The effect of hydroxyprolpyl methylcelulose (HPMC) injected into anterior chamber on filtering angle tissues was studied. Twenty-four rabbits were randomly divided into two groups: the experimental group included 40 eyes receiving HPMC which was injected into anterior chamber and the control group included 8 eyes (one served as normal control and the remaining were injected with balanced salt solution). According to the intaocular pressure (IOP) was normal or elevated ,the experimental group was further divided into group A and B. The tissul specimen of filtering angle were collected to perform pathological examination at 5 days, 3 weeks ,and 10 weeks after injection.During the early period, in groupA,the trabecular meshwork dilated slightly, the vacuoles increased in endothelial cells of inner wall of Schlemm's canal. In group B, the space of trabeculum broadened, there were accumulation of collagen and intercellular fibrosis, the vacuoles significantly increased in endothelial cells of inner wall of Schlemm's canal. But the change mentioned above all recovered to normal appearance on the later stage of long term following observationHPMC injected into anterior chamber may cause temporary elevation of IOP and pathological change of filtering angle, but the change was reversible. HPMC is not toxic or destructive to the tissues of anterior chamb angle.

This study investigated the inhibitory effect of grape seed proanthocyanidin extract (GSPE) on selenite-induced cataract formation in rats and the possible mechanism. Eighty 8-day-old Sprague-Dawley rats were divided randomly into 5 groups: control group, model group, three GSPE groups (low dose, medium dose and high dose). Control group received subcutaneous injection of physiological saline. Model group was given subcutaneous injection of sodium selenite (20 μmol/kg body weight) on the postpartum day 10, and once every other day for consecutive three times thereafter. GSPE treated groups were respectively administered GSPE at doses of 50, 100, and 200 mg/kg body weight intragastrically 2 days prior to the selenite injection (that was, on the postpartum day 8), and once daily for fourteen consecutive days thereafter. The opacity of lenses was observed, graded and photographed under the slit lamp microscopy and the maximal diameter of the nuclear cataract plaques was measured. The lenses were analyzed for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), malondialdehyde (MDA), calcium (Ca(2+)), nitric oxide (NO) and anti-hydroxyl radical ability (anti-OH(-)). The histomorphology of lenses was observed with HE staining under a light microscope. The levels of calpainII, and iNOS protein and mRNA expression in lenses were detected by using immunohistochemistry and real-time quantitative RT-PCR. The results showed subcutaneous injection of sodium selenite led to severe nuclear cataract in model group, and the achievement ratio of model group was 100%. As compared with model group, the degree of lenses opacity and the maximal diameter of nuclear cataract plaques were significantly reduced in GSPE-treated groups. Moreover, we observed selenite treatment caused a significant decrease in the activities of antioxidative enzymes (SOD, CAT, GSH-PX) and anti-OH(-) ability, accompanied by a significant increase in the levels of MDA, NO, Ca(2+) as well as iNOS, and calpainII protein and mRNA expression. Administration of GSPE could dose-dependently preserve the activities of these antioxidative enzymes and anti-OH(-) ability, accompanied by a significant reduction in the levels of MDA, NO, Ca(2+) as well as iNOS, and calpainII protein and mRNA expression. These results suggested that GSPE markedly prevented selenite-induced cataract formation probably by suppressing the generation of lipid peroxidation and free radicals as well as the activation of iNOS, and calpainII in the lenses.

To investigate the neuroprotective effect of melatonin (MT) on retinal ganglion cells (RGCs) in rats with ischemia reperfusion injury (RIR), 24 healthy SD rats were randomly divided into two groups: group A and group B. RIR model was induced in the left eyes by increasing the pressure of the anterior chamber. Group A was treated with 10 % alcohol- normal saline (1 mL/kg/d, ip), while group B was treated with 0.5 % MT (1 mL/kg/d, ip). On the basis of the time interval between the left eyes RIR and the sacrifice, rats in both group A and group B were further divided into 3 subgroups: groups A1 and B1 (days 7), groups A2 and B2 (days 14), groups A3 and B3 (days 30), with 4 rats in each subgroup. 7 day before the sacrifice, 3 % fluorogold was bilaterally injected into superior colliculi and geniculate body. The eyes were enucleated after being sacrificed, and mounting of the retina from both eyes was performed on a slide and observed under a fluorescence microscope. Four photos were taken from each of the four quadrants of the retina. The labeled-RGCs were counted by using a computerized image analyzer. The rate of the labeled-RGCs was used for statistical analysis. Our results showed that, in group A, the rate of the labeled-RGCs was (77.16 +/- 6.35) %, (65.53 +/- 7.01) %, (53.85 +/- 4.38) % on day 7, 14 and 30. In group B, the rate of the labeled-RGCs was (81. 33 +/- 9.27) %, (79.80 +/- 8.36) %, (80. 34 +/- 11.05) % on day 7, 14 and 30. In group B, which was treated with MT after RIR, the rate of labeled-RGCs was significantly higher than that of group A on day 14 and day 30 (P<0.05). It is concluded that, in the RIR rats, MT therapy could increase the survival rate of the RGCs and could rescue and restore the injured RGCs.

Objective This study was to observe the effects of connective tissue growth factor(CTGF) on migration and transdifferentiation of bovine lens epithelial cells(BLECs). MethodsThe culture and identification of BLECs adopted the method of Hu(reference 1).The 2-3 passages of BLECs were collected and used in this experiment at the concentration of 1×10~6 cells/hole.The free-serum DMEM containing 0.1 ng/L,0.5 ng/L,1.0 ng/L of CTGF was added into medium for 24 hours in different experimental group respectively,and only equal volume of free-serum DMEM was added in control group.Expression of α-smooth muscle actin(α-SMA) mRNA and protein in the BLECs were examined by semiquantitative RT-PCR and Western blot,respectively.The transwell inserts were used to evaluate the migration ability of BLECs. ResultsThe expression of α-SMA mRNA in cultrued BLECs was gradually increased in different concentrations of CTGF groups.Compared with control group,the expression of α-SMA mRNA in experimental group was significantly enhanced (F=66.56,P＜0.01).The expression of α-SMA protein followed the same pattern(F=65.43,P＜0.01).The migration ability of BLECs was obviously elevated after CTGF stimulation under the light microscope.The migration rate of BLECs was considerably increased in experimental group compared with blank control group (t=51.7,P＜0.01).ConclusionCTGF promotes the migration and transdifferentiation of BLECs at a dose-dependent manner in vitro.CTGF plays an important role in the formation of posterior capsule opacification.

BackgroundOur previous research and other reports disclosed that the expression of tissue transglutaminase(tTG)in lens epithelial cells(LECs) of patients with cataract is enhanced,indicating tTG is related to formation of posterior capsule opacification(PCO).ObjectivePresent study is to observe the effect of tTG specific inhibitor monodansyl-cadaverineon(MDC) on the expression of fibronectin(FN) and collagen Ⅳ(Col-Ⅳ) induced by TGF-β_2 in human LECs.MethodsHLE-B3 was cultured in vitro in DMEM containing 10% fetal bovine serum and then were divided into 5 groups.The free-serum culture was used as normal control group.Free-serum culture containing 10μg/L TGF-β_2 was utilized as treatment group.10μg/L TGF-β_2 plused 100μmol/L,200μmol/L and 400μmol/L MDC respectively in different concentrations as MDC-treatment group.Semiquantitative RT-PCR was used to assay the expression of tTG,FN and Col-Ⅳ in HLE-B3.A(tTG/β-actin),A(FN/β-actin) and A(Col-Ⅳ/β-actin) was calculated separately as the detecting indexes.ResultstTG,FN and Col-Ⅳ were positively expressed in cultured HLE-B3.The expression levels of tTG,FN and Col-Ⅳ in HLE-B3 were remarkably increased in the group with 10μg/L TGF-β_2 compared with normal control group(t=33.95,P<0.01;t=38.24,P<0.01;t=13.48,P<0.01).The expression levels of FN and Col-Ⅳ were gradually declined in 100,200 and 400μmol/L MDC groups in comparison with TGF-β_2 treatment(P<0.01).The significant differences were also found in the expressions of FN and Col-Ⅳ in HLE-B3 among 100,200 and 400μmol/L MDC groups(P<0.01).ConclusionMDC inhibits the expression of FN and Col-Ⅳ induced by TGF-β_2 in human LECs at a concentration-dependent manner.tTG may be involved in the formation of posterior capsule opacification through up-regulating the expressions of FN and Col-Ⅳ in human LECs.

Plasma HDL, HDL2 and HDL3 were determined in 95 untreated and 87 treated patients with NIDDM and 86 cases with IGT using Eder's modified method.The result showed that HDL, HDL2 and HDL3 were all reduced in these three groups in comparison with control group, but the average concentrations of all 3 subfractions of HDL showed no significant difference among these three groups. It also indicated that HDL abnormality may appear quite early in cases with abnormal glucose tolerance, such as in patients with IGT.

Flow cytometry analysis of DNA ploidy level and S-phase fraction (SPF) was carried out in 24 snap-forzen tissue specimens of ovarian carcinoma. The relationship of DNA ploidy level and SP-F with age,clinical stage, ascites, histologic type,pathological grade,lymphocyte infiltration and psammoma bodies was analyzed.Results showed that the relationship of DNA ploidy level and SPF with ascites and pathological grade was significant. Aneuploid portion and SPF in patients with ascites or pathological grade Ⅲ or Ⅱ were higher than those in patients with no ascites or pathological grade I. The author considers that DNA ploidy level and SPF of ovarian carcinoma can serve as a relatively independent objective index which reflects tumor's biological behaviours,and will be of great auxiliary value to early diagnosis and therapy of ovarian carcinoma.

To study whether cultured bovine trabecluar meshwork cells (BTMC) are capable of expressing tTG in protein and at mRNA level, BTMC were cultured in vitro and passaged three times, then the cells were transferred onto or cultured on sterile cover or submitted to isolation of RNA with Trizol, and the expression of tTG was detected by immunohistochemical technique and reverse transcription polymerase chain reaction (RT-PCR) respectively. Our results showed that tTG immunostaining was positive in the cytoplasm and rarely in the nucleus of cultured BTMC. No immunostaining was seen in the negative control. Moreover, a single RT-PCR amplified product whose sequence and size were in accordance with our known parameters was obtained. The expression of tTG in cultured BTMC was confirmed in protein and at mRNA level. BMTC is available more readily for the investigation of the relationship between tTG and primary open-angle glaucoma.
Cells, Cultured ; Glaucoma, Open-Angle/metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Trabecular Meshwork/*metabolism ; Transglutaminases/*biosynthesis ; Transglutaminases/genetics

To investigate the effect of proteasome inhibitor MG132 on the apoptosis of bovine lens epithelial cells (BLECs), the cells were treated with MG132 at different concentrations for12, 24 and 36 h. The cell viability was analyzed by MTT assay and the effect of MG132 on the apoptosis of BLECs was assessed by flow cytometry (FCM). The results showed that after treatment for the same period, the inhibitory effect of MG132 on BLECs proliferation was enhanced with the increment of the concentration of MG132 (0, 2, 5, 10, mumol/L) (P<0.05). The 50% inhibiting concentration (IC(50)) was 2.03 mumol/L when the BLECs were treated with MG132 for 36 h. MG132 also induced the apoptosis of BLECs obviously. FCM showed that the apoptosis index of the cells treated by MG132 at 2 mumol/L for 12 h was (20.24+/-1.51)%, and that of the control was (0.98+/-0.20)% respectively (P<0.01, n=3). It was concluded that MG132 could lead to apoptosis of BLECs. The decrease of proteasome activity may play an important role in the formation and development of cataract.

To study the effect of tTG fully phosphorothioated antisense oligodeoxynucleotides (tTG-ASDON) on tTG expression in cultured bovine trabecular meshwork cells (BTMCs) in vitro and explore a new treatment alternative for primary open angle glaucoma (POAG), the ASDON1 and ASDON2 complementary to the protein codogram region of tTG were designed, synthesized and phosphorothioated according to the secondary structure of tTG. The ASDON1 and ASDON2 were embedded in Lipofectamine and transfected into BTMCs. The untreated group served as negative controls. The expression of tTG in the mRNA and protein level were measured by semi-quantitative RT-PCR and immunohistochemical technique-Supervision method respectively. Our results showed that both the mRNA and the protein of tTG with tTG-ASDON and tTG-ASDON2 were significantly decreased as compared with that of the controls (P < 0.05). On the other hand, no significant difference was found between the ASDON1 group and the ASDON2 group. It is concluded that the expression of tTG mRNA and protein in cultured BTMC are down-regulated by tTG- ASDON. As a result, tTG-ASDON may be used for the treatment of POAG through the inhibitory effect on the expression of tTG.
Cells, Cultured ; Glaucoma, Open-Angle/metabolism ; Oligonucleotides, Antisense/*pharmacology ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Trabecular Meshwork/cytology ; Trabecular Meshwork/*metabolism ; Transglutaminases/*biosynthesis ; Transglutaminases/genetics ; Transglutaminases/*pharmacology