Objective To explore the effect and safety of rituximab (RTX) in children with frequently relapsed nephrotic syndrome.Methods Twelve children (8 male and 4 female) with frequently relapse nephrotic syndrome were treated with RTX.They all showed resistance to various immunosuppressive agents and relapsed when steroid was reduced at 1.5-2.0 mg/kg.All immunosuppressive agents were stopped.Steroid was prescribed at doses of 2 mg/(kg · d),followed by alternate-day dosing (2 mg/kg) after proteinuria was negative for 5 days,and then tapered by 5 mg every 2 weeks until to low dose (＜ 0.5 mg/kg on alternate day) or discontinued.RTX was administered at a dose of 375 mg/m2 once every week for 3 weeks.Results The patients were followed up for 4-19 months [median (7.79 ± 3.00) months].Proteinuria turned negative on 1-14 d after first RTX treatment.Mean steroid dosages were significantly reduced than before treatment [alternate-day dosing (1.83 ± 1.02) mg/kg vs alternate-day dosing (0.34 ± 0.16) mg/kg,t =3.78,P =0.002].Gingival hyperplasia and Cushing appearance were significantly improved and steroid-induced glaucoma disappeared.The height was significantly increased compared with before treatment [(2.21 ± 1.40) cm/year vs (8.27 ±2.10) cm/year].Six cases had no relapse during follow-up.Another 6 cases relapsed in 5.5-19.0 months.All patients showed CD19 + B lymphocytes depletion after RTX treatment.However,CD19 + recovery could be seen in 5.5-12.0 months.Serum immunoglobulin did not significantly change.None of the patients was found with adverse events.Conclusions RTX can significantly reduce the dosage of steroid or even stop medication in children free of immunosuppressive agents,maintain remission for (7.79 ± 3.00) months,significantly improve the adverse reaction induced by steroid and immune-suppression.Thus,for children with frequently relapsed nephritic syndrome/steroid-dependent nephrotic syndrome who show resistance to immunosuppressive therapies,RTX treatment may be considered.

?AIM:To study the clinical value of non-steroidal anti-inflammatory drug in adjuvant treatment of intravitreal triamcinolone acetonide ( IVTA ) for macular edema caused by retinal vein occlusion ( RVO) .?METHODS: Forty - eight eyes in 48 patients were randomly divided into trial and control group ( 24 eyes each ) in this prospective study. In the trial group, additional pranoprofen drops was administered from 1d before IVTA to 30d after injection. Central foveal thickness ( CFT ) was measured with optical coherence tomography ( OCT ) . Available documents of best corrected visual acuity ( BCVA ) , CFT, intraocular pressure and complications pre- and post-injection at 3d, 1,2wk, 1 and 3mo were evaluated.?RESULTS: After IVTA, BCVA was improved in both groups at different levels; but there was no statistically significant between two groups at each time point ( P>0. 05). The CFT values were 629 ± 43μm vs 605 ± 57μm before IVTA in the trail vs control groups (P>0. 05). The values were 432±74μm vs 511±32μm (t=7. 533, P<0. 05), and 275±54μm vs 379±29μm (t=13. 212, P<0. 05) of the trial vs control groups at 1 and 3mo after IVTA, respectively. Ocular hypertension occurred in 5 eyes after injection in trail group, and was controlled with anti-glaucoma medication and one eye with filtration surgery. Progression of cataract was noted in 3 of 35 phakic eyes and cataract surgery was performed in 2 eyes at 4-12mo after injection in trail group. Progression of cataract was noted in 4 eyes and cataract surgery was performed in 2 eyes at 4- 12mo after injection in control group. No retinal detachment and endophthalmitis happened during the whole period of follow-up.?CONCLUSION: Application of non - steroidal anti -inflammatory eye drops in perioperative period can be useful to improve the outcome of IVTA for macular edema, which needs further evaluation.

Purpose To report the c1inicopatho1ogica1 characteristics,diagnosis and differentia1 diagnosis of extranoda1 fo11icu1ar den-dritic ce11 sarcoma( FDCS)of the pharyngea1 region. Methods The c1inica1 features,histopatho1ogica1 changes and immunohisto-chemica1 findings were ana1yzed in three cases of FDCS with review of the re1ated 1iterature. Results Case 1,a 70-year-o1d man pres-ented with the comp1aint of a pain1ess mass in pharyngea1 region accompanied by shortness of breath for the past 2 months. Case 2,a 40-year-o1d woman presented with the comp1aint of pharyngea1 foreign body sensation and b1oody sputum for the past 1 month. Case 3, a 38-year-o1d man presented with the comp1aint of intermittent epistaxis for the past 2 months. 3 cases showed simi1ar morpho1ogies:the neop1astic ce11s were ovoid to spind1e-shaped,with indistinct ce11 borders,dispersed granu1ar chromatin,and scattered sma11 nuc1e-o1i. Notab1y,severa1 nuc1ear inc1usions were identified,and rare binuc1ear and mu1tinuc1eated ce11s were a1so present. There were main1y 3 kinds of growth patterns in the tumors:diffuse sheets,fascic1es,and storiform arrangements admixed with sma11 1ymphocytes, which sometimes gathered into a mass. Immunohistochemica11y,tumor ce11s( 3/3 )were strong1y and diffuse1y positive for fo11icu1ar dendritic ce11 markers CD21,CD23 and CD35. Tumor ce11s(3/3)were a1so diffuse1y positive for fascin and D2-40. Some tumor ce11s (1/3)were diffuse1y positive for CXCL-13. Ki-67 pro1iferation index was estimated at 6%-20%. Conclusions Extranoda1 FDCS of the pharyngea1 region is rare and misdiagnosis is frequent1y made. A comprehensive eva1uation of c1inica1 manifestations,patho1ogic features and immunohistochemica1 findings are essentia1 for definitive diagnosis.

Aged ; Cell Dedifferentiation ; Chondrosarcoma ; pathology ; Humans ; Ribs ; pathology

Cognitive impairment is a common complication of diabetes. Hippocampus plays an important role in cognitive function. In hyperglycemia, synaptophysin, a crucial synaptic vesicle membrane protein in hippocampus neuron is found to be down-regulated. Recent evidences have shown that angiotensin IV can facilitate memory acquisition and recovery. However, whether it can also improve cognitive functions of diabetic rats with cognitive disorder, and the possible mechanisms are uncertain. Hence, the objectives of this study. Forty fi ve Sprague Dawley male rats were randomly divided into three groups: Control, diabetic group and diabetes with angiotensin IV treatment group. The cognitive functions, mainly learning and memory of the rats were evaluated using Morris water maze task. The synapses ultrastructure, relative mRNA concentrations and protein expression levels of synaptophysin in hippocampus CA1 area were estimated using transmission electron microscope, RT-PCR, immunohistochemistry and western blotting, respectively. Our study showed that in the diabetic rats with angiotensin IV treatment, the cognitive impairment as measured by Morris water maze task improved, the ultrastructure of synapses in hippocampus reversed, the relative mRNA concentrations and protein levels of synaptophysin in hippocampus signifi cantly increased, when compared with diabetic rats. We conclude that angiotensin IV plays an important role in improving cognitive function of diabetic rats. The possible mechanisms are up-regulating the expression of synaptophysin and normalizing the ultrastructure of synapses in hippocampus.

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.

Objective To determine the effects of fluoride on osteoclasts's quantity and bone resorption function in vitro and its mechanisms. Methods The osteoclasts and bone marrow stromal cells(BMSCs) isolated from long bone of new born rats were cultured respectively in TC199 medium (containing 10% fetal bovine serum) with fluoride. The osteoclasts were inoculated in 96-well culture plate and ivory slice, BMSCs were inoculated in 6- well culture plate, respectively, medium were changed after 2 hours incubation. They were divided into control group, low-dose fluoride, medium-dose fluoride and high-dose fluoride groups, the doses of sodium fluoride were 0,2.5 × 10-5,5.0 × 10-5,10.0 × 10-5 mol/L, respectively. Tartrate-resistant acid phosphatase(TRAP) staining positive cells were counted under light microscope after TRAP staining on the 2nd and the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue. The expression of receptor activator of NK-κβ ligand(RANKL) and osteoprotegerin(OPC) was detected by real-time fluorescence quantitative (337.5 ± 70.5), (447.5 ± 43.4), (472.9 ± 34.8), (475.3 ± 24.3)/well in the control group, the low-dose, mediumdose and high-dose fluoride groups, respectively. The differences were statistically significant between these groups and the control group (all P < 0.05). After in vitro culture for 5 days, the numbers of osteoclasts were (92.5 ± 22.1), (123.0 ± 26.4), (135.5 ± 22.2), (136.9 ± 23.0) per well in the control group, the low-dose, medium-dose and high-dose fluoride groups, respectively. The differences were statistically significant between these groups and the (0.088 ± 0.030), (0.100 ± 0.018), (0.152 ± 0.015), (0.242 ± 0.031 )mm2 per piece in the control group, the lowdose, medium-dose and high-dose fluoride groups, respectively. The values of medium-dose and high-dose fluoride BMSCs in the control group, the low-dose, medium-dose and high-dose fluoride groups were 100.00 ± 56.02, 144.95 ± 97.21,223.25 ± 184.48,193.98 ± 137.93, respectively. The values of medium-dose and high-dose fluoride groups were significantly higher than that of control group (all P < 0.05). Conclusions Fluoride can cause increase in the number of osteoclasts in vitro and promote their cell differentiation and bone resorption activity, which may be related to increased expression ratio of RANKL/OPG mRNA in BMSCs.

OBJECTIVETo obtain more information on DNA fingerprintings and to confirm the genetic relationships of ten wide populations of Pinellia tornata.

METHODCollect wide-type or cultivated P. tornata from ten major production areas nation-wide as material. Twelve combinations of EcoR I and Mse I primers were used to asses divergence among ten populations. The data were analyzed using unweighted pair-group method, based on arithmetic averages (UPGMA) bootstrap analysis. Cluster analyses were performed by using NTSYSpc software version 2.1. The alkaloid was extracted from P. ternate with chlorolform and determined in UV absorption spectroscop.

RESULT1,673 bands were applied with 12 combinations. These results of cluster were congruent with differences in total alkloids.

CONCLUSIONThe results of the study showed a potential application of AFLP fingerprinting for identification of P. ternata.

Alkaloids ; analysis ; China ; Cluster Analysis ; DNA Fingerprinting ; DNA, Plant ; genetics ; Phylogeny ; Pinellia ; chemistry ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Quality Control

Objective To determine the therapeutic effect of recombined rat insulin-like growth factory 1 gene and transforming growth factor beta-1(TGF-?_1)gene on anterior cruciate ligament transection(ACLT)- induced osteoarthritis-like changes in NZW rabbit knee joints.Methods Eighteen NZW rabbits were divided into 3 groups randomly after osteoanhritis was established by ACLT and another six rabbits were used as normal control group(group 1).Chondrocytes which had been transfected with IGF-1 gene,co-transfected with TGF-?_1 and IGF-1 gene(group 3,4)were injected into the rabbits knee joints.Experimental control group(group 2)only had ACLT bul was not transfected.After 4,8 weeks,rabbits were sacrificed and their joints were evaluated by morphological grades,histological examination,in situ hybridization examination,immunohistochemistry exami- nation,and transmission electron microscopy examination(TEM).Results The morphological grades showed that the normal control group had a very significant difference with the experimental control group(P

To obtain a number of penicillinases and degrade penicillin in milk by using the penicillinases,the gene encoding penicillinase was amplified by PCR from Bacillus cereus ATCC10987,cloned into pET28a(+) ,transformed into E. coli BL21;analysis of SDS-PAGE and penicillinase activity of the recombinant protein were done under induction of IPTG and the result showed that the maximum penicillinase activity reached 480 U/mL;the purity of penicillinase purified by Ni2+ Purification System was more than 90%;the immobilized penicillinases were obtained by sodium periodate method and the residual quantity of penicillin in milk(containing 0.5 U penicillin G/mL) was less than 4 ppb after degraded by the immobilized penicillinase.