Adult ; Endometriosis ; complications ; metabolism ; Endometrium ; metabolism ; Female ; Humans ; Infertility, Female ; etiology ; metabolism ; Leptin ; metabolism

Background Experimental study showed that heparanase (HPA)is overexpressed in choroidal neovascularization,suggesting that it may play a role in the pathogenesis of angiogenesis.To certify HPA inhibitor suppress the formation and development of new blood vessel has an important significance for the treatment of choroidal neovascularization.Objective The aim of this study was to investigate the effect of HPA inhibitor on the proliferation of human umbilical vein vascular endothelial cell (UVEC) and the expression of HPA.Methods Hunan UVEC was primarily cultured and passaged and the third generation cells were used in the experiment.Phosphomannopentaose sulfate (PI-88) solution,a HPA inhibitor,was prepared with endothelial cell medium and the end concentrations were 400.00,200.00,100.00,50.00,25.00,12.50,6.25 mg/L respectively.The cells were treated with PI-88 solutions for 24,48 and 72 hours.The growth and proliferation of human UVEC were analyzed using MTT colorimetric assay at absorbance 570 nm.The expression of HPA in the cells was detected by immunochemistry in 48 hours after addition of PI-88.Results Cultured human UVEC showed the fusiform and polygon in shape.The A570 values of human CVEC were significantly different among various concentrations of PI-88 groups (F=2.721,P=0.053 ) and different action time (F=9.656,P =0.002).When PI-88 was administered for 24 hours,the A570 values of human UVEC were insignificantly altered in comparison with the one without PI-88 culture group (P＞0.05 ).However,in 72 hours after experiment,the A570 values were significantly declined as the PI-88 concentration was ＞50.00 mg/L ( P＜0.05 ).When PI-88 was administered for 48 and 72 hours,the A570 values of human UVEC were significantly higher than those of 24 hours in ＜50.00 mg/L groups (P＜0.01 ),but no statistical differences were seen in ＞100.00 mg/L groups among various time points (P＞0.05 ).HPA was intently expressed in the cytoplasm of human UVEC.However,at 48 hours after addition of ＞25.00 mg/L PI-88,the HPA expression was obviously weaker.Conclusions PI-88 can suppress the growth and proliferation of human UVEC at the dose-and time-dependent manner by downregulating the expression of HPA in the cells.

Background Diabetic retinopathy (DR) is one of the most important microvascular complications of diabetes,which has become one of the leading causes of blindness.Neovascularization is the main pathological manifestations of DR,but its mechanism is unknown.There is a clear need to investigate its pathogenesis which can offer potential therapeutic targets.Objective The aim of this study was to investigate the expression and distribution of visfatin and vascular endothelial growth factor (VEGF) in diabetic model rats.Methods This study was approved by Animal Ethic Committee of Inner Mongolia Medical University.Sixty SPF 8-week-old male SpragueDawley rats were randomized into the diabetic group and control group.The rats were housed under a condition that alternated between 12 hours of light and darkness,with free access to rat food and water.Diabetes was induced by intraperitoneal injection of 60 mg/kg (0.60 ml/100 g) of streptozotocin (STZ) and control rats received equivalent volume of buffer.The models were regarded as successful when blood glucose was ≥ 16.7 mmol/L.Rats were sacrificed 12 weeks after the injection of STZ and retinal specimens were prepared to detect the expression of visfatin and VEGF.Total retinal protein was isolated from the retinas of experimental and control eyes,and the expression of visfatin and VEGF was assessed by Western blot.Frozen cross sections of retinas of 5 μm thickness were used to perform double immunofluorescence staining with anti-visfatin and anti-VEGF antibodies.Results Mean body weight of the diabetic rats was (189.02±11.34) g and that of the control rats was (489.57 ± 14.48) g at 12 weeks post-injection,showing a significant difference between them (t =5.236,P =0.003).Mean blood glucose level was (29.25±3.86) mmol/L in the diabetic group and (5.32±1.01) mmol/L in the control group,demonstrating a significant difference (t =11.778,P =0.000).Double immunofluorescence staining showed reduced expression of visfatin and VEGF in the retinal nerve fibrous layer and glial cells in the control rats.A stronger staining for visfatin and VEGF was found in the various layers of the retina in the diabetic rats,with an expression level of visfatin (A value) of 346.26±41.23,which was considerably higher than that of the control group (102.07±65.01) (t =8.291,P =0.000) in 12 weeks after injection.Furthermore,the expression of VEGF in the retina was elevated in the diabetic group compared with the control group (A value) (415.88±92.15 vs.113.06±32.06) (t=10.067,P=0.000).Conclusions Visfatin might contribute to the pathologic progression of diabetic retinal,neovascularization and it might play a synergistic role with VEGF in the pathophysiology of DR.

Adolescent ; Humans ; Male ; Neurofibromatosis 1 ; Neurofibromatosis 2

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.

ObjectiveTo analysis the clinical efficacy of using lateral homodigital flaps based on digital artery perforator to repair the fingertip defects. MethodsFrom October 2008 to August 2010,nine patients with twelve fingertip defects,including 5 thumbs,2 index fingers,3 middle fingers,2 ring fingers,underwent repair with lateral homodigital flaps based on digital artery perforator.The size of the flaps ranged from 2.7 cm× 1.4 cm to 3.1 cm× 1.8 cm.The donor site were covered by skin graft. ResultsEleven flaps survived.One case met with partial necrosis.The follow-up time ranged from 3 to 6 months(average of 4.5 months).The finges had good appearance.Ten cases had gained full postoperative sensory recovery and the two-point discrimination was 4-Smm at 3 months after operation.ConclusionUsing the flaps pedicled with digital artery perforator is a feasible solution for treatment of fingertip defects.

AIM:To determine the contents of sinomenine and chelidonine in Tong’an Injection(Caulis sinomenii, Chelidonium majus Linn, etc). METHODS: HPLC was used. The conditions included the gradient elution with methanol-0.1% triethylamine. The detection wavelength was set at 280 nm. RESULTS: The calibration curve was linear in the range of 162-1 620 ?g for the sinomenine and the range of 35-350 ?g for chelidonine, respectively. The average recovery for sinomenine was 99.56% and the relative standard deviation was 0.41%(n=5). The average recovery for chelidonine was 99.46% and the relative standard deviation was 0.62% (n=5). CONCLUSION: The method is simple, rapid and specialty. It can be used for the determination of sinomenine and chelidonine in Tong’an Injection.

Using the high efficient copper-adsorbing yeast strain Y17 as absorbing material, the major affect factors including pH, original concentration of Cu2+, cell biomass, adsorption time and temperature were examined, and then the absorbing sites of the Y17 was determined. The results showed that the solution pH was the most dominate factor which affected the biosorption of Cu2+, the other affecting factors were the ini- tial concentration of Cu2+, the cell biomass added, and adsorption time, respectively; the temperature had lit- tle effect on the rate of biosorption. The orthogonal experiment showed that the optimal absorption condition was as follow: the solution pH was 5.0, the absorption time was 40 min, the cell biomass of Y17 added was 5.0 g/L, and the concentration of Cu2+ was 8 mmol/L; the highest adsorbing rate was up to 82.7% at this condition. Based on the results of different pretreatments and the desorption of Cu2+, the cell wall of Y17 was identified as the main place occurring boisorption process, and the -NH2 group, -COOH group on the surface of the yeast cells played an important role on the boisorption process.

Background Many studies and clinical trials of pharmacologic vitreolysis are already under way to try to improve vitreo-retinal surgery and to liquefy and detach the vitreous from the retina ultimately, including chondroitinase,hyaluronidase,dispase and plasmin. However, there has not been any report on purification of human plasminogen from cord blood plasma and inducing posterior vitreous detachment of the animal eye at present.Objective This study was designed to isolate and purify the production of human plasminogen (Plg) from cord blood plasma with ethanol precipitation and evaluate the efficacy of Plg in inducing posterior vitreous detachment (PVD).Methods Human Plg was Separated and purified from cord blood plasma by ethanol precipitation method. The protein band corresponding to Plg with molecular mass of 92 000 was revealed in SDS-PAGE and confirmed by MALDI-TOF and Mascot database. Anion-exchange chromatography and plasminogen activity assay kit were used to obtain purified Plg with biological activity. Twenty-five fresh pig eyes were enucleated and assigned to 5 groups and 5 eyes for each group. The normal eyes were used as control group. Balanced salt solution(BSS)of 0.1 ml was intravitreally group and standard substance group. All of the eyes were then incubatedfor 60 minutes under the 37 ℃. Retinal histopathology and ultrastructure were examined under the light microscopy, scanning electron microscopy ( SEM ) and transmission electron microscopy (TEM). Results The Plg with potential fibrinolytic activity was successfully extracted and purified from cord blood plasma by ethanol precipitation method. No posterior vitreous detachment (PVD) was seen in normal control group, BSS group and r-SK group following the intravitreal injection under the sem. However,PVD was demonstrated in r-SK+ Plg group and standard substance group under the SEM. The inner limiting membrane ( ILM ) and the retina were well preserved in all of the experimental eyes. No retinal morphology and ultrastructural abnormality were found under the light and SEM and TEM. Conclusion Ethanol precipitation is a feasible way to isolate and purify Plg from human cord blood plasma. Extracted Plg shows potential fibrinolytic intravitreal injection of Plg.

AIM:To investigate the expression of aryl hydrocarbon receptor (AhR) in atrial tissues of the patients with rheumatic heart disease (RHD), and the effects of AhR on rheumatic atrial fibrosis.METHODS:Right atrial specimens obtained from the patients with RHD requiring valve replacement surgery were divided into chronic atrial fibrillation (RHD+cAF, n=11) group and sinus rhythm (RHD+sinus rhythm, n=25) group.The patients with congenital heart disease (CHD) and sinus rhythm (CHD+sinus rhythm, n=12) who underwent heart surgery served as controls.The collagen volume fraction in the atrial specimens was examined by Masson`s trichrome staining.The protein expression and distribution of AhR, AhR nuclear translocator (ARNT) and CYP1A1 were detected by the methods of immunohistochemistry and Western blot.The mRNA expression of AhR, ARNT and CYP1A1 was detected by real-time fluorescence quantitative PCR.RESULTS:Compared with CHD+sinus rhythm group, the collagen content and the expression of AhR, ARNT and CYP1A1 were significantly increased in RHD+sinus rhythm group and RHD+cAF group.Compared with RHD+sinus rhythm group, the collagen content and the expression of AhR, ARNT and CYP1A1 were significantly increased in RHD+cAF group (P<0.05).CONCLUSION:The expression of AhR is correlated with the degree of fibrosis.The expression of AhR/ARNT/CYP1A1 is increased in atrial tissues of patients with RHD, suggesting that AhR/ARNT/CYP1A1 should be involved in atrial fibrosis of the patient with RHD.