Chromatography, Gas ; methods ; Chromatography, High Pressure Liquid ; methods ; Humans ; Hypnotics and Sedatives ; blood ; Pentobarbital ; blood ; Pharmaceutical Preparations ; blood ; Sensitivity and Specificity

Background The pathogenesiof age-related maculadegeneration (AMD) iassociated with the senility of human retinal pigmenepithelium (RPE) cells.Seeking drug to arresRPE cell senility iof significance fothe prevention and treatmenof AMD.Research showed thathe lycium barbarum polysaccharide (LBP) can delay senility,buitinfluence on RPE cell aging iunclear.Objective Thistudy wato discusthe protective effecand mechanism of LBP on RPE cell aging.MethodPorcine retinal neural epithelial layewaisolated,and photoreceptooutesegmen(POS) waextracted by density gradiencentrifugation and marked by FITC.The POwathen co-cultured with RPE cellin the medium containing 0.01,0.10 and 1.00 g/L LBP fo24 hours.The areof fluorescence,representing the amounof POphagocytosed by RPE cells,wameasured undethe fluorescenmicroscope to evaluate the influence of LBP on the phagocytifunction of RPE cells.The POS-induced RPE lipofuscin-uptake cell model waestablished by co-culturing human RPE cellwith porcine POfo3 weeks.The RPE-POco-culture cell model waincubated in medium containing 0.01,0.10 o1.00 g/L LBP,and the autofluorescence caused by lipofuscin up-taken into RPE cellwadetected with flow cytometry.cell counting kiwaused to assescell proliferation and viability (value) 24,48 and 72 hourafteculturing.ResultPorcine POpresented athin rodundethe lighmicroscope and appeared abilayedisc-like structureundethe transmission electron microscope,and itFITC-labeled yellow-green autofluorescence waobserved undethe fluorescenmicroscope.No POwaup-taken into the RPE cellin the normal control group,buthe areof POphagocytosed by RPE cellwagradually enlarged with increasing doseof LBP,showing significandifference among the group(F =21.425,P =0.006).Compared with the POcontrol group,the phagocytosed areincreased avariouconcentrationof LBP+POgroup(P＜0.01).Flow cytometry showed thathe autofluorescence value in the POcontrol group wamore highethan thaof the normal control group.Athe LBP dose increased,the autofluorescence value in the RPE celldeclined gradually and iwaneathe normal value in the 1.00 g/L LBP+ POgroup.The rate of proliferation of the lipofuscin RPE cellvaried with the increase of doseof LBP with the maximal value in the normal RPE group and minimal value in the lipofuscin RPE group,and the rate of proliferation of the lipofuscin RPE cellascended with increasing doseof LBP until neathe normal value in the 1.00 g/L LBP + lipofuscin RPE cellgroup (P＞0.05).ConclusionLBP enhance the anti-aging effecof human RPE cellby strengthening the phagocytiability to POand the ability to remove lipofuscin and by heightening the proliferation of human RPE cells.

·AIM:To present a case of late onset bilateral keratectasis after laser in situ keratomileusis (LASIK) for myopia with rigid gas-permeable contact lenses with a brief review of literature on this subject.·METHODS:A 27-year-old woman underwent bilateral uneventful LASIK for moderate myopia. Preoperative cycloplegic refractions were -5.50/-0.50×50° right eye (OD) and - 4.50/-1.00×15° left eye (OS).Corneal pachymetry was 526μm OD and 541μm OS, Preoperative corneal topography was normal and did not reveal any keratoconus or forme fruste keratoconus.Following the creation of flaps with 160μm plates,ablations of 102μm OD and 86μm OS were performed,estimated to leave residual stromal beds of 264μm OD and 295μm OS.·RESULTS:Twenty-nine months postoperatively,the patient developed bilateral inferior keratectasia of -12.50/-4.00×160° OD and -6.00/- 4.25×125° OS.Visual acuity was reduced in both eyes;the central cornea had steepened; and pachymetry showed central corneal thinning.Keratectasia was diagnosed,and rigid contact lenses were fitted.Three years later,the patient achieved satisfactory visual acuity and all-day lens wear with minimal complications.·CONCLUSION:Late keratectasia may follow LASIK for low to moderate myopia despite a thorough preoperative work-up.Rigid contact lenses can offer a safe,reversible option for improving visual acuity in such patients by delaying or avoiding the need for intracorneal ring segments implanting or penetrating keratoplasty.

OBJECTIVETo investigate the effect of compound Coptidis Rhizoma capsule (CCRC) on unbalanced expression of renal tissue TGF-β1/BMP-7 and Smad signaling pathway in rats with early diabetic nephropathy (DN), and discuss CCRC's effect on DN rats with early diabetic nephropathy and its possible mechanism.

METHODDN model rats were established by injecting streptozotocin (STZ). The rats were randomly divided into seven groups: the normal group, the model group, the enalapril treatment group, the xiaoke pill treatment group and three CRCC treatment groups. They were orally administered once a day for five weeks. The fasting blood glucose (FBG), blood urea nitrogen (BUN), serum creatinine (Scr), insulin (Ins), 24 h urinary protein (24 h Upro) and 24 h urinary microalbumin (24 h UmAlb) were tested. The pathological changes in renal tissues were examined by optical microscopy. Immuno- histochemical measures were used to detect the expressions of TGF-β1, BMP-7, Smad2/3, Smad1/5, and Smad7 protein, and RT-PCR was used to detect TGF-β1 mRNA and BMP-7 mRNA in renal tissues.

RESULTCompared with model group, BUN, Scr, Ins, 24 h Upro and 24 h UmAlb levels decreased at different degrees in CCRC treatment groups; the abnormal pathomorphology in renal tissue was improved; immunohistochemistry results showed that the expression of TGF-β1 and Smad2/3 were reduced, while the expression of BMP-7, Smad1/5 and Smad7 increased in CRCC treatment groups; the expression of TGF-β1 mRNA were reduced, but the expression of BMP-7 mRNA had no obvious change in CRCC treatment groups.

CONCLUSIONCRCC can improve the early renal function, delay the progression of chronic renal pathology and maintain the dynamic balance of TGF-β1/BMP-7 expression in renal tissues of DN rats. The mechanism may be related to down-regulation of renal TGF-β1 and up-regulation of BMP-7 through Smad signaling pathway.

Animals ; Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Coptis ; chemistry ; Diabetic Nephropathies ; drug therapy ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Signal Transduction ; drug effects ; Smad Proteins ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Background Exophthalmos caused by brain-related diseases will result in the serious outcome if treated improperly.It will be helpful for the diagnosis of the disease correctly when signs and symptoms are taken into consideration.Objective This study was to analyze the cause and clinical manifestations of exophthalmos due to brain-related diseases.Methods The elinical data from 38 patients with exophthalmos caused by brain-related diseases was collected and analyzed retrospectively.The sex,age,cause,exophthalmos range,symptom and sign of patients were considered. Results The gender of 38 patients was 21 males(55.26％)and 17 females(44.74％),aged from 16 to 71 years old with the average age 37.21.Four types of brain-related diseases were associated with the exophthalmos of eyeballs,including benign tumor 65.79％(25/38)containing the meningioma 60.53％(23/38),chordoma 5.26％(2/38)；malignant tumor 5.26％(2/38)containing the malignant meningioma 2.63％(1/38),choroid tong one cases of papillary carcinoma 2.63％(1/38)；carotid-cavernous fistula 26.32％(10/38)；meningoencephalocele 2.63％(1/38).Exophthaimos ranged from 18 to 35 mm in the patients with the average value of 22.17 mm.In all of the 38 patients,the right eyes were 36.84％(14/38),and left eyes were 55.26％(14/38)and bilateral eyes 7.89％(3/38).Accompanying signs and symptoms of meningioma included long-term headache (65.22％),nausea and vomiting(21.74％).declined vision(17.39％),elevated intraocular pressure,visual field shrink and loss,etc..Accompanying signs and symptoms of carotid cavernous fistula were loss of vision,pulsatile exophthalmos,noise over the orbit,ocular limitation of movement,eyelid edema(80％),bulbar conjunctival congestion,episodes of eyelid,diplopia(80％),headache(60％). Conclusion Some brain diseases are often accompanied by exophthalmos.Exophthaimos can provide useful clues to the diagnosis of brain diseases.

OBJECTIVE: To observe the effects of intermedin preconditioning on hypoxic injury in rat's cardiac myocytes and to provide the hypothetical mechanism of sudden cardiac death in the field of forensic pathology. METHODS: The H9c2 cultured rat cardiac myocytes were randomly divided into control group, hypoxia group and IMD group. The myocardial cell viability, cellular ultrastructure, intracellular calcium concentration and apoptosis rate were determined by MTT assay, transmission electron microscopy, laser scanning confocal microscope and flow cytometry, respectively. RESULTS: Compared with the control group, cell viability obviously decreased with inner ultrastructure injury in the hypoxia group (P<0.05), while cell viability significantly increased in the IMD group by reducing the hypoxia injury of cardiac myocytes (P<0.05). Compared with the control group, [Ca2+]i (fluorescence intensity) and apoptosis rate significantly increased in the hypoxia group, but decreased in the IMD group (P<0.05). CONCLUSION IMD increases the cell survival rate and decreases the cell apoptosis inhibited by intracellular calcium overload from hypoxia. This finding may reveal the mechanism of protective effects of myocardial hypoxia, and provide a scientific basis for the identification sudden cardiac death.
Animals ; Apoptosis ; Calcium ; Cell Hypoxia ; Cell Survival ; Hypoxia ; Myocardial Ischemia ; Myocardium/cytology* ; Myocytes, Cardiac/physiology* ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effect of mesenchymal stem cells (MSC) on the activity of nuclear factor (NF)-?B in rats with myocardial infarction.Methods MSC were isolated from SD rats (120—150 g in weight).SD rats (180—200 g in weight) were subjected to MI by left coronary artery occlusion,and were allo- cated into three groups randomly:1)sham group (without ligation of the artery,n=8);2)injection of PBS solu- tion (n=8);3)injection of MSC (n=8).MSC or PBS solution was injected into myocardium from epicardium instantly after MI models were established.Four weeks after transplantation,cardiac function was evaluated u- sing physiological recorder.Western blot were performed to investigate the nuclear factor-? activity.The ex- pressions of tumor necrosis factor (TNF)-? and interleukin (IL)-6 were evaluated by RT-PCR and Western blot. Results 1)Mortality was 20%(2/10) in sham group,33.3%(4/12) in PBS group and 20%(2/10) in MSC group with no statistic differences between them(P=0.646).2) Hemodynamic measurements showed that MSC trans- plantation caused significant improvement in cardiac function,comparing with MI+PBS group.3) MSC inhibi- ted the activities of NF-?B in myocardium and down-regulated the expression of TNF-? and IL-6 in mRNA and protein level.Conclusion Transplantation of MSC improved cardiac function in MI rats,which may partly at- tribute to their immuno-inflammatory regulation mechanism.

This study was aimed to explore the effect of bortezomib and low concentration cytarabine (Ara-C) on proliferation and apoptosis in U937 cell line and its mechanism. The proliferation and apoptosis of U937 cells treated with bortezomib (10 nmol/L) and(or) Ara-C (50 nmol/L) were observed by cell count, cell morphology, flow cytometry and Western blot. The results showed that bortezomib and Ara-C alone inhibited U937 cell proliferation. The inhibitory effect was enhanced by combination of these two drugs, the inhibitory rates of U937 cell proliferation were (55.00 ± 2.81)% and (70.02 ± 3.33)% after treatment for 24 h and 48 h, respectively. Bortezomib and Ara-C synergistically induced apoptosis and decreased mitochondrial membrane potential in U937 cells. The percentage of Rhodamin123 positive cells was (38.70 ± 1.54)%. Bortezomib and Ara-C also synergistically induced activation of caspase-9, caspase-8 and caspase-3. It is concluded that the bortezomib and low concentration Ara-C synergistically induced apoptosis in U937 cells, mainly through mitochondrial pathway, and possibly through death receptor pathway.
Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Cycle ; drug effects ; Cytarabine ; pharmacology ; Humans ; Pyrazines ; pharmacology ; U937 Cells

OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism