Objective To investigate the mechanism by which INKA2-AS1 promotes hepatocellular carcinoma(HCC)migration,invasion,and macrophage-related immunosuppression.Methods Based on the Cancer Genome Atlas Program(TCGA)-HCC data and Genotype-Tissue Expression(GTEx)databases,differentially expressed genes were analyzed in 374 HCC tissues and 160 normal liver tissues.Survival analysis was used to investigate the relationship between INKA2-AS1 and the prognosis of HCC patients.Cox regression analysis was performed on INKA2-AS1.The relationship between INKA2-AS1 and immune infiltration in HCC tissues was analyzed using the Tumor Immune Estimation Resource Database.A total of 90 HCC patients were recruited from the Department of Hepatobiliary Surgery,Chongqing Hygeia Hospital from January 2019 to January 2021.The HCC tissues and adjacent normal tissues were collected.Hepa 1-6 liver cancer cells were grouped into:normal control(NC),INKA2-AS1,siNC and siINKA2-AS1.INKA2-AS1 was overexpressed and silenced by transfection.The proliferation,migration and invasion of cells in each group were detected by CCK-8,EdU,TUNEL,wound healing and Transwell assays.Each group of Hepa 1-6 cells were co-cultured with macrophages to analyze their effects on macrophages.Twenty nude mice(SPF grade,4 weeks old,15～19 g,half male and half female)were randomly divided into 4 groups(n=5):NC,INKA2-AS1,siNC and siINKA2-AS1.Tumor-bearing nude mouse models were established by subcutaneous injection of INKA2-AS1 silenced and overexpressed Hepa1-6 cells.The effect of INKA2-AS1 on tumor volume and mass were detected.Macrophage immunity was analyzed by Western blot analysis of IL-10 and FAP protein levels in tumor tissues.Results The level of INKA2-AS1 in HCC tissues was significantly higher than that in normal tissues(P<0.05).INKA2-AS1 was associated with the histological grade,pathological stage of HCC(P<0.05).The survival rate(P=0.05),disease-specific survival(P=0.036)and progression-free interval(P=0.024)of HCC patients with high INKA2-AS1 expression were significantly lower than those of patients with low INKA2-AS1 expression.The level of INKA2-AS1 in HCC tissues was significantly higher than that in adjacent normal tissues(P<0.05).Cox regression analysis showed that INKA2-AS1 was an independent predictor of OS in HCC patients(P=0.005).INKA2-AS1 expression was associated with immune cells such as T cells and macrophages.The proliferation,migration,and invasion abilities of the siINKA2-AS1 group were lower than those of the siNC group(P<0.05),the apoptotic rate was higher than that of the siNC group(P<0.05),and inhibited the expression of IL-10 and FAP at mRNA and protein levels in co-cultured macrophages(P<0.05).The proliferation,migration and invasion abilities of the INKA2-AS1 group were higher than those of the NC group(P<0.05),the apoptotic rate was lower than that of the NC group(P<0.05),and promoted the expression of IL-10 and FAP at mRNA and protein levels in co-cultured macrophages(P<0.05).The tumor volume and mass of the siINKA2-AS1 group were lower than those of the siNC group(P<0.05),while the tumor volume and mass of the INKA2-AS1 group were higher than those of the NC group(P<0.01).The levels of IL-10 and FAP proteins in tumor tissues of the siINKA2-AS1 group were lower than those of the siNC group(P<0.05),while the levels of IL-10 and FAP proteins were increased(P<0.05).Conclusion INKA2-AS1 increment is related to poor prognosis of HCC patients,and the molecule promotes the proliferation and metastasis of HCC cells and induces the transformation of tumor-associated macrophages.

Objective To investigate the mechanism by which SLC38A1 promotes hepatocellular carcinoma(HCC)progression through glutamine transport-mediated activation of the PI3K/AKT signaling pathway.Methods Bioinformatics analysis was employed to assess the correlation between SLC38A1 expression and clinicopathological features/prognosis in HCC patients,with functional enrichment analysis of SLC38A1 conducted.SLC38A1 was silenced or over expressed via transfection in Huh-7 cells,with glutamine inhibited using DRP-104 and the PI3K/AKT pathway suppressed using LY294002.Cell viability,proliferation,migration,invasion,and glutamine concentration were evaluated using CCK-8,EdU staining,scratch wound healing assay,Transwell chamber assay,and glutamine assay kits,respectively.PI3K/AKT pathway activity was assessed by Western blotting.Results SLC38A1 mRNA and protein levels were significantly higher in HCC tumor tissues than in adjacent normal tissues(P<0.05).HCC patients with high SLC38A1 expression exhibited significantly lower overall survival than those with low expression(P<0.05).SLC38A1 expression correlated significantly with pathologic T-stage(pT),N-stage(pN),M-stage(pM),survival status,and immune infiltration in HCC patients(P<0.05).SLC38A1 silencing markedly reduced glutamine uptake in HCC cells(P<0.001),suppressing cell viability,proliferation,migration,invasion,and PI3K/AKT pathway activity(P<0.001).Conversely,SLC38A1 overexpression promoted proliferation,migration,invasion,and PI3K/AKT activation(P<0.001).DRP-104-mediated glutamine inhibition suppressed HCC cell malignancy and PI3K/AKT signaling while abolishing the oncogenic effects of SLC38A1 overexpression(P<0.001).PI3K/AKT pathway inhibition blocked the pro-tumorigenic effects of SLC38A1 overexpression(P<0.001).Conclusion SLC38A1 promotes proliferation,migration,and invasion in HCC by activating the PI3K/AKT pathway through enhanced glutamine transport.

Objectives:To establish animal model of anterior proliferative vitreoretinopathy (aPVR) with cultured homologous dermal fibroblasts of rabbit, and to provide evidence why hypotony was caused by aPVR. Methods:Animal models of aPVR were established with cultured homologous dermal fibroblasts on pigmented rabbits. Rabbits were sacrificed on the 14th, 28th and 56th day after the operation to prepare naked eyes and to receive histological examinations. Results:Naked eye examination showed that the peripheral retina was detached by dragging in the experimental group 28 and 56 days postoperatively. Microscopic examination showed atrophy or absence of the non-pigmented ciliary epithelium on the 28th and 56th postoperative day in the experimental group. Conclusions:The epiciliary membrane in aPVR dragged the ciliary body, made atrophy of non-pigmented epithelium, which perhaps was the main cause of hypotony.

Objective Through transmission electron microscopy we observe the ultrastructure of optic nerve to evaluate the curative effect of optic nerve incision decompression (ONID) for incomplete injury to optic nerve. Methods The optic nerve was crushed for 30s to produce incomplete injury to the optic nerve in rats, and then the rats were divided into two groups. ONID was performed 6 hours after the injury in one group, and in the other group decompression was omitted(non-ONLD). Ultrastructure of optic nerve was observed 1 month, 2 months, 3 months and 6 months later in both groups. Results After 1 month denaturalization of myelin sheaths and axons of the injured optic nerve was severe in non- ONID group, and after 3 months the optic nerve entirely lost its normal structure. On the other hand, denaturalization of myelin sheaths and axons was evidently less intense in ONID group, and the changes in structure of optic nerve were stabilized. Conclusions ONID reduced the speed of degeneration of the injured optic nerve and postpones the process of axon denaturalization, thus protects the optic nerve from secondary injuries.

Objective To investigate the occurrence, progress and conversion of hypotony in anterior proliferative vitreoretinopathy (aPVR), and to provide knowledge about how to prevent and treat it. Methods Animal models of chronic hypotony by aPVR were made with cultured homologous dermal fibroblasts on pigmented rabbits. The intraocular pressure (IOP) and ultrasound biomicroscopy(UBM) examination were taken preoperatively and on days 7,14, 28 and 56 postoperatively. Rabbits were killed on days 14, 28 or 56 postoperatively, prepared for histology and ultrastructure examination. Results The average IOP of experimental group was lower than that of control group on days 7,14,28 and 56 significantly (P

Objective To examine the expression of proliferating cell nuclear antigen (PCNA) of retinal pigment epithelial (RPE) cells, thus assessing the role of mechanism of contact inhibition playing in the process of experimental retinal detachment and reattachemnt. Methods Retinal detachment was produced in 72 cats by subretinal injection of 0.25% solution of healon through a micropipette three weeks after extracapsular lens extraction and vitrectomy. Some of the detached retinae were reattached 24 hours later. At different time, the cats were killed and eye globes were fixed and embeded in paraffin. Histologic sections were processed for immunohistochemistry examination using an antibody to detect PCNA protein. Labeled RPE cells were identified, and the proliferation was quantified in detached and un detached retinae of detachment group, and also in reattached retinae of reattachment group. The comparsion of PCNA labeled RPE cells in different groups were analyzed by ANOVA. Results In detached regions of detachment group, PCNA expression of RPE cells occured within 24 hours, and reached a maximum after 5 6 days, then gradually declined to barely detectable levels after 20 days. Similar tendency was found in reattached retinae, but the number of PCNA labeled RPE cells was obviously small. Fewer PCNA labeled RPE cells were found in regions of un detached retinae in detachment group. The difference of these three groups was significant. Conclusion Proliferation of RPE cells is induced when they lose contact with neural retina, but inhibited after neural retina reattached to RPE cells. It suggests that the mechanism of contact inhibition plays a role in the proliferative process after retinal detachment and reattachment.

OBJECTIVE To investigate influence of compound gentamicin-fluorometholone(GentaFluoro) and tobramycin dexamethasone(TobraDex) eyedrops for treating ocular inflammation after cataract phacoemulsification.METHODS Sixty eyes in 60 patients after cataract phacoemulsification were equally randomized into two groups,such as GentaFluoro group and TobraDex group according to order of operation.Two groups were used respectively the two sort of eyedrops: GentaFluoro and TobraDex were used 6 times per day during first three days after the operation,later four days were four times per day.RESULTS GentaFluoro group and TobraDex group had identical results in controlling inflammation of anterior chamber. CONCLUSIONS The effects of GentaFluoro and TobraDex eyedrops for treating ocular inflammation after phacoemulsification are reliable.

To investigate the effectiveness and safety of systemic administration of high dose urokinase for acute central retinal artery occlusion (CRAO). 6 patients who had unilateral CRAO with symptoms lasting 6 hours to six days' (average, 3 7 days) received intravenous urokinase, 500,000U / day, with a total dose of 2,500,000U. Each urokinase delivery was followed by in travenous low molecular weight Dextran, 500ml / day, and Ginaton 87 5 mg / day, for 10 days. The treatment also included anterior chamber paracentesis, ocular massage, oral acetazolamide, sublingual isosorbide dinitrate, and inhalation of carbogen ( 95% oxygen, 5% carbon dioxide ). Visual acuity and ocular findings were recorded before and after treatment. Duration of follow up ranged from 4 to 12 months. After first intravaneous urokinase, visual acuity was slightly improved in five of the 6 patients, and no change in one patient. All 6 patients showed markedly improved vision at discharge, with vision better than 0 05. At the final visit, visual acuity reached 0 1 in all the 6 patients, and in four patients visual acuity was improved to 0 2 or better, two patients had vision recovered even to 1 2 and 1 5, respectively. During treatment no serious complications were noticed. These results indicate that systemic treatment for acute central retinal artery occlusion with high dose urokinase could help reestablish retinal circulation and improve vision.

Objective To explore the characteristic and the rule of flash-visual evoked potentials (F-VEP) of SD rats after optic nerve injury in different grades. Methods By using APS-2000 multifunctional electric physiological instrument, changes in F-VEP were observed in normal optic nerve and injured optic nerve (crushed in 3 grades) in rats. The latency and amplitude of F-VEP wave, were recorded and statistically. Results The F-VEP wave was reliably displayed with characteristic curve. The L5b was(53.67?3.12)ms and A5 was(17.83?5.91)?v. The latency of F-VEP in rats with optic nerve injury in different grades showed a significant prolongation, and the amplitude showed a significant decrease compared with normal rats (P

Objective To study the dynamics of aqueous humor in chronic hypotony induced by traumatic anterior proliferative vitreoretinopathy (aPVR), and to demonstrate physiologic mechanisms of the hypotony. Methods A model of hypotony to simulate traumatic aPVR was reproduced in rabbits. Preoperatively and on day 7, 14, 28 and 56 postoperatively, the aqueous humor flow rate and the uveoscleral outflow of aqueous humor were determined. Results The flow rate of aqueous humor in experimental group was reduced remarkably compared with that of control group on days 14, 28 and 56 (P