Objective To observe the distribution and concentration of ~(125)I-nerve growth factor (NG-F) in rabbits' eyes after intravitreal injection and posterior juxtascleral injection.Methods Intravitreal injection(group A) and posterior juxtascleral injection (group B) were performed with the dosage of 30μg/100μl ~(125)I-NGF on left and right eyes in 45 white rabbits respectively.The γ-counts and the concentration of ~(125)I-NGF (%ID/g) of each ocular tissue was determined 15 and 30 minutes,and 1,3,6,12,24,and 48hours after injection.Results The ~(125)I-NGF diffusion in group A was faster in ocular content and ocular inner wall.The vitreous content of ~(125)I-NGF decreased gradually in group A,the curve changes in other eye tissues were normal.The concentration of ~(125)I-NGF reached the peak 3 hours after injection in aqueous humor,iris and ciliary body,retina,and choroids,but 6 hours after injection in sclera and 8 hours in cornea.The changes of concentration of ~(125)I-NGF in group B showed normal curve change.The peak time in group B were all 6 hours,in all the tissues except aqueous humor (3 hours).Except the high concentration in vitreous body caused by intravitreal injection,the concentration of ~(125)I-NGF in retina was the highest in group A.Conclusion Intravitreal injection of ~(125)I-NGF can gain higher concentration in each ocular tissue than posterior juxtaseleral injection,especially in retina.So intravitreal injection of NGF is a better ocular delivery method to treat the ocular fundus diseases.

Objective To investigate the prevalence,clinical traits and correlates of apathy in patients with Parkinson' s disease ( PD ).Methods Seventy-seven cases of PD patients and 40 health controls were recuited in the present study.Apathy was assessed using Lille Apathy Rating Scale(LARS) and related factors,such as motor function,cognition,depression,age,sex,education level,disease duration,levodopa equivalent doses (LED) and use of antipsychotic drugs were also evaluated.Four kinds of scales,Unified Parkinson' s Disease Rating Scale (UPDRS),Hoehn-Yahr staging,Montreal Cognitive Assessment (MoCA),Hamilton Depression Rating Scale (HAMD) were employed to evaluate the motor function,cogniton and depression,respectively.Results The prevalence of apathy and depression in the 77 PD patients were 49.4％ ( 38/77 ) and 46.8％ (36/77),respectively.Of the 77 PD patients,11 cases had apathy without depression (14.3％ ); 9 cases had depression without apathy( 11.7％ ) and 27 cases had apathy combined with depression( 35.1％ ).Among the PD patients with apathy,the cognitive type is the most common one,accounting for 86.8％ (33/38).There were significant statistical differences in education level,UPDRS- Ⅱ/Ⅲ scores,Hoehn-Yahr staging,MoCA and HAMD scores between apathy ( n =38 ) and no-apathy(n =39) PD patients(t =2.309,-3.144,-4.000,-3.217,2.649,-3.909,all P＜0.05).According to the unifactoral Logistic regression analysis of apathy correlates,UPDRS-Ⅱ/Ⅲ scores,HoehnYahr staging and HAMD scores were the risk factors for apathy while education level and MoCA were the protective factors for apathy.In the multiple Logistic regression analysis,HAMD scores,UPDRS-Ⅱ scores and education level were included.Conclusions The prevalence of apathy in PD patients was significantly higher than healthy controls.Apathy could be present without depression in PD patients,and was associated with education level,motor dysfunction degree,cognitive impairment and depression severity.

Objective To observe the clinical efficacy of bevacizumab intravitreal injection combined with laser photocoagulation for di-abetic retinopathy macular edema(DME). Methods 25 DME patients (15 patients, 24 eyes in control group and 10 patients,13 eyes in treatment group) were selected from January 2013 to November 2014 in our hospital. The control group received single Krypton laser (510 nm red and 483 nm yellow) grid photocoagulation or big“C” photocoagulation, while the treatment group received bevacizumab intrav-itreal injection after Krypton laser photocoagulation. The visual acuity were checked, and the macular retinal thickness was detected by opti-cal coherence tomography ( OCT) . Results The difference in preoperative visual acuity between the two groups was not statistically signifi-cant (P>0. 05). All visual acuity improved significantly (P<0. 05) after treatment and the treatment group got a better result compared to the control group(P <0. 05). The preoperative macular retinal thickness in treatment and control groups were respectively (510. 75 ± 61. 06)μm and(487. 47 ± 43. 41)μm, and the difference between them was not statistically significant (P>0. 05). And the macular retinal thickness were respectively ( 253. 41 ± 39. 12 )μm and ( 358. 81 ± 43. 24 )μm after treatment, which were all decreased than before (P<0. 05). The macular retinal thickness of the treatment group was better than that of the treatment group after treatment (P<0. 05). Conclusion Single Krypton laser photocoagulation and bevacizumab intravitreal injection combined with Krypton laser photocoagulation are all effective for DME. But treatment combined with Krypton laser photocoagulation could get significantly better postoperative visual acuity re-covery and significantly faster absorption of macular edema.

Objective To observe the mutifocal electroretinogram (mfERG) characteristics of rod and cone cells in patients with retinitis pigmentosa (RP) and to evaluate the function of photosensory cell.Methods The mfERG recording technique for rod cell in eight normal subjects (eight eyes) were established and the influence of different brightness lightstimulus in P1 wave amplitude were analyzed.The cone and rod cells mfERG of 38 eyes in 19 patients were recorded and then calculated positive ratio from local signal-noise ratio.The average visual acuity and P1 wave amplitude density of cone mtERG in different types were compared and statistically analyzed.Meanwhile,the changes in P1 wave amplitude of cone and rod mfERG in four quadrants also compared and analyzed.Results Rod cell mfERG in normal subjects can be recorded stably by using blue flashes with low light intensity as 0.04 cd/m~2.In patients with RP,the cone and rod cells mfERG can he detectd 65.790./00 and 10.51% respectively.P1 wave amplitude density in type I of cone cell mfERG was significantly higher than that in type Ⅱ (t=5.21,P=0.000).There were no differences in average visual acuity (t=1.15,P=0.612).P1 wave amplitude density in type Ⅰ was negatively related to IogMAR visual acuity (r=-0.48,P=0.04 ).The comparison of rod and cone cells mfERG in local wave characteristics showed that P_1 wave amplitude densities had spatial relationship in each area.Conclusions The results suggested highly variable central responses in cone cell in RP patients,higher positive recorded ratio in cone cell than rod cell and spatial correspondence between the function of reserved cone and rod cells.

Objective To investigate the relationship between electrophysiological and morphological properties of neurons in visual cortex of developing rat, speculate the coincided degree between electrophysiological and morphological change and realize the mechanism of normal visual development. Methods Whole cell patch-clamp recording and intracellular staining were used to acquire cellular microelectrode recording in visual cortex from Sprague-Dawley rats (4~28 days old). The histological process was made. Results The differences of electrical feature between pyramidal cells and non-pyramidal cells were significant. The morphological maturity degree is different in developing visual cortex. Conclusion The different function of pyramidal and non- pyramidal cells in local integrition is reflected by their electrical feature in the process of visual development. In critical period of visual development, the coincision degree of the electrophysiological and morphological change in visual cortex is larger than that in the subcortex constructure.

Purpose To investigate the effects of human vitreous fluid on proliferation of cultured human retinal pigment epithelial (RPE) cells and vascular endothelial cell lines(VEC304). Methods Human RPE cells and VEC304 were cultured and treated in different human vitreous conditioned medium (VCM) with or without serum, vitreous volume concentrations of VCM were 1∶8, 1∶4 and 1∶2. Cells proliferation was assayed by tetrazolium (MTT) colorimetry at the 2nd, 4th and 6th day respectively. Results In the presence of serum, 1∶4, 1∶2 VCM had a significantly stimulative effect on RPE cells proliferation compared with control group at the 2nd, 4th, and 6th day retrospectively ( P

Objective To describe cultured human retinal pigment epithelial (RPE) cells transdifferentiation and investigate the effects of human vitreous fluid on the morphologic and cytoskeleton changes of RPE cells in vitro. Methods Cytoskeleton characteristics in the 2 nd, 5 th, 8 th passage of RPE cells in normal culture, which included cytokeratin 18 (CK18) and ?-smooth muscle actin (?-SMA) were analyzed by Western blot. RPE cells were cultured in human vitreous-conditioned medium (VCM) at the concentration of 1∶4 for 6 days, morphologic changes were examined by light and electron microscopy, and cytoskeleton characteristics were analyzed by imunocytochemistry and Western blot. Results During culture in vitro, RPE cells lost epithelial characteristics and aquired fibroblast-like phenotype. The expression of CK18 was the highest at the 5 th passage, and it decreased in the following passage, but ?-SMA increased gradually. The morphologic transdifferentiation from epithelial to fibroblast-like cells of RPE was accelerated by VCM. Ultrastructural changes such as decreased microvilli and gradually increased rough endoplasmic reticulum and Golgi complex were found during the cultivation. CK18 produced by RPE cells decreased in VMC (P

Objective To observe safety of intravitreal injection of mouse nerve growth factor and its distribution in retina in rabbits .Methods The behavioral observation ,slit lamp examination ,fundus examination ,eye B ultrasonic and histopathological ex‐amination were carried out on 1 ,7 and 30 d after intravitreal injection 30 μg/100 μL mNGF to determine the safety in eye .The dis‐tribution and peak time in retina were investigated at 15 ,30 min ,1 ,3 ,6 ,8 ,12 ,24 ,48 h after intravitreal injection 125 I‐NGF 30 μg/100 μL .Results No abnormal changes were found in their cornea ,lens ,vitreous body and retina after mNGF intravitreal injection . And the each layer of retinal cells layout were regular according to the result of morphological observation on 30 days after treat‐ment .The peak concentration of mNGF in retina and the highest in whole eye was (118 .32 ± 18 .74)% ID/g and the peak time was at 3 hour after injection .Conclusion It is safe for intravitreal injection of mNGF and mNGF could gather in retina quickly after in‐travitreal injection .

Objective According to the basic theory of Yin and Yang,acute urticaria should be treated with divergent medicine instead of convergence medicine.In order to prove the theory,we carry out the quantifiable and repeatability traditional Chinese medicine screening tests in acute urticaria patients,with double controls of experimental group (divergent Chinese medicine),control group (convergence Chinese medicine)end instrumental quality-control standard substance MEBE.Methods Germany MORA-Super bio-resonance instrument (medical mode) as adopted as objective index.32 cases of acute phase urticaria were selected and treated with divergent or convergence Chinese traditional medicine,to observe the results whether conform to traditional Chinese medicine basic theory of Yin and Yang (That is whether the acute phase urticaria should be treated with divergent Chinese medicine).Results Through the 32 cases repeat experiments,22 out of 23 kinds of Divergent Chinese medicine have been screened out in experimental group; while none has been screened out in 12 kinds of convergence Chinese medicine in control group.There was significant difference between two groups (P＜0.01).The result was entirely consistent with TCM basic theory of opposite relationship between yin and yang.The top five divergent Chinese medicines were divaricate saposhnikovia root,cicada slough,catnip,shrub chastetree rruit,and lily magnolia.Conclusion Treated acute urticaria with divergent Chinese medicine conforms to the basic theory of traditional Chinese medicine.

Objective To isolate and cultivate Long-Evans rat bone mesenchymal stem cells (BMSCs) and identify their biological characteristics. Methods BMSCs were obtained from femurs of Long-Evan rats and dealt with 0.85 NH4Cl. The cell growth curve and cell cycle were measured and surface antigens were detected by flow cytometry. Osteoblast differentiation was studied by Alizarin red staining. Results BMSCs had active proliferative ability. The growth velocity of passage 5 cells was slower than that of passage 3 cells. The cell cycle analysis showed that 93.79 of BMSCs was at G0/G1 phase. BMSCs were positive for CD29 and CD44, but negative for CD45. Alizarin red staining of BMSCs after osteoblast induction was positive. Conclusion Highly purified BMSCs obtained from Long-Evan rats by 0.85 NH4Cl show stable biological properties.