Background Herpetic stromal keratitis (HSK) is an immunopathologic eye disorder mediated by CD4+ T lymphocytes.Interleukin-9 (IL-9) is a cytokine linked to the process of many immune inflammatory diseases,but whether IL-9 is involved in the immunopathology of HSK remains unclear.Objective This study was to investigate the expression of IL-9 in murine cornea during the development of HSK and its relationship with the degree of HSK.Methods One hundred and eighty clean BALB/c mice were divided into the normal control group (20 mice) and experimental group (160 mice).HSK models were established by scratching on the surface of the cornea followed by inoculation of 1 × 106 plague forming unit(PFU) herpes simplex virus type 1 (HSV-1) KOS strain.Change of ocular surface was examined under the slit lamp biomicroscopy before and I day,3,5,7,8,10,14,21 days after inoculation of HSV-1.The corneas of mice were collected on the time points mentioned above.The relative expression level of IL-9 mRNA in the cornea of mice was detected by real-time PCR.Immunocytochemical localization of IL-9 protein in murine cornea was viewed by a confocal microscope after preparation of the corneal cryostat sections.All experimental manipulations were undertaken in accordance with the institutional guidelines for the care and use of laboratory animals.Results Punctiform,dendriform or geographic defect in corneal epithelium were seen 3 days and healing 6 days after inoculation of HSV-1.However,edema and opacity of the corneal stroma appeared 7 days and peaked 14 days following the inoculation.Real-time PCR assay showed that very little of IL-9 mRNA (A260/A280) was expressed in the cornea in the mice of the control group.But in 1 day,3,5,7,8,10 and 14 days,the relative level of IL-9 mRNA was 6.37±0.45,5.66±0.53,3.93±0.35,3.62±0.34,3.23±0.18,2.57±0.14,2.19±0.20,with a significant difference among the various time points (F=92.764,P=0.000),and IL-9 mRNA in 1 day,3,5,7,8,10 and 14 days after inoculation was significantly higher than before inoculation (P＜0.05);while no markedly difference was found in IL-9 mRNA expression between the 21-day group after inoculation and the normal control group (P =0.340).IL-9 protein was expressed in the epithelial layer,stromal layer and endothelial cell layer of the corneas,with a stronger green fluorescence in the HSK mice,but no expression of IL-9 protein was seen in the control mice.Conclusions HSK model can be successfully created by corneal scratching and inoculation of HSV-1 KOS strain in BALB/c mouse.IL-9 is involved in the pathogenesis and development of HSK mediated by immunoresponse.

Head and Neck Neoplasms ; surgery ; Humans ; Robotics

AIM: To summarize clinical characteristics and pathogenesis of iridocorneal endothelial syndrome ( ICE ) and investigate the treatment and prognosis.
METHODS:The clinical data of 12 cases (12 eyes) who received treatment in southwest hospital during Jun. 2007 to Feb. 2015 were analyzed retrospectively. The essential progressive atrophy of iris included 7 eyes, Chandler syndrome included 3 eyes, Congan - Reese syndrome included 2 eyes.
RESULTS: A total of 8 eyes were carried out once or multiple filtration surgery; 4 eyes were treated with glaucoma valve implantation. Postoperative follow- up time ranged from 15mo to 5y with the average of 30mo. Three months to 16mo after the surgery, the intraocular pressure of 4 patients were elevated again. Postoperative intraocular pressure was poorly controlled.
CONCLUSION:ICE syndrome is a group of clinically rare and serious eye disease. The excessive proliferation of ICE cells causes the existence of the corneal endothelial cells adhesion to the chamber angle and iris surface, which cause iris atrophy, secondary glaucoma, corneal endothelial decompensation. Currently, glaucoma filtration surgery and glaucoma valve implantation can only control intraocular pressure for several months, but the long-term prognosis is poor.

Background Recently,there were many studies on corneal innervations during mammalian development.However,there were fewer studies on discussing corneal innervations before and after mouse eye openings.Objective The present study was to investigate the change in the regulation of corneal nerve fiber length and density before and after mouse eye openings to offer a basis for clinical research in human.Methods Thirty SPF C57BL/6 mice were divided into postnatal 1 day(P1 d),P7 d,P13 d(1 day before eye opening),P14 d(eye halfopened),P17 d(1 day after eye opening)and P23 d(7 day after eye opening)groups,with 5 mice and 10 eyes for each group.Entire corneal stretches were prepared and immunostaining with an anti-neuron-specific β-Ⅲ tubulin antibody was performed to label the corneal nerve fibers.Confocal microscopic pictures from the corneal dorsal-nasal region (DN),dorsal-temporal(DT),ventral-nasal region(VN)and ventral-temporal(VT)were taken using Delta Vision Core.From these pictures,the mouse corneal area,total length and density of nerve fibers in the 4 regions were calculated.The use of the animals complied with Statement of ARVO.Results Corneal areas of P1 d,P7d,P13 d,P14 d,P17 d and P23 d mice were(0.404±0.007),(1.362±0.154),(1.573±0.080),(1.603±0.046),(1.847±0.052),(2.445±0.798)mm2,respectively ; the total lengths of nerve fibers were(3.718±1.044),(19.065±3.350),(23.687±0.907),(27.309±2.477),(31.989±3.976),(41.214±1.573)mm,respectively ; the densities of nerve fibers were(9.592±1.138),(14.506±1.908),(15.088±1.241),(16.772±1.897),(16.821±2.102),(17.660±1.216)mm/mm2,respectively,all showing significant increases with age(F =22.906,P =0.000 ; F =0.424,P =0.000 ; F =2.375,P=0.000).A positive correlation of the increasing corneal areas and increasing lengths of nerve fibers was found(r=0.983,P＜0.01).Nerve fiber densities in the four corneal regions significantly increased with age(DN region:F =0.159,P =0.000 ; DT region:F =2.1 72,P =0.001 ; VN region:F =1.998,P =0.000 ; VT region:F=2.352,P=0.000).From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference(t =0.589,P =0.572); and the corneal nerve fiber densities in the DT region,VN region and VT region decreased by 4.6％,5.5％ and 0.1％,respectively,without significant difference from P14 d to P17 d(t=0.549,P=0.596;t=0.701,P=0.501 ;t=-0.100,P=0.919).Conclusions The development of nerve fibers in the whole cornea or the four corneal regions is influenced by eye opening in mouse to various extents.From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference.From P14 d to P17 d,the corneal nerve fiber densities in the DT region,VN region and VT region decrease by 4.6％,5.5％ and 0.1％,respectively,without significant difference.Afterwards,the growth of nerve fibers increased in pace and the growth rate is recovered.

Optically pure α-substituted propanoic acids and their derivatives represent as an important kind of organic building blocks and key intermediates,which has been widely used in the synthesis of chiral drugs.Some of them have been used directly as nonsteroidal anti-inflammatory drugs (NSAIDs),such as ibuprofen,naproxen,ketoprofen and so on.Dihydroartemisinic acid (DHAA),the same structure as the α-substituted propanic acids,is a key intermediate for the synthesis of artemisinin,the most effective and current used anti-malarial drug.Therefore,the asymmetric synthesis of α-substituted propanoic acids is always a hot topic for chemical scientists.Asymmetric catalytic hydrogenation attracts more and more attentions because of its atom economy and efficiency.This dissertation will disclose the asymmetric synthesis of α-substituted propanoic acids using transition metal-complex as a chiral catalyst.

Objective To explore the occurrence of diaphragmatic muscle fatigue(DMF) in children with pneumonia and its relationship with respiratory failure.Methods One hundred and twenty-six cases of pneumonia(pneumonia group) and 60 healthy children(healthy control group) were selected,and 2K-V1 type intelligence electric impedance respirograph was used to measure diaphragmatic function of the children in both groups to find if the ribcage-abdomen motion was synchronic and to calculate M levels.The abdomen impedance values as X-axis were calculated,which were regarded,and the chest impedance values as Y-axis were calcaulated,which described two-dimensional chart of ribcage-abdomen motion curve and the ? angle which was between connection of maximum X value to the smallest X value and the X-axis were calculated.According to the changes of chest-abdominal curve in one-dimensional chart and the changes of M value and ? angle,the degree of the diaphragmatic muscle fatigue was judged.Results There were 98 cases of DMF in the 126 children with pneumonia,and 68 cases of which were type Ⅰ DMF,and 30 cases were type Ⅱ DMF.One-dimensional chart showed contradictory chest-abdominal respiratory motional curve,and two-dimensional chart moved in a clockwise direction.M value was(46.1?8.4)%,? angle was(136.7?12.0) degrees in children with type Ⅰ DMF;One-dimensional chart showed that the peak value of chest-abdominal respiratory motional curve dislocates,and two-dimensional chart moved in a counter-clockwise direction.M value was(17.2 ? 3.2)%,? angle was(48.2 ? 9.5) degrees in children with type Ⅱ DMF;the control group had M value(4.3 ? 1.0)%,? angle was(31.7 ? 5.2) degree.There were significant differences in M value and ? angle between type Ⅰ DMF children,type Ⅱ DMF children and healthy children(Pa

Objective To observe the anatomical and histological features of anterolateral ligament (ALL)in the knee of Chinese adults,so as to identify the existence of ALL and provide an anatomical foundation for clinical reconstruction.Methods Ten adult knee specimens were randomly selected to be dissected,and the femoral,tibial and meniscus attachment points of the ALL were observed.The length,width and thickness were measured using the vernier caliper after the dissection.Three specimens were subjected to histological staining in the end.Results (1)ALL originated from the lateral femoral condyle—the same point of the lateral collateral ligament femoral side or the distal-anterior side,with its body divided into two branches,located in the tibia and the lateral meniscus respectively.The starting point of tibial side ALL was located at the mid-point of Gerdy's tubercle to fibula head,below tibial cartilage edge,with the meniscus point located in the lateral meniscus anterior horn and body junction area.(2) The average length of ALL is 38.89 ± 4.67 mm.The width in the femur,tibial attachment point was fan-shaped spread connected with sclerotin,being the narrowest at the joint line.The width at the femur,tibial attachment point and the joint line was 8.49 ± 1.36 mm,8.15 ± 1.38 mm and 6.49 ± 1.09 mm respectively,with the thickness of 1.33 ± 0.38 mm.The distance from tibia attachment points to the Gerdy's tubercle,fibular head and tibia cartilage margin was 22.59 ± 3.04 mm,21.15 ± 2.78 mm and 5.76 ± 0.57 mm respectively.(3) HE staining showed that ALL was dense connective tissue consisting of parallel arranged collagen fibers,while S-100 staining indicated that ALL contained sensory motor nerve fibers.Conclusion ALL is independent of the joint capsule and originates from the femoral lateral condyle.Its body is divided into two branches,located in the tibia and the lateral meniscus respectively.

Objective To study the physical and chemical properties of silibinin; To lay foundation for optimal formulation design.Methods The high performance liquid chromatography method was used to detect the equilibrium solubility of silibininin in various solutions. The partition coefficients of silibinnin in the n-octalution-water and n-octalution-buffer solution systems were determined by shaking flask method. The destructive tests were carried out on silibinin.Results The equilibrium solubility of silibininin at 25℃ was 1.352 mg/L in water and the largest in acetonitrile, and increased in basic buffer solutions and after adding surfactants, of which sodium dodecyl benzene sulfonate was the strongest in solubilizing ability. The apparent oil-water partition coefficient of silibinnin was 61.39. Silibinnin was unstable in the acidic, basic, oxidizing and reducing solutions.Conclusion The experiment results can provide references for designing new preparations of silibinin.

Objective To investigate the effect of euphorbiasteroid on inducing the apoptosis of HL-60 cells and demonstrate whether the Fas/FasL signaling pathway is involved in the induction of apoptosis. Methods HL-60 cells were treated with dose of 2.5,10,40 μg/ml of euphorbiasteroid in vitro for 24 h respectively.After that,cell counting Kit-8 was used to detect cell proliferation.The morphology of HL-60 cells were observed under light and fluorescent microscopy.The early cell apoptosis was detected by using flow cytometry with Annexin Ⅴ-FITC /PI double staining.The expressions of Fas,FasL,caspase-8 and caspase-3 mRNA were analyzed by the method of RT-PCR.The activities of caspase-8 and caspase-3 were examined by chromatometry.Results Compared with 1640 control group,HL-60 cell proliferation was inhibited significant-ly by euphorbiasteroid.The inhibition rates were (34.9 ±3.7)%,(54.6 ±5.2)% and (61.3 ±4.3)%respectively.Moreover,HL-60 cells exhibited typical morphological features.Early cell apoptosis rates of HL-60 cells were (23.4 ±3.1)%,(35.7 ±4.3)% and (53.2 ±3.9)% respectively.Furthermore,the expressions of Fas,FasL,caspase-3 and caspase-8 mRNA were up-regulated significantly after euphorbiasteroid administration in a dose-dependent manner (P<0.01 ).After treated with euphorbiasteroid,the activities of caspase-8 and caspase-3 were significantly enhanced (P<0.01 ).Conclusion The up-regulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL-60 cells.

Objective To investigate the essential role of Toll-like receptor (TLR) signaling in dendritic cells in rejection responses of skin transplantation in mice.Method Immature wild-type BALB/c bone marrow dentritic cells (BMDCs) were stimulated by a TLR agonist,CpG,in the presence of ST2825,an inhibitor of key molecule myeloid differentiation factor 88 (MyD88) in TLR signal transduction,or in the absence for 24 h in vitro.Co-stimulatory molecules CD80 and CD86 were analyzed by flow cytometry.Wild-type C57BL/6 na ve T cells stained with CFSE,a fluorescent dye,were co-cultured with BALB/c DCs for 3 days in the presence or absence of ST2825.CD3 +/CFSE+cells were analyzed by flow cytometry.Minor antigen-mismatched (HY-mismatched) skin allograft model was employed in this study in which the donors were all males and the recipients were all females.The experimental groups were divided as follows:the control group,the donors and the recipients were all wide type BALB/c mice; the MyD88 group,the donors and the recipients were all MyD88-konckout BALB/c mice; the DCs group,the donors and the recipients were all MyD88-knockout BALB/c mice and the recipients were transfused with donor-sourced DCs after transplantation; the experimental group,the donors and the recipients were all MyD88-knockout BALB/c mice and the recipients were transfused with donor-sourced DCs pre-treated with ST2825after transplantation.The survival time of skin allografts was observed.Result ST2825 could dose dependently inhibit the high level expression of co-stimulatory molecules CD80 and CD86 induced with CpG.Similarly,and it could also dosedependently inhibit the allo-reactive T cell proliferation upon the stimulation with DCs.The survival time of every group was as follows:the control group (22.8 ±2.8) days vs.the MyD88 group (＞100 days) vs.the DCs group (9.7± 2) days vs.the experimental group (＞100 days,P＜0.05,vs.the DC group).Conclusion DCs may play an essential role in rejection responses of transplantation via TLR signaling.The inhibitor of TLR signaling,ST2825,could inhibit the activation of DCs and the biological functions,which might be contributed to the inhibitory effects of ST2825 on DCs maturation and indirect suppression on the proliferation of allo reactive T cells.