A 1.8 kb HinfI fragment carrying the chitinase gene(chiA) from Serratia marcescens was isolated from the plasmid pLCHIA and was inserted into the SmaI site downstream of the strong Ptac promoter in the expression vector pKK223-3,yielding the plasmid pKChiA.The 2.1 kb PtacChiA fusion fragment was excised by BamHI from the plasmid pKChiA and was inserted into the single BamHI site present within the plasmid pMC71A,thus generating the plasmid pMChiA.High levels of chitinase were produced by the E.coli strains HB101 and JM105 carrying the plasmid pKChiA or the plasmid pMChiA,with the amount of production were higher 1～3 fold than that produced by the strains carrying the plasmid pLCHIA.

ObjectiveTo study the effect of hyperbaric oxygen therapy on the cholinergic neurons of hippocampus and praxiology of vascular dementia rats.MethodsThe vascular dementia models were made and divided into control and therapy group. After 30 days of HBO therapy, abilities of learning and memory of rats were tested by using the Morris water maze. Immunochemistry staining was used to observe the number of cholinergic neurons of CA1 subfield of hippocampus. ResultsAbilities of learning and memory of rats and the number of the neurons positive to ChAT like immunoreaction in the CA1 subfield of the hippocampus significantly increased in therapy group.ConclusionHyperbaric oxygen therapy is effective on the vascular dementia model rats.

OBJECTIVETo observe the effect and mechanisim of zoledronate sodium on periprosthetic osteolysis in rat induced by polyethylene particles.

METHODSTotal 30 adult male SD rats, weighting from 250 to 300 g, were selected and randomly divided into three groups: blank control group, model control group and zoledronate sodium group respectively, 10 animals for each group. No treatment was performed in the blank control group. In model control group and zoledronate sodium group, the modle of periprosthetic osteolysis in rats were made by implanting polyethylene particles and titanium rods into their right femurs. After operation, rats in zoledronate sodium group were administered with zoledronate sodium (0.1 mg/kg each week) through subcutaneous injection for 8 weeks, then the blood was obtained and all experimental animals were sacrificed to get the right femur specimens. The femur BMD, IL-1β, IL-6, TNF-α, serum TRACP5b and CTX-I were detected.

RESULTSCompared with the model control group, the femur BMD was increased, while IL-1β, IL-6 and TNF-α were all decreased in zoledronate sodium group; the serum TRACP5b and CTX-I level were both reduced in zoledronate sodium group.

CONCLUSIONThe zoledronate sodium could effectively inhibit periprosthetic osteolysis in rats induced by polyethylene particles, which might be realized by inhibiting the activity of osteoclasts and the expression of IL-1β, IL-6 and TNF-α. It provides a new method to treat periprosthetic osteolysis of the artificial joint prosthesis.

Animals ; Bone Density ; drug effects ; Collagen Type I ; analysis ; Cytokines ; analysis ; Diphosphonates ; therapeutic use ; Imidazoles ; therapeutic use ; Joint Prosthesis ; Male ; Osteolysis ; drug therapy ; Peptides ; analysis ; Polyethylene ; pharmacology ; Rats ; Rats, Sprague-Dawley

Adult ; Brain Neoplasms ; metabolism ; pathology ; surgery ; Female ; Ganglioglioma ; metabolism ; pathology ; surgery ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism

Eleven compounds were isolated and purified from the barks extract of Nothopanax delavayi and their structures were identified as serratagenic acid-3-O-alpha-L-arabinopyranosyl-28-O-beta-D-glucopyranosyl ester (1), serratagenic acid-3-0-alpha-L-arabi-nopyranosyl-28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester (2), serratagenic acid (3), serratagenic acid-3-O-alpha-L-arabinopyranoside (4), serratagenic acid-beta-O-beta-(2', 4'-O-diacetyl) -D-xylopyranosyl-28-O-[alpha-L-rhamnopy-ranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->46)-beta-D-glucopyranosyl] ester (5), serratagenic acid-3-O-alpha-(4'-O-acetyl)-L-arabino pyrano-syl-28-0- [-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester(6), serratagenic acid-3-O-alpha-(2'-O-acetyl)-L-arabinopyranosyl-28-O-[-alpha-L-rhamnopyranosyl- (1-->4) -beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester(7), serratagenic acid-3-0-beta-D-xylopyranosyl-28-O-[-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester (8), protocatechuic acid (9), ethyl caffeate (10) and caffeic anhydride (11) by physicochemical properties and spectroscopic data analysis. Among them, compounds 3-4 and 9-11 were firstly isolated from the genus Nothopanax, and compounds 5-8 were isolated from this plant for the first time.
Araliaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Bark ; chemistry

Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

Background Hypoxia is a crucial factor of neovascularization.Many researches found that stromal cell-derived factor-1 (SDF-1) and integrin-linked kinase (ILK) play an important role in the neovascular disease.However,effect of SDF-1 and ILK in eye neovascular disease is below understood.Objective The aim of this study was to investigate the effect of hypoxia on the expressions of SDF-1 and ILK in cultured retinal pigment epithelium(RPE) cells in vitro.Methods RPE tissue was isolated from 4-week-old C57BL/6 mouse and was digested and cultured in DMEM/F12 with 10％ fetal bovine serum (FBS).The cells with 80％ confluence were collected and passaged.The third generation of cells were identified with cytokeratin 18 (CK18) antibody by immunochemistry.The cells were inoculated at the density of 5×104 cells/ml to free-serum DMEM/F12 for 24 hours and then were cultured in regular medium in the normoxic control group.RPE cells were cultured for 1 hour and 3,6,12,24,48,72 hours with 200 μmol/L CoCl2 in the hypoxia group.Reverse transcription-PCR(RT-PCR) was used to evaluate the expressing change of SDF-1 mRNA and ILK mRNA in RPE cells,and Western blot was used to assay the expressing change of SDF-1 protein and ILK protein in RPE cells in different time points.The detected outcomes were represented as the ratio of target gene A value/β-actin A value.Results Cultured cells showed the polygon in shape with the black pigment granules in cytoplasm.Over 90％ cells were positive response for CK18.Expressions of the SDF-1 mRNA and ILK mRNA were increased in different time points after CoCl2 co-cultured(SDF-1 mRNA:F=281.875,P=0.000 ;ILK mRNA: F=187.566,P=0.000),with the highest expressing value in hypoxia at 12 hours.No significant change in the expression of SDF-1 mRNA and protein was found 1 hour after CoCl2 co-cultured,but expressions of SDF-1 mRNA and ILK mRNA were significantly higher in 3,6,12,24,48 and 72 hours than the normoxic control group(P＜0.01).The expressions of SDF-1 protein and ILK protein were gradually ascended with the time increase of CoCl2 co-culture,showing a significant difference among different time points(SDF-1: F=44.719,P =0.000 ; ILK: F =144.481,P =0.000),and the up-regulation of SDF-1 protein and ILK protein expression was seen mainly in 3,6,12,24,48 and 72 hours after CoCl2 co-cultured in comparison with the normoxic control group (P＜0.01).Conclusions SDF-1 and ILK are involved in the hypoxic response of RPE cells and may play a potential role in ischemic/hypoxic retinopathy.

Objective:To develop evaluation index system of Knowledge, Attitude, and Practice Model of esophageal speech training for Otolaryngology nurse, which can provide an initial tool to evaluate the effect of esophageal speech training for Otolaryngology nurse.Methods:Based on the Knowledge, Attitude, and Practice Model and literature research, the initial dimensions and items were developed. The evaluation index system was completed by Delphi technique that 23 experts were invited to evaluate the importance of the dimensions and items in the consulting round. Then the weight value of the index system was computed through analytic hierarchy.Results:For the first and second rounds of the study, respectively, the recovery rates were 78.26% and 100.00%, the expert authority were 0.85 and 0.86, and the Kendall′s W were 0.16 and 0.19 ( P<0.05). Finally, the index system contained 3 dimensions and 30 items. Conclusions:The evaluation index system has the possible scientificity and reliability, and has the feasible thoery and content. In the future, the evaluation index system can be used to evaluate the effect of esophageal speech training for Otolaryngology nurse through the clinical practice improvement.

BACKGROUND: The poor retention and survival of donor cells implanted into the myocardium limit the efficacy of cell therapy for myocardial infarction. Embedding cells in natural or synthetic biomaterials is a strategy to address this issue. OBJECTIVE: To explore the effects of bone marrow mesenchymal stem cells (BMSCs) encapsulated in hyaluronic acid (HA) hydrogel on cardiac function after myocardial infarction. METHODS:BMSCs from male Sprague-Dawley rats were isolated and cultured,and then HA-encapsulated BMSCs were cultured in vitro in the three-dimensional manner. A model of myocardial infarction was made by cutting the anterior descending artery of female Sprague-Dawley rats. After 1 week, the model rats were screened by ultrasonic testing and then eligible ones were randomly divided into four groups: PBS group (n=8), HA group (n=8), BMSCs group (n=29), and HA-encapsulated BMSCs group (n=29). At 1 week after modeling, the model rats underwent the secondary thoracotomy and the implants were injected into the marginal zone and infarcted region in corresponding groups. The survival rate and apoptosis of implanted cells were examined at post-injection day 1, week 1 and week 2 by RT-PCR and TUNEL respectively. At post-injection week 4, changes of cardiac microstructure and function were evaluated by histological examination and echocardiography. RESULTS AND CONCLUSION: Compared with the BMSCs group, HA hydrogel significantly enhanced the survival rate and reduced the apoptotic rate of BMSCs at post-injection day 1 and week 2 (both P < 0.05). At post-injection week 4, the HA+BMSCs combined treatment yielded the best recovery of cardiac function (P < 0.05). To conclude, HA hydrogel can act as a vehicle for BMSCs delivery and improve the beneficial effects of implanted BMSCs in early myocardial repair(within 2 weeks after infarction)via enhancing cell retention and survival.

OBJECTIVETo determine the mechanism of pseudo-tears of the lateral meniscus caused by the transverse geniculate ligament (TL) and the miniscofemoral ligament(MFL) and to investigate a method to differentiate pseudo-tears from true tear of the lateral meniscus.

METHODSForm June 2012 to February 2014, MR examinations of 72 knees (44 left knees and 28 right knees) without tear of the lateral meniscus verified by arthroscopy were performed in the sagittal and coronal plane. There were 41 males and 31 females in the group, with an average age of 33.7 years old (ranged from 25 to 61). The MR appearance of the TL and the MFL was carefully observed.

RESULTSThere existed fatty tissue in the gap between the TL and the anterior horn of the lateral meniscus and its central tendinous attachment. On the sagittal images, the fatty tissue formed a linear high-signal cleft between the TL and the anterior horn of the lateral meniscus. This might be mistaken as an oblique tear within the anterior horn of the lateral meniscus. It was called as pseudo-tears of the anterior horn of the lateral meniscus. In sagittal plane, the MFL was identified as a circle-like or short stick-like area of low signal intensity anterior or posterior to the posterior cruciateligament. Nevertheless, a belt-shaped area of low signal intensity from the posterior horn of the lateral meniscus to lateral facet of the medial femoral condyle was identified in the coronal plane. A linear area of high signal intensity between the MFL and the lateral meniscus was found in sagittal plane, which might be mistaken as an oblique tear within the posterior horn of the lateral meniscus. It was called pseudo-tears of the posterior horn of the lateral meniscus. The occurrence rate of the TL was 34.7% (25/72). The prevaleribe of pseudo-tears of the anterior horn of the lateral meniscus was 18 cases. The shape of the anterior horn of the lateral meniscus was regular, and the course of the pseudo-tears cleft was oblique. The occurrence rate of the MFL was 73.6% (53/72), which included the anterior MFL 23.6% (17/72), the posterior MFL 70.8% (51/72) and the two ligaments coexisted 16.7% (12/72). The prevalence of pseudo-tears of the posterior horn of the lateral meniscus was 25 cases. All observed pseudo-tears had either in posteroinferiorly oblique direction (19/25) or in vertical direction (6/25).

CONCLUSIONBased on the location and direction of pseudo-tears and observation in the continuous sagittal plane and the coronal plane, pseudo-tears is easily differentiated from the true tear of the lateral meniscus

Adult ; Female ; Humans ; Knee Injuries ; diagnostic imaging ; surgery ; Knee Joint ; diagnostic imaging ; surgery ; Magnetic Resonance Imaging ; Male ; Menisci, Tibial ; diagnostic imaging ; surgery ; Middle Aged ; Radiography ; Tibial Meniscus Injuries