To study the secretion of epidermic growth factor (EGF) in tears from patients with herpes simplex virus keratitis (HSVK), radio-immunoprecipitation was used to detect the value of EGF in human reflex tears from 40 cases as control group (group A), 49 cases of acute HSVK (group B) and 31 cases of convalescence HSVK (group C). Results showed that the value of EGF in reflex tears from the patients in the group B was obviously increased as compared with that in the group A and group C (P<0.001). The value of EGF in tears in the group C was higher than in group A (P<0.05). It was concluded that the value of EGF in reflex tears from HSVK patients was higher than that from normal people.

With the development of biological techniques,the study on the pathogenesis of disease-causing genes of congenital cataracts has substantial progress.Some positive results of screen of mutation gene in congenital cataract family has been reported,but the report of negative result is rate.ObjectiveThe present study attempts to screen the mutation of CRYAA,CRYAB,CRYA1/A3,CRYBB2,CRYGC and CRYGD gene in a Chinese family with autosomal dominant congenital cataract. MethodsThe periphery blood samples were exacted from 8 patients of 4 generations of with congenital cataract in this family.The complete coding region and intron spliced sites of CRYAA,CRYAB,CRYA1/A3,CRYBB2,CRYGC and CRYGD were amplified with polymerase chain reaction (PCR),and the products of PCR were directly sequenced.The control blood samples were from 10 normal subjects.This study followed the Declaration of Helsinki.Written informed consent was obtained from all of the patients.ResultsThe patients were found in each generation in this family and the mode of inheritance was in accordance with the characteristic of autosomal dominant inheritance.The sequence of amplified genetic fragments of CRYAA,CRYAB,CRYA1/A3,CRYBB2,CRYGC and CRYGD genes were inaccordance with those of normal subjects and GeneBank.No any mutation loci was found in all of the patients of this family.ConclusionCRYAA,CRYAB,CRYA1/A3,CRYBB2,CRYGC and CRYGD genes is not the causing-disease genes in this family.

Objective To analyze and compare the characteristics and differences of corneal endothelial cells of rhesus monkey and tree shrew eyes.Methods Corneal endothelial cells of 6 healthy rhesus monkeys (12 eyes) and 20 healthy tree shrews (40 eyes) were measured using a non-contact SP3000P specular microscope.Eight parameters were de-termined and compared with relevant parameters of human eyes reported in the literature, including minimum cell area (Smin), maximum cell area (Smax), average cell area (Savg), standard deviation of cell area (SD), coefficient of variabili-ty ( CV) , cell density ( CD) , hexagonality percentage ( HG%) and central corneal thickness ( CCT) .Results The ima-ging and measurement of all parameters could be completed in a short time both in rhesus monkeys and tree shrews.The time spent in the two kinds of animals was not significantly different.The CCT was ( 449.2 ±12.8 ) μm and ( 262.4 ± 24.6) μm, Smin was (120.4 ±26.3) S/μm2 and (153.2 ±42.9) S/μm2 , Smax was (705.0 ±130.8) S/μm2 and (468.7 ±109.3) S/μm2 , Savg was (351.1 ±26.1) and (295.4 ±18.9) S/μm2 , SSD was (113.1 ±27.4) and (75.9 ±27.3) S/μm2, CV was (31.9 ±6.0) and (25.3 ±8.3), CD was (2874.2 ±203.8) p/cell· mm-2 and (3399.3 ±224.7) p/cell· mm-2 , and the HG% was (58.6 ±9.1) and (94.0 ±9.7) in the rhesus monkeys andt tree shrews, respectively. The differences of all the above parameters between rhesus monkeys and tree shrews were statistically significant ( P<0.05 for all) .The cornea of tree shrews was significantly thinner than that of rhesus monkeys.The area and coefficient of varia-bility of tree shrews were smaller to those of rhesus monkeys, while the cell density and hexagonality percentage were higher than those of rhesus monkeys.Compared with human eyes, the CCT, CV and HG%in rhesus monkeys were highly simi-lar, while the CD was lower than that of human eyes.The CCT in tree shrew was only 60%of the rhesus monkey eyes and 50%of human eyes, while the CD and Savg were similar to that of human eyes in the 10-20 years old group.Conclu-sions The morphology and parameters of corneal endothelial cells in rhesus monkeys and tree shrews are significantly dif-ferent.There are similarities and differences among the human, rhesus monkey and tree shrew corneal endothelial cells. Both rhesus monkeys and tree shrews are appropriate experimental animals feasible for researches on human corneal endo-thelial diseases.

OBJECTIVE:To systematically review the efficacy and safety of Xiyanping injection versus Ribavirin injection in the treatment of herpangina in children,and provide evidence-based reference for clinical treatment. METHODS:Retrieved from PubMed,Cochrane Library,EMBase,VIP Database,CJFD,Wanfang Database and CBM,randomized controlled trials (RCT) about efficacy and safety of Xiyanping injection versus Ribavirin injection in the treatment of herpangina in children were collected. Meta-analysis was performed by using Rev Man 5.3 software after data extracting and quality evaluating by modified Jadad. RE-SULTS:Totally 14 RCTs were enrolled,involving 1 939 patients. Results of Meta-analysis showed total effective rate [OR=4.69, 95%CI(3.36,6.55),P<0.001],fever clearance time [MD=-1.36,95% CI(-1.60,-1.12),P<0.001],herpes regression time [MD=-1.34,95%CI(-1.61,-1.06),P<0.001],hospitalization time [MD=-1.88,95% CI(-3.68,-0.07),P=0.04] and sali-vation disappearance time [MD=-1.07,95% CI(-1.30,-0.84),P<0.001] of Xiyanping injection were significantly better than Ribavirin injection,there were statistically significant differences. And there was no significant difference in the incidence of ad-verse reactions [OR=0.56,95% CI(0.31,1.03),P=0.06]. CONCLUSIONS:The efficacy of Xiyanping injection is better than Rib-avirin injection in the treatment of herpangina in children,with similar safety.

Objective To observe the clinical effect oflamellar keratectomy+conjunctival flap+implanted artificial eyein stimulating the orbital and conjunctival sac growth. Methods A retrospective case study: 12 cases (12 eyes) with congenital microphthalmos in the Fourth Affiliated Hospital of Kunming Medical University in 2009-2013 were selected. In these cases, there were 11 cases of microphthalmos, and 1 patient due to congenital absence of the eye without surgery, were given direct implant of the artificial eye;7 patients without significant stenosis in conjunctival sac,received thelamellar keratectomy+conjunctival Flap+implantation of artificial eye, 4 patients with conjunctival sac stenosis recieved thelamellar keratectomy+conjunctival flap+implanted artificial eye+eyelid suture. Results For stunted children who couldn't wear a prosthetic eye, after treated withlamellar keratectomy + conjunctival flap + artificial eye implantation, the conjunctival sac developed well, cornea was covered with conjunctiva well and no exposure,the appearance and volume of orbit was also improved. ConclusionLamellar keratectomy+conjunctival flap+artificial eye implantsurgery is an effective way to promote orbital and conjunctival sac development of the children with congenital microphthalmos.

In order to investigate the effect of nerve growth factor (NGF) on the proliferation of rabbit corneal endothelial cells and epithelial cells, the in vitro cultured rabbit corneal endothelial cells and epithelial cells were treated with different concentrations of NGF. MTT assay was used to examine the clonal growth and proliferation of the cells by determining the absorbency values at 570 nm. The results showed that NGF with three concentrations ranging from 5 U/mL to 500 U/mL enhanced the proliferation of rabbit corneal endothelial cells in a concentration-dependent manner. 50 U/mL and 500 U/mL NGF got more increase of proliferation than that of 5 U/mL NGF did. Meanwhile, 50 U/mL and 500 U/mL NGF could promote the proliferation of the rabbit corneal epithelial cells significantly in a concentration-dependent manner. However, 5 U/mL NGF did not enhance the proliferation of epithelial cells. It was suggested that exogenous NGF can stimulate the proliferation of both rabbit corneal endothelial and epithelial cells, but the extent of modulation is different.
Cell Proliferation/*drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelium, Corneal/*cytology ; Epithelium, Corneal/*cytology ; Nerve Growth Factor/*pharmacology

Objective To establish a tree shrew model of Fusarium solani keractitis by injecting Fusarium solani conidia into the corneal stroma.Methods Fusarium solani was inoculated into Sabouraud culture medium and incubated at 26℃ for 7 days.Fungal suspension was collected and the number of spores was adjusted to 1 × 1010 CFU /mL on the blood cell count plate.Forty healthy tree shrews were randomly divided into experimental group (n=30) and control group (n=10).In the experimental group, 50 μL of fungal spore suspension was injected into the cornea center with a 29G needle, and 50 μL saline was injected in the control group.The models were evaluated by anterior segment photography, in vivo confocal microscopy, histopathology, and corneal tissue culture.Results The fungal infiltration, the degree of edema of corneal epithelial and endothelial cells, and the number of mycelium were positively correlated with time.The number of infiltrating inflammatory cells, mainly, neutrophils, reached a peak on the 7th day after modeling.The mycelial growth was parallel to the stromal fibers.After the successful establishment of the model, the corneal tissue culture showed the growth of Fusarium solani.The successful rate of modeling was 86%.Conclusions The tree shrew model of Fusarium solani keratitis is established by injecting spores of Fusarium solani into the cornea.

Background Elevated intraocular pressure leads to the loss of retinal ganglion cells and vigorous reaction of retinal glial cells.The expression of nestin in retinal glial cells secondary to hypertention and its significance are unclear.ObjectiveThis study aim to investigate the expression of nestin in retinal glial cells (RGCs) in ocular hypertention rats.Methods The ocular hypertention models were established by cauterizing the limbus-draining veins in the right eyes of 42 SD rats,and a conjunctival incision in the left eyes of the rats served as the sham group.The intraocular pressure (IOP) was measured with the Tono-Pen XL tonometer.The number of RGCs in the rats with ocular hypertention was counted.The expression of the nestin protein in RGCs was semi-quantitatively analyzed using Western by immunochemistry.Double immunofluorescence was carried out to evaluate the the confocal laser scaning microscope.Results Significant differences were found in the IOP between the model group and the sham group at various time points (P＜0.05).In 1 week to 3 weeks after operation,the number of RGCs significantly declined in the model group compared with the sham group (P＜0.05).Immunochemistry showed that from 2 hours through 1 week after operation,the expression of nestin was gradually enhanced in the model group in comparison with the sham group.Western blot revealed that the expression of the nestin protein reflected a similar tendency to that of immunofluorescence.The increased introcular pressure as manifested by the induced expression of nestin.Immunoelectron microscopy also confirmed the induced expression of nestin especially at their end-feet suggests a potential neuroprotective mechanism in neuronal degeneration.Nestin may be a useful biomarker for retinal injury study.

Objective To observe the changes of Th1 / Th2 inflammatory factors in the aqueous humor of tree shrews with fusarium solani keratitis,as well as to explore the relationship between Thl / Th2 inflammatory factors and inflammatory response in fusarium solani keratitis.Methods Forty healthy tree shrews were randomly divided into experimental group(n =30) and control group (n =10).Fusarium solani was inoculated into sabina culture medium and cultured at 26 ℃ for 7 days,and then the fungal suspension was collected and the density of spores was adjusted to 10 × 109 CFU · mL-1.In the experimental group,50 μL fungal spore suspension was injected into the center of the cornea stroma,while the control group received the same amount of saline.Next the levels of cytokines including interleukin (IL)-1 β,IL-6,IL-4 and IL-10 were analyzed by flow cytometry on day 3,day 7,day 14 after successful modeling,and the changes in types of infiltrating cells were observed by histopathological examination.Results The expression level of IL-1 β and IL-6 (Th1 type cytokines) was the highest on day 7,and the difference was statistically significant at each time point when compared with the control group (all P ＜ 0.05).The expression level of IL-10 (Th2 type cytokines) was the highest on day 14,and the difference was statistically significant at each time point when compared with the control group (all P ＜ 0.05).The difference in IL-4 expression was statistically significant on day 7 (P ＜ 0.05).In addition,histopathological examination showed that the number of infiltration cell reached its peak on day 7,mainly neutrophils,and fungal hyphae were observed to be parallel to the matrix fibers at each time point.Conclusion The proinflammatory cytokines IL-1β and IL-6 and the anti-inflammatory cytokines IL-10 may play an important role in the molecular mechanism of inflammatory response of fusarium solani keratitis in tree shrews.

Objective To study the clinicopathological significance of the expression of glutameta decarboxylase 65(GDA65) and protein kinase C(PKC) in the central cancer tissues, cancer edge tissues, paracancerous liver tissue and non-cancer liver tissues. Methods The expression of GDA65 and PKC were detected by immunohistochemical method in 10% neutral formalin- fixed and routinely paraffin-embedded sections in 37 hepatic cancer specimen. Results The positive rate and the score of GDA65 and PKC in the cancer tissues were significantly higher than that in the paracancer tissues or non-cancer liver tissues, but the PKC expression was no difference between the central cancer tissues and the cancer edge tissues . The expression of GDA 65 was related to the pathological types, differentiated degrees, liver cirrhosis or metastasis of hepatocarcinomas. No correlation was found between the expression of PKC and the clinicopathological features of hepatocarcinomas. Conclusions The expression of GDA65 and PKC might be closely related to the carcinogenesis of hepatocarcinoma, they might be important biological markers of hepatocarcinoma.