Background Intravitreal injection with triamcinolone acetonide (TA) may cause complications,including increase of intraoculapressure (IOP),cataracand endophthalmitis.Ketorolatromethamine (Ketorolac) inew,lesadverse reactionof non-steroidal anti-inflammatory drug.The action mechanism of Ketorolaisimilato TA.Therefore,Ketorolamay be completely opartly replace Tin the treatmenof retinal edema.Objective The purpose of thistudy wato investigate the effectof Tand Ketorolaon the expressionof aquaporin-4 (AQP4) and vasculaendothelial growth facto(VEGF) in hypoxiretinal Müllecellin vitro and to explore the mechanism of treating retinal edemwith Tand Ketorolac.MethodThe propose of research and use of the animalwere approved by Animal ExperimenResearch Review Committee of Hubei University of Medicine.Twenty eyeof New Zealand albino rabbitwere extracted and the retinal tissue waisolated.The Müllecellwere cultured and passaged using the enzymatidigestion method and Müllecellwere identified using glial fibrillary acidiprotein (GFAP),vimentin and α-smooth muscle actin (α-SMA) by immunofluorescence staining.The hypoxicell modelwere established by culturing the cellin DMEM with 500 μmol/L CoCl2 fo0,6,12,24 hours.The cellof hypoxifo24 hourwere divided into normal control group,hypoxicontrol group,hypoxia+50,100,200 mg/L To50,100,200 mg/L Ketorolagroups.Corresponding drugwere added into the medium in the differengroups.The expressionof AQP4 mRNand VEGF mRNin Müllecellwere detected by semi-quantitative reverse transcription PC(RT-PCR).ResultThe cellgrew well and reached 80％ confluence with the irregulashape and ovoid nuclei 14-15 dayaftecultured.More than 95％ primary cellshowed positive reaction to GFAP,vimentin and α-SMA.The expressing levelof AQP4 mRNand VEGF mRNin Müllecell(values) were significantly differenin varioutime point(AQP4 mRNA:F=18.70,P＜0.01 ; VEGF mRNA:F =53.20,P＜0.01),and those of 6,12 and 24 houraftecultured with CoCl2were increased than those withouCoCl2 (P＜0.05).The expressing levelof AQP4 mRNand VEGF mRNin Müllecell(values) were significandifferenamong the normal control group,hypoxicontrol group,hypoxia+50,100,200 mg/L ToKetorolagroup(AQP4 mRNA:F =27.98,P ＜ 0.01 ; VEGF mRNA:F =10.03,P ＜0.01).Compared with the hypoxicontrol group,the expressing levelof AQP4 mRNand VEGF mRNin the Müllecellwere declined in the hypoxia+ 100,200 mg/L Tgroup and the hypoxia+100,200 mg/L Ketorolagroup (P＜0.05).The expressing levelof AQP4 mRNand VEGF mRNwere found statistically insignificandifference between hypoxia+ 100 mg/L Tgroup and hypoxia+ 100 mg/L Ketorolagroup,obetween hypoxia+ 200 mg/L Tgroup and hypoxi+200 mg/L Ketorolagroup (P＞ 0.05).ConclusionTand Ketorolacan downregulate the expressionof AQP4 and VEGF in Müllecellundehypoxiconditions,inferring thathey have similamechanism in the impacon AQP4 function in retinal edematoueye.

Background There are a lot of studies about the carrier of corneal endothelial transplantation,but the best carrier has not been defined.Objective This study was to investigate the biocompatibility of chitosan carrier with rabbit corneal endothelium in vivo.Methods Fresh eye-balls were obtained from 10 New Zealand white rabbits.Rabbit corneal endothelial cells (CECs) were isolated and cultured on chitosan carrier in vitro.The morphology and density of rabbits CECs were observed every day,and the expressions of fibronectin (FN),collagen-1 (Coil-I) and Zonula occludens 1 (ZO-1) were detected by immunoinfluorescence.The morphology and ultrastructure of CECs were observed under the scanning and transmission electron microscope.Chitosan carrier with CECs was implanted into the anterior chamber of the left eyes in ten healthy New Zealand white rabbits,and only paracentesis of anterior chamber was performed in the right eyes as controls.The inflammation of ocular anterior segment was examined under the slit lamp microscope,and corneal thickness was measured 1 week,4 and 8 weeks after operation.Corneal endothelium cell density and morphology were examined under the corneal endothelial microscope at postoperative 2 weeks.Corneal samples were collected for the regular histopathological examination to observe the inflammatory reaction at postoperative 1 month and 3 months.Paired t test was used for statistical analyses between the control group (left eyes) and the experimental group (right eyes).The use and care of the animals followed the Statement of ARVO.Results CECs formed an intact monolayer of cells with the uniform shape and size on the chitosan membrane after incubated for 5 days.The cells reached confluence of 90％ 7 days after cultured with the 40％ hexagon cells.Under the scanning electron nicroscope,rabbit CECs showed the round or polygon in the shape with the microvillus on the cell surface.The cells connected closely by desmosome.The processes,pseudopodiums and microvillus on the cellular surface,vacuole in the cytoplasm,expanded endoplasmic reticulum with ribosome and abundant chromatin were exhibited under the transmission electron microscope.The immunofluorescence examination revealed the positive expressions of FN,Coll-Ⅰ and ZO-1 in the CECs on the chitosan carrier.In the in vivo experiment,the exudation in the anterior chamber and corneal edema were seen under the slit lamp microscope 3 days after implantation of chitosan carrier with CECs.However,the inflammation was gone 14 days after operation.The differences of the corneal thickness were no significant between the experimental group and the control group 1 week and 4,8 weeks after operation (t =1.377,P=0.265;t =1.795,P=0.165 ; t =0.390,P =0.760).In addition,no significant differences were found in the CECs density and the hexagon cells rate between the two groups(P =0.365,0.062).The histopathological examination showed that the inflammatory cells around the chitosan membrane were disappeared 3 months after operation and showed a good corneal structure.Conclusions Chitosan carrier has a good biocompatibility with rabbit CECs and anterior chamber,and it may be a potentially good carrier for CECs transplantation.

OBJECTIVEIn order to understand the production mechanism of interferon and provide a scientific basis for preventionand clinical therapy, the expression changes of Toll-like receptor (TLR3) mRNA and the role of TLR3 in human lung epithelial cells (A549 cells) infected with respiratory syncytial virus (RSV) were investigated in this study.

METHODSRSV infected A549 cells were treated with or without specific antibodies of TLR3 and collected at the selected timepoints after RSV infection (4, 8, 12, 16 and 24h). The expressions of TLR3, IFN-alpha, IFN-beta and RSV F mRNA were evaluated by RT-PCR.

RESULTIt was found that RSV infection could markedly up-regulate the mRNA expression of TLR3, IFN-alpha, IFN-beta and RSV F protein in a time-dependent manner as the 24h mRNA expressions of them were 4 times, 3 times, 3 times and 0.7 times more than the basic expression, respectively. Treatment of TLR3 specific antibodies, whereas, significantly down-regulated the activation of TLR3. The mRNA expression of IFN-alpha and IFN-1beta also decreased accordingly and that of IFN-beta reduced more obviously than IFN-alpha, but that of RSV F protein rose significantly.

CONCLUSIONAbove data indicate that RSV infection could induce an apparent increase of antiviral genes of IFN-alpha and IFN-beta by activating TLR3 in human lung epithelial cells and the activated cells mediated Type I interferon is antiviral, which suggesting that TLR3 might play an important role in antiviral activity of RSV-infected human lung epithelial cells.

Epithelial Cells ; immunology ; virology ; Humans ; Interferon-alpha ; genetics ; Interferon-beta ; genetics ; Lung ; immunology ; virology ; RNA, Messenger ; analysis ; Respiratory Syncytial Viruses ; immunology ; Time Factors ; Toll-Like Receptor 3 ; genetics ; physiology ; Viral Fusion Proteins ; genetics

Objective To propose a new blood purification modality-hemodialysis with plasma- based dialysate (HD-PBD) plus high volume hemofiltration (HVHF) for patients with liver failure, and to evaluate the effect of this treatment on plasma cytokines.Methods Twelve patients with liver failure were included in this study.All patients received HD-PBD therapy in the first 6 hours,and then were treated with HVHF for 24 hours with the same filter (AV600).The levels of TNF-?,IL-1?, IL-6 and IL-8 in plasma before and after HD-PBD plus HVHF for 6 and 24 hours were examined respectively by ELISA,and changes of clinical parameters were observed at the same time point. Serum bilirubin,total bile acids (TBA),serum ammonia,blood urea nitrogen (BUN) and serum creatinine (Scr) were detected before and after treatment.Arterial blood gas analysis and the concentration of electrolytes were monitored before and after treatment.Results (1)HD-PBD for 6 hours was more effective than HVHF for 24 hours in removal of serum bilirubin and TBA(P＜0.05). (2)Serum ammonia,BUN,Ser,arterial blood HCO_3~-,PCO_2,PO_2 and electrolytes did not show significant difference before and after HD-PBD (P＞0.05),but these parameters significantly changed before and after HVHF (P＜0.05).(3)The average level of serum bilirubin was sharply decreased after HVHF for 24 h following HD-PBD(P＜0.05).(4)After HD-PBD plus HVHF,there was a marked reduction of the plasma levels of TNF-?,IL-6 and IL-8.Conclusions HD-PBD plus HVHF,a newly proposed modality for patients with liver failure,can effectively decrease serum bilirubin,TBA,BUN,Scr,ammonia and cytokines,and adjust water-electrolyte as well as acid- alkali balance.It is a low-cost,safe,simple and convenient therapy.

ObjectiveTo probe into the clinical efficiency of reconstruction to person who had composite tissue defect of the floor of mouth and the lower part of face with the free forearm flap and free iliac bone in the same term.MethodsBefore the surgery, the surgeon should carefully evaluate the scope and shape of defects in 11 cases who had the composite tissue defects of the floor of mouth and the lower part of face, to design the individual forearm flap, so that it matches with the defect region. During surgery, the first resumption of defects using titanium plates forming the basic shape of mandible and the occlusal relationship,then used to re-sawing to take modeling of the iliac bone,transplanted free iliac bone and fixed after the inside of the titanium plate, thereby restoring the continuity of mandible missing.ResultsFree forearm flap and free iliac bone all survived in 11 cases, the success rate of 100％. The patients were followed up for 6 to 12 months, although the shape of restoration areas were different levels of fat, but eating, swallowing and other oral function had been well improved, 7/11 could enter normal diet, 4/11 to enter liquid diet, At the same time, the patients in social activities can communicate in a language daily, sensory function of skin flap and facial appearance has been satisfactory recovery.ConclusionConformal free forearm flap and free iliac bone is an ideal way to reconstruct the composite tissue defects of the floor of mouth and the lower part of face in the same period, not only safe, practical prognostic effect, and also can significantly improve the patients quality of life, which is worthy of reference and use.

To investigate the relationship between the structures of methylhesperetin-7-alkyl ether analogues and their anti-inflammatory activities, nine new compounds, methyl-hesperetin (2), methylhesperetin-7-ethyl ether (3), 7-n-butyl ether (4), 7-n-hexyl ether (5), 7-n-octyl ether (6), 7-n-decyl ether (7), 7-n-dodecyl ether (8), 7-n-tetradecyl ether (9) and 7-n-hexadecyl ether (10), were synthesized with the lead compound of methylhesperidin (1). Their structures were confirmed by UV, 1H NMR, MS and HR-MS spectral data. The in vivo antiinflammatory activities of these compounds were tested on mouse paw edema induced by Freund's complete adjuvant (FCA) and mouse capillary permeability induced by acetic acid with po dose of 300 mg x kg(-1) x d(-1). The result indicated that the anti-inflammatory activities of the synthetic compounds increased firstly and then decreased with the elongating of the length of alkyl chain. After 25-day oral administration of compounds 6, 7 and 8, the inhibitory rates on mouse paw edema of adjuvant arthritis (AA) were 31.9%, 38.5%, 39.1%, respectively. They showed the concentrations of COX-2 in serum of AA mice respectively were 79.3, 75.4, 73.9 ng x L(-1) and the concentrations of PGE2 were in correspondence with 275.4, 258.9, 242.6 ng x L(-1). The inhibitory rates of compounds 6 and 7 on mouse capillary permeability induced by acetic acid were, respectively, 42.4% and 41.5% after 5-day oral administration. Compared with the lead compound of methylhesperidin, the anti-inflammatory activities of compounds 6, 7 and 8 were increased and showed an effective inhibition on the symptom of adjuvant arthritis and capillary permeability in mice.
Acetic Acid ; Animals ; Anti-Inflammatory Agents ; chemical synthesis ; chemistry ; pharmacology ; Arthritis, Experimental ; blood ; chemically induced ; drug therapy ; metabolism ; Capillary Permeability ; drug effects ; Cyclooxygenase 2 ; blood ; Dinoprostone ; metabolism ; Edema ; chemically induced ; drug therapy ; Female ; Freund's Adjuvant ; Hesperidin ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Male ; Mice ; Molecular Structure ; Random Allocation

OBJECTIVEImpulsivity is one of the core symptoms of children with attention deficit hyperactivity disorder (ADHD). In order to understand the neuromechanism of the impulsive behaviors in ADHD children, this study investigated the specific functional areas of the brain by functional MRI.

METHODSThe subjects consisted of 10 ADHD children with impulsivity, 7 ADHD children without impulsivity and 9 normal children. A functional MRI examination was performed when the subjects were instructed to finish GO and STOP tasks with the GO-STOP impulsivity paradigm. The MRI data during the two tasks of GO and STOP were averaged and the corresponding activation regions between groups were compared.

RESULTSThe data from the GO task revealed that the main activation regions of the normal children included frontal pole (superior frontal gyrus, middle frontal gyrus and medial frontal gyrus); the main activation regions of ADHD children without impulsivity were cerebellum (posterior lobe and anterior lobe bouton) and cingulated gyrus; those of ADHD children with impulsivity were medial globus pallidus and insula. The data from the STOP task showed that the main activation regions of normal children included superior frontal gyrus and middle frontal gyrus; those of ADHD children without impulsivity were middle frontal gyrus and cingulate gyrus; those of ADHD children with impulsivity were uncus and putamen. The activation regions of ADHD children with impulsivity were much fewer than the other two groups.

CONCLUSIONSThe behavior of impulsivity-control involves a number of specific functional areas in the cerebral cortex. Compared with normal children, ADHD children without impulsivity have weaker brain function and brain activation, and ADHD children with impulsivity demonstrate much fewer brain activation regions, worse brain function and little awareness of the cerebral cortex.

Adolescent ; Attention Deficit Disorder with Hyperactivity ; physiopathology ; Brain ; physiopathology ; Child ; Humans ; Impulsive Behavior ; physiopathology ; Magnetic Resonance Imaging ; Male

OBJECTIVETo retrospectively investigate the diagnosis and the outcome of Caroli's disease treated by surgical procedures.

METHODSThe clinical data of 68 patients with Caroli's disease treated by surgical procedures between 1996 and 2002 were reviewed, retrospectively.

RESULTSThe patients, with a M/F ratio of 1:1.35 and a mean age of 46, presented mainly with recurrent cholangitis. Of all the patients, 26 had a history of operation for cholelithiasis or cholangitis. On admission, the image investigations suggested that the lesions located at left lobe in 44 patients, right lobe in 9 patients, and whole liver in 15 patients. The coexisting cyst in common bile duct was found in 20 patients. The malignant transformation was found in 5 patients (8.8%). Hepatectomy was performed in 82.4% of patients, with a morbidity rate of 15.0% and mortality rate of 0 after the surgery. The long-term outcome of symptom-free in hepatectomy group was 90.2%, significantly higher than the 33.3% in non-hepatectomy group (P < 0.01) after a 3 to 10 years of follow-up.

CONCLUSIONSHepatectomy offers a curative procedure for local Caroli's disease, and liver transplantation is a good option for diffuse sufferers.

Adolescent ; Adult ; Aged ; Caroli Disease ; surgery ; Female ; Follow-Up Studies ; Hepatectomy ; Humans ; Liver Transplantation ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

To effectively guarantee quality of randomized controlld trial (RCT) of acupuncture and develop reasonable content and checklist of on-site quality control, influencing factors on quality of acupuncture RCT are analyzed and scientificity of quality control content and feasibility of on-site manipulation are put into overall consideration. Based on content and checklist of on-site quality control in National 11th Five-Year Plan Project Optimization of Comprehensive Treatment Plan for TCM in Prevention and Treatment of Serious Disease and Clinical Assessment on Generic Technology and Quality Control Research, it is proposed that on-site quality control of acupuncture RCT should be conducted with PICOST (patient, intervention, comparison, out come, site and time) as core, especially on quality control of interveners' skills and outcome assessment of blinding, and checklist of on-site quality control is developed to provide references for undertaking groups of the project.
Acupuncture Therapy ; standards ; China ; Humans ; Quality Control ; Randomized Controlled Trials as Topic ; standards ; Research Design ; standards

The growth inhibitory effects of D-glucosamine hydrochloride (GlcNH(2).HCl), D-glucosamine (GlcNH(2)) and N-acetyl glucosamine (NAG) on human hepatoma SMMC-7721 cells in vitro were investigated. The results showed that GlcNH(2).HCl and GlcNH(2) resulted in a concentration-dependent reduction in hepatoma cell growth as measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. This effect was accompanied by a marked increase in the proportion of S cells as analyzed by flow cytometry. In addition, human hepatoma SMMC-7721 cells treated with GlcNH(2).HCl resulted in the induction of apoptosis as assayed qualitatively by agarose gel electrophoresis. NAG could not inhibit the proliferation of SMMC-7721 cells. GlcNH(2).HCl exhibited antitumor activity against Sarcoma 180 in Kunming mice at dosage of 125-500 mg/kg, dose of 250 mg/kg being the best. GlcNH(2).HCl at dose of 250 mg/kg could enhance significantly the thymus index, and spleen index and could promote T lymphocyte proliferation induced by ConA. The antitumor effect of GlcNH(2).HCl is probably host-mediated and cytocidal.
Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA ; metabolism ; DNA Fragmentation ; drug effects ; Glucosamine ; analogs & derivatives ; therapeutic use ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Male ; Mice ; Sarcoma 180 ; drug therapy