BackgroundRetinitis pigmentosa (RP)is a group of progressive monogenic inheritance disease.Seldom epidemiology is performed to summarize the varied clinical phenotypes,especially some sporadic cases with untypical genetic history.ObjectiveThe aim of this survey was to investigate the clinical epidemiological characteristics and phenotype of sporadic RP.MethodsA prospective cohort study was designed.A survey of a series of clinically diagnosed sporadic primary RP patients was conducted at the Southwest Eye Hospital from July 2010 to November 2011.A total of 130 patients that matched the inclusion criteria were enrolled in this survey.Clinical ocular examinations and questionnaire surveys were given,including ophthalmoscopic examination,best corrective visual acuity( BCVA ),perimetry and Ganzfield electroretinogram (ERG)and color fundus photo.RP with different phenotypes were classified. ResultsA total of 130 sporadic RP patients were collected in this survey.Of them,66 were male and 64 were female with a mean age of (36.9±14.4) years.The average onset age of these subjects was (21.2±18.4) years.Seven (5.38％) patients had consanguineous marriage history,and 13 ( 10.00％ )patients had systemic disease.Forty-four (33.85％) patients had outdoor jobs,and 86 (66.15％ ) worked indoor.Eighty-nine patients had typical RP ( 68.5％ ),and the number of patients that developed central RP and sine pigmento RP were 16 ( 12.3％ ) and 16( 12.3％ ),respectively.An absence of a- and b-waves in full-field ERG wasdetected in 99 (76.15％ ) cases.The longest duration of night blindness was identified in typical RP patients and the lowest BCVA in central RP patients.ConclusionsThe age at first onset is early in sporadic RP.There are wide variations in different types of RP,but the ERG outcome is specific for all RP types.

Background Chondroitin sulphate proteoglycans (CSPGs) can cause the termination of ocular dominance plasticity in the visual cortex.Recently,protein tyrosine phosphatase σ (PTPσ) has been identified as a receptor that inhibits CSPGs.However,whether PTPσ and its downstream molecules participate in the reactivation of ocular dominance plasticity in adult visual cortex has not been studied.Objective The present study was to investigate the changes in the expression of the PTPσ,probabilistic neural networks (PNNs),and molecules downstream of PNN,such as N-cadherin/β-catenin,after the reactivation of adult visual cortical plasticity.Methods Fifty-four SPF Long Evans rats were grouped according to different postnatal week (PW) as the PW1 (6 rats),PW3 (6 rats),PW5 (6 rats),PW7 (24 rats),and PW9 (12 rats) groups,and the upper and lower eyelids were sutured in the 12 rats from the PW7 group for 14 days to establish the binocular plasticity reactivation models.Expression of PTPσ and PNNs in the rat visual cortex was detected using immunochemistry,and changes of PTPσ mRNA,N-cadherin mRNA and β-catenin mRNA expression in the rat visual cortex with plasticity reactivation were assessed by real-time fluorescence quantitative PCR (RT Q-PCR).The use of animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The expression level of PTPσ mRNA was significantly higher in the PW9 group than that of the binocular plasticity reactivation models and the PW7 group (t =1.965,3.526,P＜0.01).The staining of the rat visual cortex for PTPσwas localized to the cellular membrane,cytoplasm and axon.Cell densities of the PW9 group in the Ⅱ-Ⅲ layer,Ⅳ layer and Ⅴ-Ⅵ layer of the visual cortex were elevated in the PW9 rats compared with the PW7 rats (t =24.593,23.444,13.556,P＜0.01) and rats from the binocular plasticity reactivation model (t =44.111,43.000,16.556,P＜0.01).Cell densities for PNNs in the Ⅳ and Ⅴ-Ⅵ layers were significantly increased in the PW9 rats in comparison with the PW7 rats (t=1.926,P＜0.01 ;t=1.370,P＜0.05),but the cell density in the Ⅱ-Ⅱ layer has no statistical significance (t=0.889,P＞0.05).However,cell densities for PNNs in the Ⅱ-Ⅲ and Ⅳ layers in the binocular plasticity reactivation models were lower than those of the PW9 rats (t =2.556,4.585,P＜0.01).Compared with PW1 rats,the expression levels of the N-cadherin mRNA in the PW3,PW5,PW7,PW9 rats were lower (t =28.932,28.988,27.083,28.908,P＜0.01),but those in the PW7 rats were enhanced in comparison with the PW3 rats,PW5 rats and PW9 rats (t =1.848,1.904,1.825,P＜0.01).No significant difference was seen in the expression of the N-cadherin mRNA between the PW9 rats and rats from the binocular plasticity reactivation model (t =0.072,P＞0.05).A statistically significant increase was found in the β-catenin mRNA expression in the PW1 rats compared with the PW3,PW5,PW7 and PW9 rats (t =3.918,3.534,2.645,4.652,P＜ 0.0 1),as well as between rats from the binocular plasticity reactivation model and the PW9 rats (t =0.570,P＜0.01).Conclusions PTPr,PNNs and β-catenin are involved in the reactivation of ocular dominance plasticity in the adult visual cortex.

BACKGROUND: Studies on tissue-engineered vascular scaffold construction mostly focus on biodegradable scaffold and acellular allogenic or xenogenlc vascular scaffold. However, there are some problems to be urgently solved, such as control of degradable speed of biodegradable scaffold, and donor-sourced bacterial virus infecting recipients during the implantation of acellular natural vascular scaffold.OBJECTIVE: This study was designed to treat allogenic blood vessels by ultrahigh pressure in conjunction with nuclease washing (decellularization) to observe the decellularization effects and porcine endogenous retroviras (PERV) removal.DESIGN: A controlled observation.SETTING: National Cardiovascular Center, Japan.MATERIALS: This study was performed at the National Cardiovascular Center, Japan from April 2004 to April 2005.Young healthy male 1-3-month-old minipigs, weighing 3-5 kg, were provided by Japanese Farm. The protocol was performed in accordance with ethical guidelines for the use and care of animals. The main reagents and equipments used in the present study were as follows: Hoechst 33258 (Dojindo Laboratories, Kumamoto, Japan), ultrahigh pressure device (KOBELCO, Kobe Steel, Ltd, Japan), and PCR (GENEAMP PCR SYSTEM 9700).METHODS: Porcine descending aorta vessels were isolated under a sterile condition and treated by cold isostatic pressing (981 MPa, 4 ℃) for disruption of donor cells. The cell debris was digested by nuclease and washed out by phosphate buffered saline for vascular scaffold.MAIN OUTCOME MEASURES: After processing of decellularization by ultrahigh pressure treatment, vascular DNA levels were quantitatively determined by a fluorescent probe (Hoechst 33258); Removal of cell components from vascular tissue and retention of scaffold fibers were observed by a transmission electron microscope (JEM 100 cx); Scaffold ultrastructure was observed via a scanning electron microscope (JBM 5200); The morphological structure of vascular wall was observed via an optical microscope (100 augmentation) . All these were performed to evaluate the antigen-removal effects of decellularization by ultrahigh pressure treatment from histological, molecular biological, and immunohistochemical standpoints. Proviral DNA levels of acellular PERV were measured by PCR to evaluate the effects of decellularization by ultrahigh pressure treatment on killing PERV, a typical pathogenic microorganism.RESULTS: After decellularization by ultrahigh pressure treatment, the wavy structure of fibers was completely retained, and tissues were thoroughly cell free. Transmission electron microscope results demonstrated that collagen fibers and elastic fibers, but not cell components were detectable. Scanning electron microscope results demonstrated that only acellular scaffold was found. There was no PERV detected in the treated tissues. However, the PERV could not be inactivated in the tissues treated by surface active agent. Intravascular DNA levels significantly altered from (31.7±3.5 ) mg/L pre-decelhilarization by ultrahigh pressure treatment to (1.16±0.23) mg/L post- decellularization by ultrahigh pressure treatment(P＜0.01). Results demonstrated that decellularization by ultrahigh pressure treatment ridded of cellular nucleus and contents mostly.CONCLUSION: The study demonstrated that decellularization by ultrahigh pressure treatment could fundamentally rid cell components of scaffold, and concomitantly inactivate PERV successfully.

BACKGROUND:Mesenchymal stem cells (MSCs) exist in human tissues.Presently,cell source is single;culture method has great differences;obtained results are not consistent.Thus,it cannot verfy that isolated and cultured cells are identical calls,which is difficult to compare.OBJECTIVE:To compare the biological features of MSCs derived form bone marrow (BM),perpheral blood (PB) and cord blood (CB) under in vitro culture conditions.DESIGN,TIME AND SETTING:The cytological in vitro controlled study was performed at the Department of Hematology,Navy General Hospital of Chinese PLA from June 2007 to December 2008.MATERIALS:A total of 10 donors of hemopoietic stem cell transplantation at the Department of Hematology,Navy General Hospital of Chinese PLA were selected.MB and PB cells were obtained from the same donor,and cell volumes were respectively 20 mL and 2 mL.CB cells (30 mL) were obtained from healthy primipara at the Department of Obstetrics,Navy General Hospital of Chinese PLA.METHODS:MSCs were obtained from BM,PB and CB by Percoll density gradient + adherence method,and then incubated in DMEM/F12 medium containing 10% fetal bovine serum.When 80%-90% confluency,cells were digested in trypsin-EDTA and made into 5×10~8/L cell suspension as P_0.Above-described operation was performed as P_1,and the rest may be deduced by analogy as P_2-P_5.MAIN OUTCOME MEASURES:The following parameters were measured:cell growth morphology;results of Wright-Giemsa staining;results of cytochemistry;cell proliferation amount;cell surface markers using flow cytometry.RESULTS:Time of adherence,time to 50% confluency and time to 80% confluency of BMSCs were earlier comarped with the PBMSCs and UCMSCs.Adherent cells from BM grew in whirpool-like type,while CB and PB did not at 5-7 days.Majority of aderent cells from BM were fibroblast-like cells,and small parts were endothelioid cells.Aderent cells from PB and CB at the fifth generation contained more endothelioid cells and mononuclear and macrophage-like cells besides fibroblast-like cells.PAS stain,Sudan black B stein,alkaline phosphatase (AKP) staining of adherent cells from BM,PB and CB were negative from P_1 to P_5.Compared with P0 cells,number of BMMSCs till P5 was significantly more in PBMSCs and UCMSCs (P ＜ 0.05).Positive rates of CD29,CD44,CD90,CD71,CD105,CD166 and HLA-ABC were 55.9% 92.8% at P0 to P5,but ≤6% following BMMSCs were incubated;19.7%-33.4% at P0 to P5,but ≤10% following PBMSCs were incubated;35.4%-93.2% at P_0 to P_5,but ≤20% following CBMSCs were incubated.Positive rates of CD34,CD45 and HLA-DR were low in BM-,PB-and CB-MSCs.Positive rates of CD14 and CD31 were low in BMMSCs;12.1%-28.3% in PBMSCs,and 8.1%-21.3% in CBMSCs.CONCLUSION:MSCs can be attained from BM,PB and CB.Quantities of MSCs form BM are the highest,with single component,followed by CBMSCs and PBMSCs,with multiple components.

The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The chromatography condition was as follows: Kaseisorb LC ODS 2000 (4.6 mm x 150 mm, 5 min) column with the mobile phase of acetonitrile(A)-water( B) under gradient elution (0-5 min, 3%-8% A; 5-9 min, 8%-48% A; 9-23 min, 48%-62% A; 23-28 min, 62%-99% A); the flow rate was 1.0 mL x min(-1) with 10 microL injection volume; detection wavelength was 246 nm and temperature of column was 40 degrees C. The content of benzyl glucosinolate was in the range of 10.76-17.91 g x L(-1). The method is simple, accurate and good reproducibility which can be used for the determination of benzyl glucosinolate in Lepidium meyenii, effectively.
Chromatography, High Pressure Liquid ; methods ; Glucosinolates ; analysis ; Lepidium ; chemistry ; Plant Extracts ; analysis

To investigate the biological role of snake venom cystatin(sv-cystatin) in tumor progression, the cDNA of sv-cystatin amplified by PCR from pUC18-cystatin plasmid was cloned into methanol-inducible expression vector pPICZ?A. The linearized recombinant plasmid pPICZ?A-cystatin was transfered into Pichia pastoris, strain GS115 by electrophoration. Transfermants with phenotype Mut+ selected were identified by PCR analysis and induced in 1.0% methanol. The reombinant sv-cystatin protein was examined by SDS-PAGE, Western blot analysis. The molecular mass of expression product was about 14 kDa and approximately 16 mg/L of recombinant sv-cystatin was produced from one of GS115-cystatin transformants. The chromatography purified protein could reduce the activity of papain. The ability of B16F1 cells treated with recombinant sv-cystatin to invade the reconstituted basement membrane decreased significantly (P

Background:Acute-on-chronic liver failure( ACLF)is a commonly seen liver failure in China,and lacking an animal model that can effectively simulate the dynamic change of immune status of ACLF. Aims:To establish an animal model that can simulate dynamic change of immune status of ACLF by repeated administration of concanavalin A(ConA). Methods:Mice were randomly divided into normal control group and ConA repeated administration group. Mice in ConA repeated administration group were injected with ConA 15 mg/ kg through retrobulbar angular vein every 48 hours for 5 times,and mice in control group were injected with same volume of 0. 9% NaCl solution. Serum levels of IL-6,IL-10,IL- 12,TNF-α,IFN-γ,MCP-1 in peripheral blood were assessed by CBA assay,and the ratio of IL-10/ TNF-α was calculated. The expression of HLA-DR,number and proportion of CD4+ T cells and the expression of PD-1 of monocytes in peripheral blood were detected by flow cytometry. Results:Peripheral blood cytokines changed from predominated proinflammatory cytokines into predominated anti-inflammatory cytokines with the increasing in time of administration in ConA repeated administration group. Compared with control group,HLA-DR expression of monocytes in peripheral blood was significantly decreased(P <0. 05),number and proportion of CD4+ T cells were significantly decreased(P <0. 05), and PD-1 expression was significantly increased( P < 0. 05)in ConA repeated administration group. Conclusions:An animal model of ACLF immune status from systemic inflammatory response syndrome( SIRS) to compensatory antiinflammatory response syndrome(CARS)induced by repeated ConA stimulation is successfully established.

Reverse-genetic engineering of foot and mouth disease virus (FMDV) can improve the productivity, antigen matching, antigen stability, immune response ability, and biological safety of vaccines, so vaccine candidates with anticipated biological characteristics can be promptly achieved. Negative influence in taming of virulent strains can also be decreased or avoided. Reverse genetics not only make up for deficiencies like limitation of viral nature, low success rate, and time and energy consuming, but also realize more active designing of vaccines. Therefore, reverse genetics is significant in improving integral quality and efficiency of vaccines. In this review, we use FMDV vaccines as an example to summarize improvement in biological characteristics of virulent strains and provide a reference for related researches.
Animals ; Antibodies, Viral ; immunology ; Foot-and-Mouth Disease ; immunology ; prevention & control ; virology ; Foot-and-Mouth Disease Virus ; genetics ; immunology ; Reverse Genetics ; Viral Vaccines ; genetics ; immunology

OBJECTIVETo explore whether there is phenotypic modulation of mesangial cells in streptozotocin (STZ) induced diabetic rats and study the effect of Tujian Mixture (TJM) on it.

METHODSSD rats were divided into the normal control group (NC group, n = 8), the unilateral nephrectomized control group (QC group, n = 8), the STZ induced diabetes mellitus with unilateral nephrectomy model group (DM group, n = 8), the Valsartan treated group (VT group, n = 8) and the TJM treated group (ZY group, n = 9), rats in the latter two groups were modeled as in the DM group and treated with Valsartan (20 mg/kg.d) and TJM (20 g/kg.d) respectively for 12 weeks. The expression of alpha-smooth muscle actin (alpha-SMA) and transforming growth factor-beta 1 (TGF-beta 1) in rats glomeruli were observed by immunohistochemistry assay, and the ratio of alpha-SMA and TGF-beta 1 positive area/total glomerule tuft area (SMA/GT and TGF/GT) were analyzed using computer-assisted image analysis software.

RESULTSIn the NC and the QC groups, only trace of alpha-SMA positive staining was found. But there was prominant alpha-SMA positive staining in glomeruli of the DM group, with SMA/GT and TGF/GT increased significantly (P < 0.01), and marked increase of 24 hrs proteinuria excretion (P < 0.01). As compared with the DM group, the three indexes were all significantly lower in the VT and ZY groups (P < 0.01), and the lowering of proteinuria was more significant in the ZY group than that in the VT group (P < 0.01).

CONCLUSIONThe expression of alpha-SMA in glomeruli in STZ induced diabetic rats with unilateral nephrectomy is pronounced, indicating that phenotypic modulation of mesangial cells involvement in the pathogenesis of diabetic nephropathy. TJM and Valsartan can reduce 24 hrs proteinuria excretion, inhibit the phenotypic modulation of mesangial cells and the expression of TGF-beta 1 in glomeruli of diabetic rats, and the effect of TJM is more potent than that of Valsartan in lowering urinary protein excretion.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diabetic Nephropathies ; etiology ; metabolism ; pathology ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Glomerular Mesangium ; metabolism ; pathology ; Image Processing, Computer-Assisted ; Male ; Nephrectomy ; Phenotype ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; metabolism

OBJECTIVETo summarize experience of using screws and cement to rebuild tibial bone defect in primary total knee arthroplasty (TKA) and to discuss the relationship between the number of required screws and the severity of tibial bone defects.

METHODSFrom July 2009 to May 2015, 34 patients (40 knees) with varus knees underwent TKA, and the screw and cement technique was used to rebuild medial tibia plateau during operation. There were 8 males (8 knees) and 26 females (32 knees), and the average age was (65.00 +/- 7.25) years old (ranged,55 to 82 years old). One to 6 screws were used in each case. Extension stems were used in 2 cases (4 and 5 screws was used respectively). The area percentages of the bone defects measured as defect area/tibia plateau area, depth of each defect, the number of screws needed in each case, were all used to determine the relationship between the number of screws and the area percentage in certain depth of bone defect by statistic methods, as well as the relationship between screw number and defect depth.

RESULTSAll the patients were followed up and the average duration was 24 months (ranged, 1 to 72 months). The average preoperative HSS score was 43.33 +/- 6.11 (ranged, 32 to 51 scores). Whereas the average postoperative HSS score was 92.15 +/- 4.64 (ranged,83 to 96 scores). The preoperative individual scores including pain, function, activity, nuscle strength, flexion deformity and stability were all improved compared with preoperation,and the differences were statistically significant. All the patients received normal alignment postoperatively, femoraltibial angle was improved from (167.00 +/- 6.39) degrees preoperatively to (175.00 +/- 2.69) degrees postoperatively, the tibial angle was improved from (78.09 +/- 4.51) degrees preoperatively to (88.75 +/- 1.24) degrees postoperatively. Both area percentage and depth of bone defect in a fitting Ologistic model had a significant statistical relationship with the screw number, and a rectangular coordinate system could be formed according to the relationship.

CONCLUSIONScrews and cement technique is a simple, safe and convenient method to rebuild tibial bone defects in primary TKA and its short-term and midterm effect are both reliable. During opera- tion, according to the rectangular coordinate system, the screw number needed in the operation can be inferred form th area and depth of tibia defect, which could have a guiding function in surgery.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; instrumentation ; methods ; Bone Screws ; utilization ; Female ; Fracture Fixation, Internal ; Humans ; Knee Injuries ; surgery ; Knee Joint ; surgery ; Male ; Middle Aged ; Tibia ; surgery