Objective To test the role of a bioartificial kidney which consisted of a continuous vein-vein hemofiltration (CVVH) system and renal proximal tubule cells (PTCs)device, also called the renal tubule assist device (RAD),in endotoxin shock.Methods Female crossbred pigs 25 to 30 kg received with 3×1011 bacteria / kg body weight of E.coli intraperitoneally followed by treatment with a sham RAD without cells (n=7) or a RAD with cells (n=7). Data on blood pressure, cardiac output, renal blood flow and systemic marker of inflammation (IL-6) were measured.Results Cardiac outputs and renal blood flows were higher in the RAD group with cells than in the sham RAD group after E.coli infusion (P＜0.05). Plasma IL-6 levels were lower in the cell RAD group than in the sham RAD group after bacterial administration (P＜0.05). A significant difference on survival time was also observed between the two groups,with(9.07 ± 0.88) h in the cell RAD group vs. (5.10 ± 0.46)h in the sham RAD group, P＜ 0.01.Conclusion The bioartificial kidney containing PTCs can improve cardiovascular function,reduce cytokine levels,and prolong survival time.

The influence of L-arginine on endothelial nitric oxide synthase (eNOS) and cyclooxygenase 2 (COX2) was observed in experimental pulmonary thromboembolism and the action mechanism on pulmonary thromboembolism was explored. Wistar rats were randomly divided into control group, model group and treatment group. Pulmonary thromboembolism models were established by auto-blood back transfusion, and L-Arg 100 mg/kg was intraperitoneally injected after successful model preparation. The animals were sacrificed at 3 h, 1 day, 3 days and 7 days after embolism. Plasma NO, TXB2 and 6-Keto-PGFla were detected. The expression of eNOS and COX2 protein and mRNA in pulmonary tissues was detected by immunohistochemistry and RT-PCR respectively. The results showed that pulmonary thrombosis could be seen post pulmonary embolism and inflammatory reaction was significant. Plasma NO was decreased (P<0.01), and the levels of TXB2, 6-Keto-PGF1alpha and T/P ratio were all elevated. The expression of eNOS protein and mRNA in the pulmonary tissue was down-regulated (P<0.05), while that of COX2 protein and mRNA was upregulated (P<0.01). In treatment group, the level of NO was increased, the levels of TXB2 and T/P ratio were decreased, but the level of 6-Keto-PGF1alpha was increased. The expression of eNOS protein and mRNA in pulmonary tissue was upregulated (P<0.05), while that of COX2 protein and mRNA was down-regulated (P<0.05). In conclusion, L-arginine can educe the role of pulmonary tissue protection through up-regulating the expression of intra-pulmonary NOS and down -regulating COX2 in pulmonary thromboembolism.

Objective:To evaluate the role of nuclear receptor subfamily 1, group D, member 1/brain and muscle Arnt-like 1(Rev-erbα/Bmal1) signaling pathway in myocardial ischemia-reperfusion (I/R) injury in diabetic rats and its relationship with autophagy.Methods:SPF-grade adult male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-220 g, were used in this study.Type I diabetes mellitus was induced by intraperitoneal streptozotocin 60 mg/kg.The rats were continuously fed for 8 weeks after successful establishment of the model.Thirty rats with type 1 diabetes mellitus were divided into 3 groups by a random number table method: diabetic sham operation group (DS group, n=6), diabetic myocardial I/R group (DI/R group, n=12) and diabetic myocardial I/R plus Rev-erbα antagonist SR-8278 group (DI/R+ SR group, n=12). Eighteen non-diabetic rats were divided into 2 groups by a random number table method: non-diabetic sham operation group (NS group, n=6) and non-diabetic myocardial I/R group (NI/R group, n=12). The myocardial I/R model was established by ligation of the left anterior descending coronary artery for 30 min followed by 120-min reperfusion in anesthetized rats.SR-8278 2 mg/kg was intravenously injected via the femoral vein at 1 h before ischemia in group DI/R+ SR.Blood samples were collected from the carotid artery immediately after the end of reperfusion for determination of serum troponin I (cTnI), creatine kinase-MB (CK-MB) and lactic dehydrogenase (LDH) levels (by enzyme-linked immunosorbent assay). Then the rats were sacrificed, and myocardial tissues were obtained for determination of myocardial infarct size (by TTC method), expression of Rev-erbα and Bmal1 mRNA (by real-time polymerase chain reaction) and expression of Rev-erbα, Bmal1, microtubule-associated protein 1 light chain (LC3) Ⅱ and LC3Ⅰ (by Western blot) and for calculation of the ratio of LC3 Ⅱ/LC3Ⅰand the number of autophagosomes (with a transmission electron microscope). Results:Compared with group NS, the percentage of myocardial infarct size, serum levels of cTnI, CK-MB and LDH and the number of autophagosomes were significantly increased, the expression of Rev-erbα and its mRNA in myocardial tissues was up-regulated, the expression of Bmall and its mRNA was down-regulated, and the ratio of LC3 Ⅱ/LC3Ⅰwas increased in group NI/R, and serum levels of cTnI, CK-MB and LDH were increased, the number of autophagosomes was decreased, the expression of Rev-erbα and its mRNA in myocardial tissues was up-regulated, the expression of Bmall and its mRNA was down-regulated and the ratio of LC3 Ⅱ/LC3Ⅰwas decreased in group DS ( P<0.05). Compared with group NI/R, the percentage of myocardial infarct size and serum levels of cTnI, CK-MB and LDH were significantly increased, the number of autophagosomes was decreased, the expression of Rev-erbα and its mRNA in myocardial tissues was up-regulated, the expression of Bmall and its mRNA was down-regulated, and the ratio of LC3 Ⅱ/LC3Ⅰwas decreased in group DI/R ( P<0.05). Compared with group DS, the percentage of myocardial infarct size, serum levels of cTnI, CK-MB and LDH and the number of autophagosomes were were significantly increased, the expression of Rev-erbα and its mRNA in myocardial tissues was up-regulated, the expression of Bmall and its mRNA was down-regulated, and the ratio of LC3 Ⅱ/LC3Ⅰwas increased in group DI/R ( P<0.05). Compared with group DI/R, the percentage of myocardial infarct size, serum levels of cTnI, CK-MB and LDH and the number of autophagosomes were significantly decreased, the expression of Rev-erbα and its mRNA in myocardial tissues was down-regulated, the expression of Bmall and its mRNA was up-regulated, and the ratio of LC3 Ⅱ/LC3Ⅰwas increased in group DI/R+ SR ( P<0.05). Conclusion:Rev-erbα/BMAL1 signaling pathway is involved in the process of myocardial I/R injury by regulating cell autophagy in diabetic rats.

Objective To investigate the infection of Helicobacter pylori infection on the serum concentration of homocysteine (Hcy) and its relationship with coronary heart disease (CHD) .Methods 159 cases of patients with CHD were selected as the re‐search subjects .They were divided into two groups :infection group and non‐infection group ,according to the results of 14 C‐urea breathe test .And they were also divided into three groups :negative group ,mild infection group and severe infestation group ,accord‐ing to the severity of infection .The CHD patients infected Helicobacter pylori were divided into three groups:low risk group ,mod‐erate risk group and high risk group ,according to SYNTAX scores .The serum Hcy concentration was determined by cyclic enzy‐matic method .Results Comparing with non‐infection group ,the serum Hcy concentration significantly increased in infection group (P<0 .01) .With the aggravation of Helicobacter pylori infection ,the serum Hcy concentration increased .There were significant difference among negative group ,mild infection group and severe infestation group (P<0 .01) .In the CHD patients infected Heli‐cobacter pylori ,the serum Hcy concentration also increased with the aggravation of the severity of coronary lesion .And there were significant difference among low risk group ,moderate risk group and high risk group (P<0 .01) .Conclusion Helicobacter pylori plays a role in the incidence and development of CHD through increasing the serum Hcy concentration .

Background Research showed that exposure of 530 nm monochromatic light can induce myopia in animal,and retinal Müller cells participate in the formation of myopia.However,the effect and mechanism of retinal Müller cells during the formation of monochromatic light induced-myopia is below understood.Objective This study was to investigate biologic characteristics of rat retina Müller cells and the expression of cell factors in Müller cells after being illuminated by the 530 nm monochromatic light,and discuss the role of the retina Müller cells in myopia induced by monochromatic light.Methods Immortalized rat retinal Müller cells were cultured with DMEM containing 10％ fetal bovine serum in a self-made cell incubator with monochromatic light by adjusting luminance of 530 nm LED source.The cells were exposed to 125,250 and 500 lx luminance respectively for 6,12 and 24 hours,and the cells without light-irradiation were used as control.The growth of the cells under the different light time and different illuminations was described by MTT as the absorbance at the wavelength 570 nm (A570),and cell cycle analysis of Müller cells was performed by flow cytometry 48 hours after cultured,and the expression of transforming growth factor-beta 1 (TGF-β1),tyrosine hydroxylase(TH),inducible nitric oxide synthase(iNOS) and basic fibroblast growth factor(bFGF)in the cells were detected by reverse transcription PCR(RT-PCR),respectively.Results The Müller cells were uniform in size with polygonal shape and defined edges.No statistically significant difference was found in the A570 value in the cells of the 125 lx and 250 lx illuminated groups compared with the control group in various time points(P＞0.05).However,significant lowing was seen in the A570 value in the cells of the 500 lx illuminating for 12 hours and 24 hours in comparison with the control group (P =0.013,0.001).Compared with the control group,the ratio of the number between G2 and G1 phase was not significantly declined in 125 lx,250 lx illuminating for 48 hours (P =0.073,0.330),and the ratio in the 500 lx illuminating group was significantly lower than those in the 250 lx illuminated group and the control group (P =0.028,0.038).RT-PCR revealed that the expression of TGF-β1 mRNA in the cells was higher in the 250 lx illuminated group than that of the 500 lx illuminated group (P=0.006).The expression of iNOS mRNA was gradually upregulated in the 250 lx illuminated group compared with the control group (P =0.001),but that in the 500 lx illuminated group was downregulated (P =0.000).The expression of bFGF mRNA was raised in the 125 lx and 250 lx groups but reduced in the 500 lx group when compared with the control group(P=0.002,0.000,0.005).Also,the expression of TH mRNA was significantly increased in the 250 lx group(P=0.000),but decreased in the 500 lx group(P=0.000,P=0.001).Conclusions The monochromatic light of 530 nm can inhibit the growth of rat Müller cells and downregulate the expression of myopia-related cell factors and therefore exert effect in the formation of myopia.

Background Wearing contaclenincreasethe risk of infection of the cornea.Some studieshowed the gas-permeability of materialused foconstructing corneal contaclenione of the contributing factorrelated to corneal health.Objective Thistudy wato observe the in vitro adherence ability of differenbacterito rigid gas-permeable contaclense(RGP-CL) made with varioumaterials.MethodContaclensemade with hexafocon,enflufocon opolymethyl methacrylate (PMMA) were placed into Staphylococcuaureus,Staphylococcuepidermidis,oPseudomonaaeruginosbacterial suspension(0.5 MCF) fo24 hours.The strength of bacterial adherence watested and studied by the methyl thiazolyl tetrazolium (MTT) colorimetrimethod based on absorbance (value),and the vortex method waused to calculate the colony forming units.The bactericlump formation waexamined with scanning electron microscope (SEM).ResultMTcolorimetrimethod showed thathe adherence ability of Staphylococcuaureuto hexafocon (value) wasignificantly lowethan thato enflufocon and PMMA,respectively (q=7.379,8.207,P＜0.01),buno significandifference wafound in the adherence ability of Staphylococcuaureubetween enflufocon and PMM(q =0.828,P＞0.05).The adherence ability of Staphylococcuepidermidito XO and enflufocon walowethan thato PMM(q =14.000,12.800,P＜0.01),buno significandifference wafound between the adherence of Staphylococcuepidermidito hexafocon and enflufocon material (q =1.200,P＞0.05).There wano significandifference in the adherence ability of Pseudomonaaeruginosto all three material(F=2.155,P=0.138).The vortex method presented the colony forming unitof Staphylococcuaureuto hexafocon,enflufocon and PMMwith (37.9± 1.5)×106,(49.9±2.2)×106 and (67.4± 1.6)×106,respectively,with significandifference among them (F =206.240,P＜0.01),showing the lowesvalue in hexafocon,the highesvalue in PMMand middle value in enflufocon (q=11.650,28.640,16.990,P＜0.01),Moreover,colony forming uniof Staphylococcuepidermidito hexafocon,enflufocon and PMMwa(7.9 ± 1.3) × 106,(10.5 ± 1.5) × 106,(11.2 ±1.2) × 106,respectively.And thaof hexafocon walowethan one of the PMMmaterial (q =5.060,P＜0.05).No significandifference wafound between hexafocon and enflufocon nobetween hexafocon and PMM(q =3.290,1.770,P＞0.05).In addition,the resultthacorresponded to the vortex method were seen in the MTcolorimetriassay (F =0.232,P =0.799).SEM examination showed dispersed population of Staphylococcuaureuand Staphylococcuepidermidion the surfaceof hexafocon and enflufocon;while much more Staphylococcuaureuand Staphylococcuepidermidiadhered on the surface of PMMA,forming net-like appearance.Conversely,high numbeof Pseudomonaaeruginoswaseen on the surface of all three materials,withounoticeable differencein the bacterial shape and quantity on each of the material.ConclusionThe adherence ability of bacterito PMMistrongethan thaof hexafocon and enflufocon,and gas-permeable material of RGP-CL doenoimpacthe adherence ability of bacteria.

Objective To analyze the cause and treatment of the postoperative complications using one-stage posterior spinal osteotomy in the treatment of severe spinal deformity.Methods From September 2006 to May 2013,17 patients with severe spinal deformity (congenital scoliosis in 11 cases,congenital kyphosis in 4 cases,and congenital kyphoscoliosis in 2 cases) underwent one-stage posterior spinal osteotomy,including 5 males and 12 females with an average age of 22.6 (14-51) years.The preoperative mean coronal Cobb angle was 109° (85°-160°) while the mean sagittal Cobb angle was 104° (65°-152°).Two patients had neurological symptoms preoperatively whose spinal cord function was D,evaluated by ASIA classification.All patients were treated with pedicle subtration osteotomy and pedicle screw internal fixation,which SPO osteotomy 2 cases,PSO osteotomy 11 cases,VCR osteotomy 4 cases.Results There were 17 cases of complications in 147 patients,the complication rate was 11.6％(17/147).The causes were as followed,screw malposition in 2 cases,compromised by close of resected areas in 2 case,residual bone compression in 1 case,acute spinal cord injury in 2 cases,infection in 2 cases,broken stick or loosen hat in 3 cases,and superior mesenteric artery syndrome in 5 cases.Postoperative neurological complications occurred in 7 cases.Two cases with preoperative ASIA D became ASIA C,5 cases with normal nerve function became ASIA C in 2 cases and ASIA D in 3 cases.After surgical exploration,given Methylprednisolone and neurotrophic drugs,removal or changing of the internal fixation,anti-infection and symptomatic treatment,15 cases recovered completely and 2 cases improved partially.Conclusion One-stage posterior spinal osteotomy for severe spinal deformity is technical demanding and risky,and the postoperative complications are common.Appropriate operative procedure,close observation of sensation and motor function,timely surgical exploration and nerve decompression,and early brace wear are all required.

Background Light-induced retinal damage models vary as many influence factors,herein the modeling method is difficult to copy.It is necessary to establish the grading standardization of retinal damage after retinal light exposure.Objective This study was to improve the modeling method and establish a grading standardization for light-induced retinal damage in rat.Methods Twenty-four SPF 8-10 week-old male SD rats were randomly divided into 4 groups and 6 eyes for each group.The rats were exposed to light intense of 5000 lx for 1,2,3 hours respectively in 3 groups,and other 6 rats served as the normal group.Full-field light exposure experiment was performed for each individual rat separately,and an annular illumination box was used tO ensure the experimental rat moving in a single direction and exposing the right eye in 5000 lx light surrounding during experimental duration.Ganzfeid electroretinogram(ERG)was recorded from the experimental rats at the fifth day after light exposure,and the animals were then sacrificed for histopathology observation to evaluate the retinal thickness change.All procedures which involved animals adhered to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.Results After exposing to intensity light for 1,2,3 hours,the b-wave amplitudes of rod response,maximal mixed response,oscillatory potential in scotopic ERG as well as cone response,20 Hz flicker response of photopic ERG were significant declined as lapse of light exposure time(F=71.690,P=0.000；F=56.250,P=0.000；F=23.610,P=0.000：F=27.130,P=0.000；F=27.030,P=0.000)and lowed by 26.2％,52.5％,70.7％,24.4％,39.3％,58.1％respectively at the end of experiment.Meanwhile,the b-wave latencies of rod response,maximal mixed response in scotopic ERG as well as cone response of photopic ERG were evidently different among different groups (F=1.370,P=0.282；F：0.800,P=0.508；F=11.840,P=0.000；F=2.080,P=0.136).Light induced retinal damage located mainly at the temporal retina area.After intensity light exposure for 1,2,3 hours,the thickness of outer nuclear layer at the superior temporal retina attenuated by 11.3％,25.6％and 72.5％,respectively(P<0.05).A significant difference was seen in mean thickness of outer nuclear layer at superior temporal retina among different groups(F=410.27,P=0.000). Conclusion A standardized grading method for light-induced retinal damage is recommended.The continuous illumination in a intensity of 5000 Ix for 1,2,3 hours can induce the mild,moderate or severe retinal damage respectively at temporal retina.

Background Autologous and allograft renal transplantation exist some disadvantages of less donor source and rejection.As a scaffold of cell in tissue engineering,fibroin was determined to have a good biocompatibility.But whether the fibroin membrane can become a substitution for tissue defect is seldom reported.Objective This experiment aimed to investigate the feasibility of silk fibroin membrane in the rabbit eyelid reconstruction in situ.Methods A 4 mmx3 mm tarsi defect model was created on the upper eyelids of 18 healthy New Zealand white rabbits.The eyelid reconstruction in situ was performed with regenerated silk fibroin membrane material in the right upper eyelids (silk fibroin group ) and allogenic sclera material (sclera group ) on the upper eyelids of fellow eyes.The grafts were clinically examined for the evaluation of inflammation and implant exposure at the first,second and forth week after operation.The inflammation response and collagen distribution were examined by hematoxylin & eosin staining and Masson staining.Expression of basic fibroblast growth factor(bFGF) in the grafts was detected by immunohistochemistry,and ImagePro Plus software was used for statistical analysis.Results All eyelid defects showed a primary healing.The surface of palpebral conjunctival was smooth and the inflammation of ocular surface was mild.The eyelid margin in the sclera group was more notch than that in the silk fibroin group.Results of pathological examination revealed that the arrangement of collagen fibers in the sclera group was more disordered,but that in the silk fibroin group was regular.The expression level(A value) of b-FGF in the operative area in silk fibroin group were 0.027 67±0.004 69,0.051 73±0.008 72,0.058 72±0.006 88,and those in the sclera group were 0.056 48±0.009 14,0.072 83 ± 0.009 17 and 0.078 73 ±0.010 84 in 1,2,4 weeks after operation,showing statistically significant differences between two groups in various time points ( t =- 6.38,t =- 4.99,t =- 2.87,P ＜0.05 ).Conclusions Silk fibroin membrane can reconstruct the eyelid shape in situ with the less inflammation response and good biocompatibility.Silk fibroin membrane could be used to support the eyelid as a new tarsal repairing materials.

Background Pupillary light reflex has been widely used in the diagnosis and evaluation of visual system and nervous system diseases.However,in animal experiments,functional evaluation of the visual system and nervous system needs more advanced technology and are affected by many factors.Objective This study was to explore the use of the dynamic pupillometer in evaluating pupillary light reflex and to discuss the influence of brightness of stimulate on relevant curve parameters in C57BL/6 mouse.Methods Ten healthy SPF male C57BL/6 mice were collected in this experiment.White light of five luminance levels (2,8,32,128,256 cd/m2) was used to stimulate the mice following a 2-hour dark adaptation.The stimulation was given at the 60-second intervals,for a duration of 100 ms at every stimulation.An infrared camera and video capture card were used to capture digital images during the measuring process in a scotopic environment,at a speed of 60 frames per second.Measuring outcome was saved in the*.AVI format.A software that was developed by our group was used to determine pupil diameter and output pupillary light reflex curve offline.Pupil initial diameter (R1),constriction amplitude (CA),constriction velocity (CV),latency (T1),time for maximum velocity (T2),time for maximum constriction (T3),time for maximun con-striction to 10.1％ R1 re-dilation (RT)and re-dilation velocity (RV)were assessed,and the correlations between luminosity and measuring parameters were analyzed using the Spearman rank correlation.The use of animals followed the Regulations for thd Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results R1 values showed no statistically significant difference among the 5 different luminosity groups(F=1.117,P=0.361).A positive linear correlation was found between stimulating luminosity and CA(r=0.508,P＜ 0.01),but negative correlations were seen between stimulating luminosity and CV or RV (r=-0.625,-0.609,P＜0.01).T1 and T2 values in the 5 different luminosity groups were not statistically significant (F =0.202,P =0.936 ; F =1.584,P =0.195).The different levels of stimulating luminosity showed positive linear correlations with T3 and RT values (r =0.791,0.609,P＜ 0.01).Conclusions The dynamic pupillometer can quantitatively measure the pupillary light reflex of C57BL/6 mice.The pupillary light reflex dynamic curve parameters of mouse were affected by stimulus luminosity levels.These outcomes offer a basis for the application of the dynamic pupillometer system for measuring pupillary light reflex in animal models.