Objective To investigate whether there is latency of herpes simplex virus 1 (HSV-1) in the ciliary body of normal human eyes. Methods Parts (1/3) of the 15 fresh eyeballs from normal healthy donors were used for immunohistochemical staining to exclude from acute infection of HSV-1. After the residual ciliary bodies were taken out, 1/3 was centrifugated for co-culture with Vero cells, and DNA was extracted from the other 2/3 for PCR and gel electrophoresis for the detection of HSV-1 DNA and latency associated transcript (LAT). Results All the 15 samples were negative of immunohistochemical staining and the results of co-culture of Vero cells with the supernatant containing the ciliary body tissues were also negative. No pathological changes in the cells were observed. In the 15 samples, no. 4, 5, 7 samples were positive of HSV-1 DNA but negative of HSV-1 LAT, whereas the other 12 samples were negative of HSV-1 DNA and HSV-1 LAT. Conclusion HSV-1 DNA exists in the normal human cilicary body.

Objective To investigate the expression of matrix metalloproteinases and tissue inhibitor of metalloproteinases in active Mooren's ulcer corneas.Methods An indirect immunofluorescent technique was used to visualize the presence of matrix metalloproteinase-1,2,3,9 and tissue inhibitor of metalloproteinase-1,2 in active Mooren's ulcer corneas from 6 patients and normal human corneas from 4 corpses.The results were further analyzed by an image-processing system.Results Only TIMP1,2 were detected in normal corneal epithelium while MMP-1,2,3,9,and TIMP-1,2 all were found in Mooren's ulcer corneas.MMP-2,TIMP-2 expressed in epithelium,MMP-3 expressed in stroma,MMP-1,9,and TIMP-1 expressed in both epithelium and stroma.Conclusion In active Mooren's ulcer cases,corneal lesion and corneal wound healing coexist,but the destructive reaction is greater than wound healing.Over-expression of MMP-9 and unbalanced regulation of MMPs/TIMPs may play an important role in the Mooren's ulcer of this stage.

Objective To compare the locations and levels of collagen type Ⅲ, laminin, and fibronectin in Mooren's ulcer and normal human corneas. Methods An indirect immunofluorescent technique and immunohistochemical staining were used to determine the distribution of collagen type Ⅲ, laminin, and fibronectin. The positive results were quantitatively analyzed with an image-processing system. Results Collagen type Ⅲ was not detected in the normal cornea. However, the staining could be seen in the corneal stroma close to Mooren's ulcer focus. Laminin was expressed faintly in the basement membrane of the normal cornea and the positive expression increased in the basement membrane and Bowman's membrane of Mooren's ulcer. In the normal cornea basement membrane, fibronectin was located continuously. In Mooren's ulcer basement membrane, however, fibronectin could not be found in all sections in which one expressed in epithelium and the others in stoma close to the ulcer focus. Conclusion Epithelial basement membrane may play a potential role in the pathogenesis of Mooren's ulcer.

0.05).Conclusion Vitrectomy significantly alters the stability of tear film.The tear film function returned to preoperative conditions in 3 months after operation.

Objective To investigate the antimicrobial properties of human amniotic homogenate supernatant (HAHS) in order to found a theoretical base for the use of human amniotic membrane in ophthalmological field. MethodsFresh human amniotic membranes were used to make HAHS and acellular amniotic membranes. Then, we observed their antimicrobial effects and antimicrobial spectrums, compared the antimicrobial capacity with 10 commonly used antibiotics in eyes, and investigated the effects of time, temperature and pH value on the antimicrobial capacity. Finally, transmission electron microscopy (TEM) was used to explore the possible targeting site of the antibiosis. ResultsHuman amnion membrane contained antimicrobial components locating in its epithelial cells. HAHS had a broad antimicrobial spectrum and was steady in nature. Its antimicrobial capacity was stronger than those of sulfasulfonamide, chloromycetin and cefuroxime sodium. TEM indicated that the antimicrobial effect were exerted through plasma membrane of microorganism. ConclusionHAHS can be an effective and convenient treatment for ocular surface infectious diseases. Traditional amnion transplantation should employ fresh human amniotic membranes containing complete epithelial lamina to reconstruct the ocular surface.

Objective To explore the possibility of cytochrome P-450 3A5 (CYP3A5) genotype as a major factor to guide individualized employment of cyclosporin A(CsA) through a comparative study of CsA concentrations in the peripheral blood of hemopoietic stem cell transplant recipients with different CYP3A5 genotypes. Methods Olymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to genotype CYP3A5 gene, and CsA concentrations were detected by a commercial fluorescence polarization immunoassay. Result There were significant differences in CsA concentrations, including standardized trough concentrations C0 and two h peak concentrations C2 , in the two CYP3A5 genotypes found in our samples, and both C0 and C2 in wild homozygotes were lower than heterozygotes. Conclusion CYP3A5 polymorphism is highly associated with CsA concentrations in hemopoietic stem cell transplant recipients, and CYP3A5 genotype may be a predictor to the dose requirement before clinically employment of CsA.

Objective To evaluate the murine model of dry eye induced by hyperosmolar saline. Methods Sixty female BALB/c mice at the age of 6 -8 weeks were randomly divided into blank group,control group and experimental group,20 in each group. Mice in control and experimental groups were treated with 308 mOsmol/L and 500 mOsmol/L sodium chloride solution,respectively,5 times a day. Mice in blank group were not treated with sodium chloride solution. Schirmer test,fluorescein staining,corneal scoring,rose bengal staining,tear ferns experiment,corneal epithelial HE staining and thickness measurement,conjunctival epithelial PAS staining and Goblet cell counting were conducted on days 0,7,14,28,and 42,respectively. Corneal surface was observed by scanning electron microscopy on day 42. Results No significant difference was found in the above parameters on day 0 between the two groups. On day 7,the volume of tears was significantly smaller in experimental group ( 2. 3 ? 0. 4 mm) than in blank group ( 3. 0 ? 0. 5mm) and control group ( 3. 1 ?0. 5 mm) ( P

BACKGROUND: There are various reports on studies of tear secretion and tear film function in patients with diabetes mellitus over the past. In recent years, tear proteins have drawn more and more attentions on evaluation of tear film function.OBJECTIVE: To observe the contents of main tear proteins and basal tear secretion of patients with type 2 diabetes mellitus and normal persons so as to probe into tear secretion and tear film function of patients with diabetes mellitus.DESIGN: Case-control observation was designed.SETTING: Department of Ophthalmology, Southwest Hospital, Third Military Medical University of Chinese PLA.PARTICIPANTS: Totally 50 cases (100 eyes) of type 2 diabetes mellitus were employed, which were diagnosed in Department of Ophthalmology and Department of Endocrinology, Southwest Hospital of Third Military Medical University of Chinese PLA from December 2001 to December 2002. They were free from ocular surgical and laser treatment, local medication in recent period and contact lens. Of those, there were 25 cases (50 eyes) in proliferating diabetic retinopathy group and 25 cases (50 eyes) in nonproliferating diabetic retinopathy group. In addition, 25 cases (50 eyes) of normal persons with matched age and sex were taken as the control group.There was no significant difference in age and sex among 3 groups (x2=0.024,0.321 ;P ＞ 0.05). All of the participants were in the know before the experiment.METHODS: [1] Tear collection: 10 cases (20 eyes) were randomized from two diabetic groups and the control successively. Capillary pipette method was used to collect non-irritative tear 10 μL from lower lacrimal punctum that was preserved in refrigerator at -20 ℃ (＜ 1 month). [2] Determination of total tear protein amount: Lorry method was used to determine the concentration of total tear protein, in which, calf serum albumin was taken as the criteria. [3] Determination of contents of main tear proteins: SDS-PAGE (sod.dodecyl sulfate-polyacrylamide gel electrophoresis) was used and Coomassie brilliant blue staining and Bio-Rad imaging analyzing system were applied for the analysis of isolated protein strips in quality and quantity. [4] Determination of rupture time of tear film: glass rod was used to get 20 g/L fluorescein sodium and drop in conjunctival sac. The examined person was required to blink gently for several times and open the eyes naturally and stare forward. That the first ruptured "black hole" was discovered on the complete tear film was taken as the rupture time of tear film. [5] Experiment of basal tear secretion: No.41 Whatmann filtering paper was used, folded in 5 mm, and placed at 1/3 of conjunctiva in the lower eyelid. Five minutes later, the paper was removed and length of it after wetting was measured. [6]Experiment with rose Bengal staining: glass rod was used to get 10 g/L rose Bengal and drop in conjunctival sac. After eyes blinking for several minutes, the observation was performed with green light filter under slit lamp (evaluation criteria: corneal conjunctiva of palpebral fissure stained+, stained to the inferior bulbar conjunctiva++,stained to the superior bulbar conjunctiva). Dry eye disease was diagnosed indirectly with red-stained epithelial cells and mucin.MAIN OUTCOME MEASURES: [1] Concentration of total tear protein. [2]Concentrations of various main tear proteins. [3] Rupture time of tear film. [4]Value of basal tear secretion. [5] Positive rate of rose Bengal staining.RESULTS: Totally 50 cases (100 eyes) in diabetes groups and 25 cases(50 eyes) in the control group all entered result analysis. [1] Concentration of total tear protein: there was no significant difference among 3 groups (P＞ 0.05). [2] Concentrations of various main tear proteins: the results of lysozyme, lactoferritin and tear specific prealbumin in proliferating diabetic retinopathy group were lower remarkably compared with the control[ (0.94±0.21)vs ( 1.33±0.31 )g/L , ( 1.10±0.24)vs ( 1.67±0.43 )g/L, (0.98±0.22) vs (1.49±0.32)g/L, P ＜ 0.01]. Compared with non-proliferating diabetic retinopathy group, the results of lactoferritin and tear specific prealbumin were lower (P ＜ 0.05). There was no significant difference in human serum albumin among 3 groups (P ＞ 0.05). [3] Rupture time of tear film:compared with non-proliferating diabetic retinopathy group and the control, the rupture time of tear film in proliferating diabetic retinopathy group was reduced significantly [(7.68±2.21)s vs (9.92±2.37)s and(10.80±2.23)s,(P ＜ 0.01 )]. [4] Value of basal tear secretion: the value in proliferating diabetic retinopathy group was less significantly than that in non-proliferating diabetic retinopathy group and the control [ (8.00±2.10)vs( 11.02± 1.97 )mm and ( 12.17±2.08 )mm, P ＜ 0.05]. [5] Positive rate of rose Bengal staining: the positive rate in proliferating.diabetic retinopathy group was higher significantly than non-proliferating diabetic retinopathy group and the control (48% vs 24% and 14%, P ＜ 0.05, P ＜ 0.01).CONCLUSION: It is suggested in the results of this paper that abnormal tear secretion and tear film function are apt to present in patients with type 2 diabetes mellitus and the declined tear film function is more remarkably in the patients with proliferating diabetic retinopathy specially. SDS-PAGE benefits the discovery of changes in tear proteins in diabetic patients.

One strain of Hantavirus(HV) was isolated from patients with hemorrhagic fever with renal syndrome (HFRS) in Zhejiang province and its S-segment was cloned and submitted to nucleotide sequence analysis in order to determine the type of strain and extent of genetic variation for further study on its evolution and mutation .The HV antigen was detected from mouse lungs in endemic area by direct immunofluorescene test and the HV-positive sample of mouse lung was inoculated to Vero-E6 cells to isolate virus. The total cellular RNA was extracted from infected cell culture and the S segment gene was amplicated by RT-PCR. Then, the purified PCR product was cloned into T vector for sequencing. The result showed that the full-length sequence of the S segment was 1 700 bp with one open reading frame (ORF) encoding 429 amino acids. Comparison with Hantavirus HTN type showed 85.7%-91.9% homology at the nucleotide level. In comparison with the SEO type of viruses, the homology at nucleotide level was shown to be 71.2%-75%. This new HV strain may be typed as to HTN virus and may be a new subtype of this virus.

Objective To investigate the effects of chondroitin sulfate proteoglycans (CSPGs) degradation induced by chondroitinase ABC (ChABC) on the rat flash visual evoked potential (F-VEP) around the end of the critical period of visual development.Methods A total of 30 Long Evans rats were randomly divided into 3 groups according to their age of 14,21 and 35 d.Then in each group,5 animals served as CSPGs degradation model induced by ChABC microinjection into 5 sites of the rat cortex [750 nL (48 U/ml) for 1 site,for 2 times at a 3-day interval],and the other 5 rats served as normal control and received normal saline microinjection.Immunofluorescence microscopy and laser scanning confocal microscopy were used to determine the establishment of animal model.F-VEPs were evaluated in 7 d after first microinjection.Results In the normal rats,the latency of F-VEP main wave became shorter with the increase of their age,the number was [(63.5?10.1),(50.8?6.4),(44.9?6.3) ms,P