Qmax increased from(5.6?2.8)ml/s to(16.7?3.2)ml/s,and RUV decreased from(168.0?22.5)ml to(24.6?5.81)ml.There were significant differences of these parameters between pre-and post-operation(P

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.

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 observe the dynamic serological test results of 28 patients with acute brucellosis,and to investigate the relationship between serological test results and diagnosis,curative effect and prognosis of brucellosis.Methods Twenty eight patients(infected with sheep brucellosis) with acute brucellosis in the Department of Brucellosis in Heilongjiang Provincial Land Reclamation Headquarters General Hospital were selected as research subjects,and their serological changes were tested by means of tube agglutination test (SAT)and clinical outcomes were compared before and after each stage of treatment.In addition,symptoms of fever,weakness,sweating,joint pain,swollen lymph nodes and biochemical parameters [alanine aminotransferase (ALT),lactate dehydrogenase (LDH)] were also tested 3,6 and 9 weeks after the treatment.Results Antibody titer reached the peak at the third week,1 ∶ 400(++),which accounted for 39.29％ (11/28); 2857％(8/28) of the patients became negative at the sixth week; 50.00％ (14/28) became negative at the ninth week.Before the treatment,20(71.43％) patients had the symptom of fever,8 (28.57％) patients had the symptom of hyperhidrosis,28 (100.00％) patients had the symptom of joint pain,7(25.00％) patients had the symptom of lymph node enlargement,28 (100.00％) patients' ALT was elevated,and 26(92.86％) patients' LDH was elevated.After three weeks of treatment,except the three patients (10.71％) who occasionally had fever,the rest of the patient's temperature was returned to normal.Also the numbers of patients with the symptoms of fatigue,sweating and joint pain were significantly reduced,and specifically,the conesponding number was 13(46.43％),2(7.14％),and 21 (75.00％)patients,respectively.ALT and LDH returned to normal(only one patient's ALT was out of the range).At the sixth week,all the patients' symptoms of fever and hyperhidrosis disappeared.The number of patients with the symptoms of joint pain and lymph node enlargement reduced to 12(42.86％) and 3(10.71％),respectively.The results of biochemical tests(ALT and LDT) returned to normal.At the ninth week,most patients' clinical symptoms disappeared.A few patients still had the symptoms of weakness[2(7.14％)] and joint piin[6(21.43％)].Conclusions After effective treatment,antibody titer of patients decreases rapidly,at the same time,the clinical symptoms improve quickly.There is a parallel relationship between the change of antibody titer and clinical symptoms.It is demonstrated that the appearing time of brucellosis specific antibodies,the ampfitude and speed of change of antibody titers can be used in diagnosis,therapeutic evaluation and prognosis of the disease.

Objective To observe the clinical features of brucellosis spondylitis and analyse the reasons for its misdiagnosis,and improve the level of diagnosis and differential diagnosis.Methods Forty-two clinically diagnosed patients with brucellosis spondylitis were studied retrospectively,and these patients were diagnosed and hospitalized in the General Hospital of Heilongjiang Land Reclamation Bureau.Their medical records were analyzed,which included the general information,medical history,clinical symptoms,results of magnetic resonance imaging(MRI) and serum tube agglutination test(SAT) and so on.Results Main clinical symptoms and signs were severe persistent neck,back and leg pain.They also had plate shape low back but without kyphosis.In addition,patients had to keep in one posture because their spinal activity was limited.Also,scoliosis or pelvic tilt and lameness may occur when standing,which were typical symptoms of nerve root compression.Thirteen cases were diagnosed as tuberculosis,accounting for 30.95％(13/42); 6 cases were diagnosed as lumbar disc herniation,accounting for 14.28％ (6/42); 2 cases were diagnosed as ankylosing spondylitis,accounting for 4.76％ (2/42).Therefore,the total rate of misdiagnosis was 50％ (21/42).Abnormal MRI signal intensity can be seen in the pathological vertebrae.Specifically,T1-weighted images (T1WI) showed low signal,T2-weighted images (T2WI) showed high signal,or mixed high and low signal intensity was observed.Vertebral showed wedge deformation without collapse and sequestrum; strip and sheet abnormal signal can also be found within the intervertebral disc.Normal structure disappeared and disc space became narrow.Accordingly,the plane dural sac was compressed to form visible abscess near the spine,but psoas abscess was not found.Patients with positive SAT result accounted for 92.85％ (39/42).Conclusion Reasons for misdiagnosis include lack of detailed medical records,atypical clinical symptoms and similar imaging changes and so on.

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.

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