Objective To investigate the effect of tourniquet compression on axonal transport in sciatic nerve of rats.Methods Twenty-four 12-week old male SD rats weighing 250-300 g were randomly divided into 4groups according to the duration of tourniquet compression(n=6 each):1,2,4 and 12 h.The tourniquet was applied to the middle 1/3 of thigh.In each animal whether the left or right thigh was compressed was determined by a flip of coin.The tourniquet was released for 10 min after every hour of compression.A 3-cm segment of sciatic nerve was removed at the end of tourniquet compression(1.5 cm proximal and 1.5 cm distal to the site of compression).Immuno-histochemistry was used to measure the expression of insulin-like growth factor-1(IGF-1)in the sciatic nerve.The ratio of average optic density of the compressed sciatic nerve to that of control was used to estimate the degree of IGF-1 accumulation.The regression equation of the interaction between the duration of compression and accumulation of IGF-1 was analyzed.Results There was significant accumulation of IGF-1 in the sciatic nerve proximal to the compressed site.The accumulation increased with the duration of compression.There was no significant accumulation of IGF-1 in the sciatic nerve distal to the compressed site.The regression equation of the interaction between the duration of compression(X)and accumulation of IGF-1(Y)was Y=0.422X+0.887.Conclusion Tourniquet compression of sciatic nerve can inhibit axonal transport.The accumulation increases with the duration of compression.

To study the in situ intestinal absorption of five oligosaccharides contained in Morinda officinalis How. (sucrose, kestose, nystose, 1F-Fructofuranosyinystose and Bajijiasu). The absorption of the five oligosaccharides in small intestine (duodenum, jejunum and ileum) and colon of rats and their contents were investigated by using in situ single-pass perfusion model and HPLC-ELSD. The effects of drug concentration, pH in perfusate and P-glycoprotein inhibitor on the intestinal absorption were investigated to define the intestinal absorption mechanism of the five oligosaccharides in rats. According to the results, all of the five oligosaccharides were absorbed in the whole intestine, and their absorption rates were affected by the pH of the perfusion solution, drug concentration and intestinal segments. Verapamil Hydrochloride could significantly increase the absorptive amount of sucrose and Bajijiasu, suggesting sucrose and Bajijiasu are P-gp's substrate. The five oligosaccharides are absorbed mainly through passive diffusion in the intestinal segments, without saturated absorption. They are absorbed well in all intestines and mainly in duodenum and jejunum.
Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Intestinal Absorption ; Intestine, Small ; metabolism ; Male ; Morinda ; chemistry ; Oligosaccharides ; chemistry ; pharmacokinetics ; Perfusion ; Rats ; Rats, Sprague-Dawley

Objective To analyze clinical diagnosis and management of 5 patients with actinomycete keratitis.Design Retro- spective case series.Participants 5 patients (5 eyes) with actinomycete keratitis.Methods The clinical features and microbiologic da- ta of 5 culture-proven cases of actinomycete keratitis recorded between October 2004 to March 2006 were analyzed.Main Outcome Measures clinical characteristics,isolations identification,drug susceptibility test and treatments.Results All patients were males and farmers.Of the 5 cases presented in this study,4 cases were followed by minor trauma as a predominant risk factor,and were pre- sented by a chronic progressive corneal ulcer with a wreath pattern of infiltrate.The diagnosis of all cases was based on laboratory in- vestigations,by which 4 cases of nocardia and one case of streptomyce were identified.A variable drug sensitivities were presented in nocardia isolates,which including TMP-SMZ,amicasin,gentamicin and fluorine-quinolones.Conclusions Nocardia keratitis is mainly followed by a minor trauma.It is identified predominantly by laboratory investigations.Tropical and systemically sensitive biotic are the initial choice,while debridement and amnionic transplantation could be an effective alternative.

OBJECTIVEThe selective loss of dopaminergic neurons in Parkinson's disease is suspected to correlate with the increase of cellular iron, which may be involved in the pathogenesis of PD by promotion of oxidative stress. This research investigated dopamine-induced oxidative stress toxicity contributed by iron and the production of dopamine-derived neurotoxins in dopaminergic SH-SY5Y cells.

METHODSAfter the SH-SY5Y cells were pre-incubated with dopamine and Fe2+ for 24 h, the cell viability, hydroxyl radical, melondialdehyde, cell apoptosis, and catechol isoquinolines were measured by lactate dehydrogenase assay, salicylic acid trapping method, thiobarbuteric acid assay, Hoechst 33258 staining and HPLC-electrochemical detection (HPLC-ECD), respectively.

RESULTS(1) Optimal dopamine (150 micromol/L) and Fe2+ (40 or 80 micromol/L) significantly increased the concentrations of hydroxy radicals and melondialdehyde in SH-SY5Y cells. (2) Induction with dopamine alone or dopamine and Fe2+ (dopamine/Fe2+) caused cell apoptosis. (3) Compared with untreated cells, the catechol isoquinolines, salsolinol and N-methyl-salsolinol in dopamine/Fe2+-induced cells were detected in increasing amounts.

CONCLUSIONDue to dopamine/Fe2+-induced oxidative stress similar to the state in the parkinsonian substantia nigra neurons, dopamine and Fe2+ impaired SH-SY5Y cells could be used as the cell oxidative stress model of Parkinson's disease. The catechol isoquinolines detected in cells may be involved in the pathogenesis of Parkinson's disease as potential neurotoxins.

Apoptosis ; drug effects ; physiology ; Catechols ; metabolism ; Cell Line, Tumor ; Cell Survival ; drug effects ; physiology ; Dopamine ; toxicity ; Dose-Response Relationship, Drug ; Humans ; Hydroxyl Radical ; metabolism ; Iron ; metabolism ; Iron Metabolism Disorders ; complications ; metabolism ; physiopathology ; Isoquinolines ; metabolism ; Malondialdehyde ; metabolism ; Models, Biological ; Nerve Degeneration ; chemically induced ; metabolism ; physiopathology ; Neurons ; drug effects ; metabolism ; Neurotoxins ; toxicity ; Oxidative Stress ; drug effects ; Parkinson Disease ; etiology ; metabolism ; physiopathology ; Salsoline Alkaloids ; metabolism ; Up-Regulation ; drug effects ; physiology

OBJECTIVETo determine the expressions of HO-2 and eNOS in the corpus cavernosum of castrated rats and investigate the correlation of HO-2 and eNOS with erectile dysfunction.

METHODSForty SD rats of 10 weeks were equally divided into 3 castrated groups (4, 8 and 12 weeks respectively after removal of bilateral testes) and a sham operation control group. All the rats were detected for serum testosterone (T), intracavernosal pressure (ICP) and mean arterial blood pressure (MAP) 2, 8 and 12 weeks after the surgery, the content of HO-2 analyzed by Western blot and the expressions of eNOS and HO-2 determined by immunohistochemistry.

RESULTSTestosterone level and ICP/MAP ratio were markedly decreased after the castration (P < 0.05). The HO-2 expression was detected in the corpus cavernosum of all but the 12-week castrated rats, and the area under the curve was (341.50 +/- 99.70) in the 4-week castrated group, significantly less than in the 8-week (705.00 +/- 152. 74) and the control group (876 +/- 443. 36) (P < 0. 05), with no obvious difference between the latter two. eNOS was found mainly in the vascular endothelial cells of the corpus cavernosum, significantly less in the castrated (123.94 +/- 30. 23) than in the control group (421.21 +/- 125. 12).

CONCLUSIONAndrogens might mediate the penile erectile function through HO-2 and eNOS in the corpus cavernosum of rats.

Animals ; Blood Pressure ; Blotting, Western ; Erectile Dysfunction ; blood ; enzymology ; physiopathology ; Heme Oxygenase (Decyclizing) ; metabolism ; Immunohistochemistry ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Orchiectomy ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood

OBJECTIVETo investigate the expression of Rho-kinase and heme oxygenase and the interaction among NOS/NO, HO/CO and RhoA/Rho-kinase in the corpus cavernosum of spontaneous hypertensive rats (SHRs).

METHODSA series of electric stimuli were applied to the corpus cavernosum nerves of 7 SHRs, the changes of ICP/MAP observed continuously and the expressions of ROCK2, HO-2 and eNOS analyzed by Western blot and immunohistochemistry. Another 7 WKY rats were taken as controls.

RESULTSCompared with the controls, ICP/MAP did not rise obviously (P < 0.05), HO-2 dropped sharply (P = 0.006) and the expression of the ROCK2 protein was elevated markedly (P = 0.017) in the SHRs. Immunohistochemistry showed HO-2 to be distributed in the smooth muscle cells and nervous cells of the corpus cavernosum, and eNOS mainly in the vascular endothelial cells. The expressions of HO-2 and eNOS were obviously reduced in the SHRs.

CONCLUSIONNOS/NO, HO/CO and RhoA/Rho-kinase are related with erectile dysfunction in SHRs and may interact on one another.

Animals ; Blotting, Western ; Erectile Dysfunction ; enzymology ; physiopathology ; Heme Oxygenase (Decyclizing) ; metabolism ; Hypertension ; enzymology ; physiopathology ; Immunohistochemistry ; Male ; Penis ; enzymology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; rho-Associated Kinases ; metabolism

SUMMARY Thehumanembryonicstemcells(hESCs)serveasaself-renewable,genetically-healthy, pluripotent and single source of all body cells,tissues and organs.Therefore,it is considered as the good standard for all human stem cells by US,Europe and international authorities.In this study,the standard and healthy human mesenchymal progenitors,ligament tissues,cardiomyocytes,keratinocytes,primary neurons,fibroblasts,and salivary serous cells were differentiated from hESCs.The human cellular health-safety of NaF,retinoic acid,5-fluorouracil,dexamethasone,penicillin G,adriamycin,lead ace-tate PbAc,bisphenol A-biglycidyl methacrylate (Bis-GMA)were evaluated selectively on the standar-dized platforms of hESCs,hESCs-derived cardiomyocytes,keratinocytes,primary neurons,and fibro-blasts.The evaluations were compared with those on the currently most adopted cellular platforms.Parti-cularly,the sensitivity difference of PM2.5 toxicity on standardized and healthy hESCs derived fibroblasts, currently adopted immortalized human bronchial epithelial cells Beas-2B and human umbilical vein endo-thelial cells (HUVECs)were evaluated.The results showed that the standardized hESCs cellular plat-forms provided more sensitivity and accuracy for human cellular health-safety evaluation.

Aim To determine the effect of exosomes from lipopolysaccharide-treated human bone marrow mesenchymal stem cells on proportion of Ly6Chigh and Ly6Clow monocytes/macrophages in inflammatory micro- environment. Methods BMSCs were obtained by gra-dient centrifugation, identified and then treated with li-popolysaccharide for 48 h. The exosomes were purified from conditional medium with or without LPS treatment and identified by CD63 protein using Western blot and transmission electron microscope. The diameters and concentration were detected by Nanoparticle Trafficking Analysis ( NTA ) . The monocytes/macrophages were sorted from bone marrow of the mice by magnetic beads. Cells were co-cultured with exosomes for 24 hours, and then treated with LPS for 48 hours. The proportion of Ly6C monocytes/macrophages was detec-ted by flow cytometry. Inflammatory cytokines in cell supernatant were investigated using ELISA. Results BMSCs surface markers CD44, CD90 were positively detected, but CD34, CD45 were not expressed. BM-SCs presented adipogenic differentiation ability. Exo-somes were positively expressing CD63 protein, and NTA showed that the diameters of exosomes were up to (82.4 ± 3.7 ) nm. BMSCs stimulated by LPS pro- duced more exosomes ( P < 0.01 ) . Exosomes from BMSCs with or without LPS treatment could increase the ratio of Ly6Chigh monocytes (P<0.01) and down-regulate the ratio of Ly6Chigh macrophages (P<0.05), and the effect of LPS treated-exosomes was more signif-icant than untreated-exosomes (P<0.05). Moreover, the concentration of IL-6 was also elevated under exo-somes treatment ( P <0.05 ) . Conclusions Human bone marrow mesenchymal stem cells-derived exosomes contribute to the regulation of Ly6Chigh monocytes/mac-rophages, indicating that they could be involved in the therapeutic treatment of inflammatory diseases.

OBJECTIVESynthesis and identification of complete antigen of rutin, the traditional Chinese medicine active ingredient, and develop rapid detection of rutin using enzyme-linked immunoassay method (ELISA). Immunogenicity of the complete antigen was also studied.

METHODPrepare the complete antigen by sodium periodate solution and identified by UV scanning and SDS-PAGE test. Male New Zealand white rabbits were immunized by the antigen to obtain the antiserum.

RESULTThe results of UV analysis showed that the coupling ratio of complete antigen is 13: 1. SDS-PAGE display of the artificial antigen was delayed compared with bovine serum protein. The titer of rutin antibody is 1:4 000. The sensitivity of IC50 was 5.37 mg x L(-1), the lowest detection limit was 1 mg x L(-1), the average recovery was 102%, the intra and interspecific RSD were less than 10%, cross-reactivity rate of antibodies and other analogs were less than 1%.

CONCLUSIONRutin complete antigen was synthesized successfully, and the rapid detection of rutin by ELISA method was successfully established.

Animals ; Antibody Specificity ; immunology ; Antigens ; immunology ; Cattle ; Cross Reactions ; immunology ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Immune Sera ; immunology ; Immunization ; Male ; Periodic Acid ; chemistry ; Rabbits ; Rutin ; chemical synthesis ; immunology ; Serum Albumin, Bovine ; immunology ; Solutions ; chemistry

OBJECTIVETo determine determinate five oligosaccharides, namely sucrose, 1-kestose, nystose, 1F-fructofurano-syinystose, bajijiasu contained in Morinda officinalis with an HILIC-ELSDI) method.

METHODWaters XBridge Amide (4.6 mm x 150 mm, 3.5 microm) hilic column was adopted for gradient elution, with acetonitrile (A) and 0.2% triethylamine (B) as the mobile phase. The column temperature was set at 40 degrees C, with the flow rate of 0.8 mL x min. Waters 2424 evaporative light scattering detector (ESLD) was used as detector, with the gas flow of 275.79 kPa and drift tube temperature of 90 degrees C.

RESULTThe detection range for the five oligosaccharides were 2.128-21.28 microg for sucrose (r = 0.999 3), 1.864-18.64 microg for 1-kestose (r = 0.999 6), 1.92-19.2 microg for nystose (r = 0.999 8), 1.912-19. 12 microg for 1F-fructofuranosyinystose (r = 0.999 5), 2.368-23.68 microg for bajijiasu (r = 0.999 4), respectively. The recovery of the five oligosaccharides ranged between 92.81%-102.8% (n = 6).

CONCLUSIONThe method is so simple, accurate and highly reproducible that it can be used as an analytical method for effective evaluation of the quality of M. officinalis herbs.

Chromatography, Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Morinda ; chemistry ; Oligosaccharides ; analysis ; Scattering, Radiation