BACKGROUND: Numerous studies have shown that nuclear factor-KB (NF-kB) was positively correlated with vascular endothelial growth factor (VEGF) in the cerebral hypoxia region. Thus, we assumed whether NF-kB in the pathway of hypoxia-inducible factor-1 α (HIF-1α) upregulating VEGF and plays a bridge effect.OBJECTIVE: To study the effects of NF-kB in HIF-1α pathway of increasing expression of VEGF using HIF-1α-modified neural stem cells (NSCs) as vectors.DESIGN, TIME AND SETTING: In vitro observation of cytology was conducted at the Neuroscience Institute, Jiamusi University from March to December 2008.MATERIALS: Wistar rats aged less than 24 hours of both genders were used.METHODS: NSCs were transfected after amplification of adenovirus vector HIF-1α (AdHIF-ia)-green fluorescent protein (GFP).Fluorescence detection was used to determine HIF-1α-GFP and blank vector Ad-GFP expression in NSCs. Protein was extracted from transfected NSCs, blank vector NSCs and normal NSCs, separately. Subsequently, NF-kB specific inhibitor pyrrolidine dithiocarbamate (PDTC) (50,150, 300 μmol/L) was added in HIF-1α-modified NSCs. Western blot analysis was used to determine changes in VEGF expression.MAIN OUTCOME MEASURES: The following parameters were measured: expression of HIF-1α, VEGF and NF-kB in NSCs in each group; VEGF expression in NSCs following treatment of NF-kB specific inhibitor.RESULTS: Gene expression was associated with MOI and transfected time following AdHIF-1α-GFP transfected with NSCs. After transfected AdHIF-1α-GFP in NSCs, HIF-1α, VEGF and NF-kB expression was positively correlated. Expression of VEGF was reduced in AdHIF-1α-GFP-modified NSCs following treatment of NF-kB inhibitor PDTC in a concentration-dependent fashion.There were significant differences in VEGF expression between each concentration group (P < 0.05-0.01).CONCLUSION: NF-kB in signaling pathway of HIF-1α-upregulated VEGF expression and played a bridging effect.

Objective To explore visual-spatial working memory deficits of patients with basal ganglia damage, based on which tried to provide the new method for detecting the injuries in basal ganglia. Methods Twenty-five patients with lesions in the basal ganglia and twenty-five healthy controls performed visual-spatial working memory tasks, including a face-recognition and a spatial delayed-response. Results For the basal ganglia damage group ,the correct rate of both visual- face ( 54.5 ± 9.6 ) % and visual-spatial ( 80.0 ± 11.7 ) % working memory tasks was significantly lower than that of the control group ( ( 64.3 ± 9.5 ) %, ( 93.6 ± 4.9) %, respectively) ,and the difference was statistically significant ( u= - 147.5,80.5, P＜0. 01 ). For the patients injured in the left basal ganglia, the correct rate of visual- face working memory (48.5 ± 5.4 )% was obviously lower than that of patients injured in the right basal ganglia ( 59.2 ± 9.8 ) %, and the difference was statistically significant ( u =25.5, P＜0. 01 ) ;but the difference of correct rate for the visual-space working memory was not statistically significant( u = 52.5, P＞ 0.05 ). In contrast to the controls, both the visual-face and visual-space working memory of the group with injuries in basal ganglia,had appeared to be disable. Conclusions The results confirmed that patients with lesions in basal ganglia had deficits of visual-spatial working memory,and that injuries either in the left or the right basal ganglia can probably cause the shiftiness of cognitive function. Therefore, the injuries in basal ganglia can be detected by the visual-spatial working memory tests.

Objective To estimate the effect of the NO on the medial amygdaloid nucleus(Me), we studied the origins of the NOS positive terminals in the Me. Methods Noergic afferent projection to Me was identified by a combined NADPH-d histochemical staning and retrograde CTb immunocytochemical method after microinjecting CTb into Me. Results The double labeled of neurons (NOS and CTb) were located in dorsal raphe nucleus, locus ceruleus, basolateral amygdaloid nucleus, parabrachial nucleus, ventrolateral part of periaqueductal gray.Conclution The NADPH-d positive terminals in the Me originates from the aforementioned nucleus, and may relate to the function of the Me.

The basement membrane (BM) is crucial in regulating the physical and biological activities of esophageal epithelial cells which attach to the underlying BM. In order to simulate the natural construction of BM, we prepared the fibrous scaffolds using biodegradable polylactide (PLA) and silk fibroin (SF) as the materials via electrospinning technology. BM's proteins containing collagen (IV), laminin, entactin and proteoglycan were extracted from porcine esophagus and coated on the eletrospun fibers. Morphology, mechanical strength, biodegradability and cytocompatibility of the coated and uncoated scaffolds were tested and evaluated using scanning electron micrography, mechanical test system, immunofluorescence assay and western blotting with CK14 as the primary antibody. The fibrous scaffold PLA or PLA/SF, generated from the present protocol had good formation and mechanical and biodegradable properties. After coating with BM's proteins, the scaffold could enhance the growth and differentiation of esophageal epithelial cells, which would contribute to remodel and regenerate the tissue engineered epithelium and further contribute to engineer the whole esophagus in future.
Absorbable Implants ; Basement Membrane ; Biocompatible Materials ; chemistry ; Epithelium ; Esophagus ; physiology ; Fibroins ; chemistry ; Humans ; Nanostructures ; chemistry ; Polyesters ; chemistry ; Regeneration ; physiology ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

Objective To investigate the role of mitogen-activated protein kinases (MAPK)/extracellular signal regulated kinase1/2 (ERK1/2) signal transduction pathway in regulating the expression of aquaporin 3 (AQP3) in human amnion epithelial cells.Methods Primary cell culture of human amnion epithelial cells deriving from amnion of term pregnancy with normal amniotic fluid volume (AFV) or isolated oligohydramnios was conducted in the Second Affiliated Hospital of Wenzhou Medical College from January to November 2011.Either group included 10 elective cesarean cases.The primarily cultured cells were treated with different concentrations (0,5,10,20 and 40 mol/L) of ERK inhibitors (U0126) for 12 h,and then the optimal concentration of U0126 which resulted in the lowest expression of phospho~ERK1/2 (p-ERK1/2) was added for different durations(0,2,6,12 and 24 h).Immunocytochemistry was used to detect the localization of AQP3 and Western blot analysis was used to examine the expression of total ERK1/2,p-ERK1/2 and AQP3 in human amnion epithelial cells.Statistical analysis was performed by t-test and one-way ANOVA.Results (1) Compared with those in normal AFV group,p-ERK1/2 and AQP3 expression in human amnion epithelium cells decreased in oligohydramnios group,respectively (p-ERK1/2:3.46 ± 0.33 and 2.46±0.25;AQP3:2.34 ± 0.18 and 1.56±0.10,t=9.243 and 13.292,P＜0.01).(2) In oligohydramnios groups,after treated with different concentrations of U0126,the expressions of total ERK1/2 did not change (F=0.365,P＞0.05).The expression of p-ERK1/2 and AQP3 in 5,10,20,40 μmol/L U0126 (p-ERK1/2:0.96±0.16,1.12±0.13,0.98±0.17 and 1.02±0.26; AQP3:1.10±0.09,1.12±0.08,1.13±0.06 and 1.11±0.06) were all significantly lower than those in 0 μmol/LU0126 group (p-ERK1/2:2.46±0.25; AQP3:1.56±0.10,P＜0.05).However,the expression of p-ERK1/2 and AQP3 showed no significant difference among 5,10,20,and 40μmol/L U0126 groups (P＞0.05).The optimal concentration of U0126 was 5 μmol/L.After treated with 5 μmol/L U0126 for 2 h,the expressions of p-ERK1/2 and AQP3 (1.27±0.29 and 1.44±0.12)were lower than those after treated for 0 h (2.55±0.12 and 2.15±0.09,P＜0.05).However,there was no significant difference among groups treated for 2,6,12 and 24 h.Therefore,the optimal treatment time was 2 h.(3) The expression of AQP3 was distributed in both cell membrane and cytoplasm in amnion epithelial cells with normal amniotic fluid volume or isolated oligohydramnios,but mainly in cytoplasm.U0126 did not change the localization of AQP3 expression.Conclusions U0126 inhibits the phosphorylation of ERK1/2 and expression of AQP3 of women with oligohydramnios,indicating that the MAPK/ERK1/2 signal transduction pathway might regulate the expression of AQP3 in human amnion epithelial cells,and therefore affect the balance of amniotic fluid volume.

BACKGROUND:Polyurethane has good mechanical and physical characteristics and is extensively used in
clinical and experimental studies, but its hydrophobicity and histocompatibility are not ideal, which limits its use in tissue engineering as a biomaterial scaffold to some extents.
OBJECTIVE:To observe the hydrophilicity of polyurethane membrane grafted with silk fibroin and glutin and its compatibility with human hypopharyngeal cel s.
METHODS:The changes in hydrophilicity of polyurethane membrane grafted with silk fibroin and glutin were detected by contact angle measurements. Human hypopharyngeal fibroblasts were cultured in vitro on
polyurethane membrane, silk fibroin-polyurethane membrane, glutin-polyurethane membrane and tissue culture plate. Cel compatibility was compared using cytometry and cel morphology obsevation.
RESULTS AND CONCLUSION:Hydrophilicity of silk fibroin-or glutin-polyurethane membranes significantly increased (P<0.01). The hydrophilicity of silk fibroin-polyurethane membrane was higher than that of
glutin-polyurethane membrane (P<0.01). The number of cel s on the tissue culture plate was the most. The number of human hypopharyngeal fibroblasts on the silk fibroin-or glutin-polyurethane membrane was higher
than that on the polyurethane membrane, especial y on the silk fibroin-polyurethane membrane. These suggested that hydrophilicity and cel compatibility of silk fibroin-or glutin-polyurethane membrane were elevated.

Objective To observe the therapeutic effect of Radix Astragali,Radix Rehmanniae combined with glucocorticoid(GC) in treating patients with systemic lupus erythematosus(SLE) and its influence on some experimental indexes.Methods 72 cases of SLE were randomly recruited into the integrated traditional Chinese and western medicine treated group (treated group) and the western medicine treated group (control group).The control group was treated with prednisone and cyclophosphamide,while the treated group was treated with Radix Astragali and Radix Rehmanniae on the basis of the control group.Both groups had been treated for 6 months as a therapy course.The use of prednisone dosage were recorded and its influence on indexes of clinical and laboratory value were observed.Results 27 SLE patients in the treated group and 34 in the control group were completely observed.The dosages of prednisone in both groups were decreased gradually after the state of disease had been relieved.The predrisone dosage in the treated group was lower than that in the control group in the therapy course(15.41 ±7.65)mg vs (17.25±8.83)mg,but without significant difference (t=0.85,P＞0.05).The total decreased prednisone dosage in the treated group (50.12±12.53 mg) was slightly higher than that in control group(48.96± 13.71) mg (t=0.34,P＞0.05).The ratio of cases who had to add prednisone for aggravating disease in treated group(15.92％) was less than that in the control group (38.24％),the difference was significant (x2=3.24,P＜0.05).The incidence rate of cushing syndrome,infection,diseases of digestive,cardiovascular anomalies in the treated group was slightly less than the control group (x2=0.42,0.98,0.01,0.03respectivly,P＞0.05),but the number of insomnia and hot flush were significantly larger in the control group than the treatment group (x2=4.24,5.93,P＜0.05).No difference about the SLEDAI,ESR,CRP,Blood Complement3,Blood imune globinG was found between the two groups (t=2.16,1.04,1.18,0.26,1.61 respectively,P＞0.05),and 24 hours urinary protein count decreased significantly in the treated group (1.06 ± 0.12) g/L,compared with control group (1.42 ± 0.54) g/L,(t =3.78,P ＜ 0.05 ).Conclusion Radix Astragal,Radix Rehmanniae combined with conventional treatment of western medicine could withdraw corticosteroid smoothly,relieve symptoms and alleviate some side-effects of western medicine.

AIM: To improve the prevent efficacy of peptide p277 in autoimmune diabetes. METHODS: The recombinant expression plasmid pET28-Hsp65-6×p277 was constructed by inserting 6×p277 which were amplified by PCR into the vector pET28-Hsp65. The plasmid pET28-Hsp65-6×p277 was transformed into E.coli BL21 (DE3) and the fusion protein (Hsp65-6×p277) was expressed effectively as soluble protein after inducing by lactose. The fusion protein was purified and then used to immunize 4-week old female NOD mice with three times of i.n. inoculations in the absence of adjuvants. Serum samples from the immunized mice were collected at monthly interval. The concentrations of blood glucose and antibodies were measured by automatic analyzer. RESULTS: Administration with the Hsp65-6×p277 to NOD mice could prevent the development of diabetes. CONCLUSION: The fusion protein Hsp65-6×p277 might be further developed to a vaccine against insulin-dependent diabetes mellitus.

Objective To observe the effects of lower limb motion control training using virtual games combined with core stability training (CST) on the lower limb motor function of hemiplegic patients after stroke.Methods Sixty hemiplegic stroke patients were divided into an observation group and a control group (n =30 in each) using a random number table.The control group was given conventional treatment and CST,while the observation group received lower limb motion control training using virtual games in addition to conventional treatment and CST.All of the patients were assessed using the Fugl-Meyer lower limb assessment (FMA),the Berg balance scale (BBS),the modified Barthel index (MBI),10 metre maximum walking speed (MWS) and functional ambulation categorization (FAC)before and after 8 weeks of treatment.Results Before the intervention there was no significant difference between the two groups in any of the measurements.After the treatment,however,the average FMA,BBS,MBI,MWS and FAC in the observation group were significantly better than those before treatment and those in the control group.Conclusions Lower limb motion control training using virtual games combined with core stability training can improve effectively the lower limb motor function of hemiplegic patients after stroke.Such a combination is worth applying in clinical practice.

BACKGROUND:Bacterial biofilm is the main cause of the infection of the prosthesis.In vitro experiments confirmed that hypertonic sodium chloride and ethanol can apparently promote the formation of staphylococcal biofilms. There are no reports on the effects of ethanol and hypertonic environment surrounding the prosthesis on the formation of biofilms.
OBJECTIVE: To evaluate the effects of different environment factors surrounding the prosthesis on the growth of Staphylococcus epidermidis and bacterial biofilm formation after replacement.
METHODS: White rabbit models infected with Staphylococcus epidermidis on the prosthesis were established, and were randomly divided into hypertonic sodium chloride, ethanol and control groups (n=15). The bacteria were injected with 0.1 mL 4% sodium chloride and 4% ethanol into the knee of rabbits in the hypertonic sodium chloride and ethanol groups. The rabbits were injected with 0.1 mL 0.9% sodium chloride in the control group. Three rabbits were sacrificed at 2, 4, 6, 8 and 16 days after inoculated with bacteria. Synovial fluid, prosthesis and tissue surrounding infection were obtained. Bacterium was cultured to extract total RNA. The ica operon transcription levels were detected in the gene levels. Adhesion of bacteria on the surface of the prosthesis was observed using a scanning electron microscope. Tissues surrounding the prosthesis were observed using hematoxylin-eosin staining.
RESULTS AND CONCLUSION:Histological examination revealed that inflammatory cel infiltration was observed in al the rabbits at 4 days after injection. Colony formation was found at 16 days after injection. At 6 days after injection, inflammatory cel infiltration was observed in the ethanol and control groups. Scanning electron microscope showed that compared with the control group, the bacteria adhered to the prosthetic surface became more in the hypertonic sodium chloride and ethanol groups at 6, 8 and 16 days (P < 0.05). At 6, 8 and 16 days, the expression of icaA mRNA was significantly higher in the hypertonic sodium chloride and ethanol groups than in the control group (P < 0.05). These data showed that the environment factors could affect the growth of Staphylococcus epidermidis and bacterial biofilm formation.