Short-term Evaluation of Visual Quality, Amplitude of Accommodation, and Stereoacuity Between Patients With Moderate-to-High Myopia Who Underwent ICLV4c Implantation and SMILE.

PURPOSE: To compare the visual quality, accommodation, and stereoacuity between patients with a myopia magnitude of -3.00 to -8.50 diopters (D) who underwent ICLV4c (STAAR Surgical) implantation and small incision lenticule extraction (SMILE). METHODS: The visual quality parameters, amplitude of accommodation (AMP), and stereoacuity were compared between and within the ICLV4c and SMILE groups (40 eyes/group) with a mean spherical equivalent (SE) of -6.14 D. Differences in these parameters before and after surgery were analyzed using SPSS 22.0 software (SPSS, Inc). RESULTS: Modulation transfer function (MTF) cut-off increased significantly by 3.24 cycles/degree (t = -2.111, P = .041) and AMP decreased significantly by 1.66 D (t = 7.519, P < .001) from baseline to 3 months after surgery in the ICLV4c group. However, the MTF cut-off was significantly lower (t = 2.123, P = .037) and the AMP was significantly better (t = -3.605, P = .001) in the SMILE group than in the ICLV4c group at 3 months postoperatively. Other parameters such as Objective Scatter Index, Strehl ratio, and Optical Quality Analysis System (Visiometrics) under different contrast values did not show significant changes (P > .05). Stereoacuity did not improve significantly, except in a patient with anisometropia (2.50 D) in the ICLV4c group. CONCLUSIONS: The ICLV4c implantation yielded a better visual quality but worse AMP than did SMILE in patients with a myopia magnitude of -3.00 to -8.50 D in the short term. [J Refract Surg. 2022;38(10):632-640.].

Astragaloside-IV alleviates high glucose-induced ferroptosis in retinal pigment epithelial cells by disrupting the expression of miR-138-5p/Sirt1/Nrf2.

Astragaloside-IV (AS-IV) (C41H68O14) is a high-purity natural product extracted from Astragalus, which has demonstrated biological activities. However, the effect of AS-IV on retinal pigment epithelial (RPE) cells in diabetic retinopathy (DR) remains unclear. In this study, high glucose (HG) was shown to promote ARPE-19 RPE cell death, increase the contents of reactive oxygen species (ROS) and oxidized glutathione (GSSG), and enhance lipid peroxidation density of mitochondrial membrane. In contrast, AS-IV decreased glutathione (GSH) content, mitochondria size and ridge. Addition of iron death inhibitor Ferrostatin-1 (Fer-1) to RPE cells decreased cell dead rate, thus indicating that HG-induced mitochondrial damage occurred due to ferroptosis. AS-IV alleviated HG-induced RPE cell damage. Furthermore, HG decreased levels of silent information regulator 1 (Sirt1) and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) in the nucleus of RPE cells; AS-IV could alleviate these effects and increased expression of glutathione peroxidase 4 (GPX4), glutamate cysteine ligase (GCLM) and glutamate cysteine ligase catalytic subunit (GCLC), which are Nrf2 downstream genes. Mechanistically, AS-IV was shown to alleviate the effects of HG by increasing mir-138-5p expression in RPE cells and promoting expression of Sirt1 and Nrf2 in the nucleus. Transfection of mir-138-5p agonist inhibited the regulatory effects of AS-IV on Sirt1 and Nrf2, accompanied by decreased GPX4, GCLM and GCLC levels, and restoration of ferroptosis-related changes. Collectively, HG increased ferroptosis rate in RPE cells. In addition, AS-IV inhibited miR-138-5p expression, subsequently increasing Sirt1/Nrf2 activity and cellular antioxidant capacity to alleviate ferroptosis, resulting decreased cell death, which potentially inhibits the DR pathological process.

Effects of dietary starch sources on pellet-processing characteristics, growth performance and caecal microflora of meat rabbits.

This study evaluated the effects of starch sources on pellet-processing characteristics as well as the growth performance and caecal microflora of rabbits. Ninety-six 35-day-old rabbits were randomly allocated to four groups with 24 rabbits per group and were fed diets with different starch sources (corn, wheat, potato or pea starch). The trial lasted for 40 days. The greatest hardness and lowest powder ratio of feed pellets was associated with the use of potato starch (p > 0.05). Pellet bulk density was the highest with corn starch, and the density was greater than that of pea starch by 5.91% (p < 0.05). The pulverisation ratio of corn starch pellets was the lowest, 43.67% lower than that of the pea starch pellets (p < 0.05). The average daily gain of rabbits in the corn starch group was higher than in the potato and pea starch groups, by 7.89% and 10.81%, respectively (p < 0.05). Rabbits in the corn starch group had the best feed conversion ratio (p > 0.05). The feed intake of rabbits in the potato starch group was higher than in the wheat and pea starch groups, by 4.30% and 5.16% respectively (p < 0.05). The dominant caecal bacteria phyla were Firmicutes, Bacteroidetes, Verrucomicrobia and Proteobacteria. There were 12 bacterial genera with proportions greater than 0.1%. The caecal proportion of Clostridium in the pea starch group was 1.8%, which was higher than those of the other groups (p = 0.057). There was no significant difference in caecal microbial diversity among groups (p > 0.05). The highest microbial clustering effect was found in the corn starch treatment. In conclusion, the best pellet quality was found using potato starch; for rabbit growth, the optimal source was corn starch.

Cyanidin-3-O-Glucoside Attenuates Lipopolysaccharide-Induced Inflammation in Human Corneal Epithelial Cells by Inducing Let-7b-5p-Mediated HMGA2/PI3K/Akt Pathway.

The bacterial keratitis causes viability loss and apoptosis in the corneal epithelial cells (CECs). The cyanidin-3-O-glucoside (C3G) benefits visual system and also possess anti-bacterial and anti-inflammatory potentials. In the current study, the effects of C3G on human CECs (HCECs) against bacterial lipopolysaccharide (LPS)-induced disorders were assessed, and the mechanism driving the protective effect was explored by focusing on let-7b-5p-mediated HMGA2/PI3K/Akt pathway. The HCECs were incubated LPS of P. aeruginosa to induce inflammation and apoptosis, and then treated with C3G. The changes in cell viability, apoptosis, and inflammation were detected. Moreover, the effects of LPS and C3G on let-7b-5p level and HMGA2/PI3K/Akt pathway activity were also assessed. Thereafter, the HCECs were further transfected with let-7b-5p inhibitor to confirm its role in the vision-protective effects of C3G. The interaction between let-7b-5p and HMGA2 was verified with dual luciferase assay. The LPS treatment suppressed viability and induced apoptosis and inflammation in HCECs, which was associated with the down-regulated let-7b-5p level and up-regulated HMGA2/PI3K/Akt pathway activity. The impairments of LPS on HCECs were attenuated by C3G: the compound increased cell viability and inhibited apoptosis and inflammation. The C3G also induced let-7b-5p level and inactivated HMGA2/PI3K/Akt pathway. However, after the inhibition of let-7b-5p, the protective effects of C3G on HCECs against LPS were blocked. The results of dual luciferase assay showed the direct binding let-7b-5p to the promoter of HMGA2 gene. It was inferred that the C3G could ameliorate the LPS-induced disorders in HCECs. The effect depended on the induced level of let-7b-5p, which then inhibited HMGA2/PI3K/Akt pathway.

Integrative transcriptomics and proteomic analysis of extraocular muscles from patients with thyroid-associated ophthalmopathy.

Our study aimed to reveal the underlying pathologic mechanisms of thyroid-associated ophthalmopathy (TAO) by integrative transcriptomics and proteomic analysis of extraocular muscles (EOM). The study involved 11 TAO patients (clinical activity score ≤ 2) and 11 control donors. Total RNA was extracted from EOM samples of 5 TAO patients and 5 control individuals for gene microarray analysis to reveal differentially expressed genes. Concurrently, EOM samples from 3 TAO patients and 3 control individuals were lysed for quantitative proteomic analysis. Differentially expressed genes and proteins were identified, followed by functional and pathway enrichment analysis and protein-protein interaction network construction. Concordance between proteins and transcripts was examined, and functional annotations were conducted. Expressions of versican (VCAN) and lipocalin 1 (LCN1) in EOM samples from another 3 TAO patients and 3 control individuals were measured by western blotting. In total, 952 genes and 137 proteins were identified as differentially expressed, as well as 96 differentially expressed proteins without significantly changed mRNA abundance. Proteins mainly related to the composition (such as MYH1, MYH2, and MYH13) and contraction force (MYH3, MYH8, ACTN3, and TNNT1) of the muscle fibers were significantly up-regulated in EOM samples of TAO, as well as those (such as VCAN, MPZ, and PTPRC) associated with cell adhesion. In addition, differentially expressed proteins related to the components and metabolism of extracellular matrix (ECM) (such as COL1A1, COL1A2, COL2A1, VCAN, OGN, and DCN) were identified. Similarly, expressions of genes involved in cell adhesion and ECM metabolism were significantly different between EOM samples of TAO patients and controls. Western blotting verified that VCAN involved in ECM proteoglycans and diseases associated with glycosaminoglycan metabolism was markedly higher in EOM samples of TAO, whereas LCN1 was obviously decreased. In conclusion, this study demonstrated the significantly altered cellular components of EOM, muscle contraction, cell adhesion and ECM metabolism, which might be involved in the pathologic mechanisms and/or consequences of TAO.

Roles of Lentinula edodes as the pork lean meat replacer in production of the sausage.

The effects of substituting pork lean meat with Lentinula edodes (LE) on the physicochemical properties, amino acid content, cooking loss, texture, total phenolic content, antioxidant activity, microstructure, microbiological analysis, and sensory characteristics of sausages were evaluated. Five formulations were used in the production of sausages: the control (the pork lean meat formulation) and the four different samples in which LE substituted 25%, 50%, 75%, and 100% of pork lean meat. The results showed that LE improved the moisture, total dietary fiber, methionine, glutamic, cysteine, and total phenolic content; cooking loss; and antioxidant activity of the sausage. By contrast, LE reduced the levels of protein, ash, pH, as well as the energy level and texture of the sausage. No difference was observed between the treatments for fat content, water activity and microorganisms of sausages. In addition, LE led to slight darkening of the sausages. From the sensory point of view, all modified sausages were considered acceptable, and the pork lean meat with 25% substitution by LE exhibited best sensory characteristics. In a word, LE is a promising ingredient to partially replace the lean meat in sausages.

Illumination from light-emitting diodes (LEDs) disrupts pathological cytokines expression and activates relevant signal pathways in primary human retinal pigment epithelial cells.

Age-related macular degeneration (AMD) is the leading cause of blindness in the aged people. The latest systemic review of epidemiological investigations revealed that excessive light exposure increases the risk of AMD. With the drastically increasing use of high-energy light-emitting diodes (LEDs) light in our domestic environment nowadays, it is supposed to pose a potential oxidative threat to ocular health. Retinal pigment epithelium (RPE) is the major ocular source of pathological cytokines, which regulate local inflammation and angiogenesis. We hypothesized that high-energy LED light might disrupt the pathological cytokine expression of retinal pigment epithelium (RPE), contributing to the pathogenesis of AMD. Primary human RPE cells were isolated from eyecups of normal eye donors and seeded into plate wells for growing to confluence. Two widely used multichromatic white light-emitting diodes (LEDs) with correlated color temperatures (CCTs) of 2954 and 7378 K were used in this experiment. The confluent primary RPE cells were under white LEDs light exposure until 24 h. VEGF-A, IL-6, IL-8 and MCP-1 proteins and mRNAs were measured using an ELISA kit and RT-PCR, respectively. Activation of mitogen-activated protein kinases (MAPKs), Akt, Janus kinase (JAK)2 and Nuclear factor (NF)-κB signal pathways after LEDs illumination were evaluated by western blotting analysis. The level of reactive oxygen species (ROS) using chloromethyl- 2',7'-dichlorodihydrofluorescein diacetate. Inhibitors of relevant signal pathways and anti-oxidants were added to the primary RPE cells before LEDs illumination to evaluate their biological functions. We found that 7378 K light, but not 2954 K upregulated the VEGF-A, IL-6, IL-8 and downregulated MCP-1 proteins and mRNAs levels in a time-dependent manner. In parallel, initial activation of MAPKs and NF-κB signal pathways were also observed after 7378 K light exposure. Mechanistically, antioxidants for eliminating reactive oxygen species (ROS) and targeted inhibitors of MAPKs and NF-κB significantly blocked 7378 K light-induced changes of specific cytokines, respectively. Our findings suggest that 7378 K light, not 2954 K induced upregulation of VEGF-A, IL-6, IL-8 and downregulation of MCP-1 via ROS accumulation, activating MAPKs and NF-κB signal pathways.

Interferon-λ1 suppresses invasion and enhances autophagy in human osteosarcoma cell.

OBJECTIVE: The purpose of the present study was to determine whether type III IFN can modulate the autophagic response in human osteosarcoma cell. METHODS: Human osteosarcoma cell were treated with Interferon-λ1. We investigated that Interferon-λ1 could inhibit the invasive ability of osteosarcoma cells by Matrigel invasion assay. Autophagy were assessed by acridine orange staining, MDC staining and Transmission electron microscopy. RESULTS: In this study, we found that Interferon-λ1 could inhibit the invasive ability of osteosarcoma cells. Acridine orange staining and MDC staining showed that Interferon-λ1 triggered the accumulation of acidic vesicular and autolysosomes in osteosarcoma cell. The acridine orange osteosarcoma cell ratios were 3.6 ± 0.5%, 4.5 ± 0.8%, and 12.4 ± 1.7% after treatment with 1, 10, and 100 ng/mL Interferon-λ1 for 48 h. Osteosarcoma cell cells treated with 100 ng/mL Interferon-λ1 for 48 h developed autophagy some-like characteristics, including single or double-membrane vacuoles containing intact and degraded cellular debris. CONCLUSIONS: Interferon-λ1 could inhibit the invasive ability of osteosarcoma cells. Autophagy can be induced in a dose-dependent manner by treatment with Interferon-λ1 in osteosarcoma cell.

Potential candidate cells for constructing tissue-engineered lacrimal duct epithelium: a histological and cytological study in rabbits.

OBJECTIVE: Injury and deficiency of the lacrimal duct epithelium (LDE) can lead to a variety of lacrimal diseases. The purpose of this study was to characterize potential candidate cells for constructing a tissue-engineered LDE. METHODS: Different areas of the conjunctiva and lacrimal duct tissue were removed from male adult New Zealand white rabbits for histological evaluation. Hematoxylin and eosin staining and immunohistochemical staining of cytokeratin AE1+AE3, cytokeratin 4, Ki-67, and MUC5AC were observed by light microscopy. The surface morphologies of different epithelial tissues and cellular structures were examined using field-emission scanning electron microscopy and transmission electron microscopy. Epithelial cells were isolated from tissues and identified by specific markers. In vitro, proliferative ability and Western blot analyses of the proliferating cell nuclear antigen (PCNA) of different epithelial cells cultured in identical environments were investigated and compared. RESULTS: Histologically, the epithelial specific markers, cytokeratin AE1+AE3 and cytokeratin 4, were expressed in the conjunctiva epithelium and the LDE. Notably, highly proliferative cells stained with Ki-67 were concentrated under the epithelium in a dome structure of the posterior palpebral conjunctiva. Differentiated goblet cells were also found to a lesser extent in this region. Primary palpebral and fornical conjunctival epithelial cells (PFCECs), bulbar conjunctival epithelial cells (BCECs), and lacrimal duct epithelial cells (LDECs) were successfully separated from tissues. In vitro, rabbit PFCECs and LDECs grew faster and expressed more PCNA than BCECs. CONCLUSIONS: PFCECs are anatomically similar to LDECs. They also have similar morphological characteristics, immune phenotypes, and proliferation features. PFCECs are therefore potential candidate cells to replace LDECs in tissue engineering to treat lacrimal duct diseases.

Long-term ultrasound biomicroscopy observation of position changes of a copolymer posterior chamber phakic intraocular lens.

PURPOSE: To evaluate longitudinal changes in Implantable Collamer Lens phakic intraocular lens (pIOL) position after implantation. SETTING: Department of Ophthalmology, Zhejiang University, Hangzhou, China. DESIGN: Retrospective case series. METHODS: Myopic eyes that had pIOL implantation with a follow-up of at least 24 months were evaluated. Ultrasound biomicroscopy examinations were performed at each visit. RESULTS: The study enrolled 62 eyes (31 patients; 22 women, 9 men) ranging in age from 21 to 46 years. The manifest spherical equivalent ranged from -8.25 to -18.75 diopters. A significant increase (mean 36 µm±50 [SD]) in the endothelium-anterior pIOL distance occurred between 1 month and 3 months (P=.000); afterward, the distance decreased slowly (P>.05). The largest decrease (mean 47±17 µm) in central vault occurred between 1 month and 3 months (P=.009). The largest decrease (mean 21±14 µm) in peripheral vault occurred between 1 month and 3 months (P=.000). CONCLUSIONS: A significant increase in the endothelium-anterior pIOL distance occurred from 1 month to 3 months postoperatively, after which a slight decrease occurred over time. Central vault and peripheral vault had a tendency to decrease over time. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Autophagy activated by Toxoplasma gondii infection in turn facilitates Toxoplasma gondii proliferation.

Autophagy was found to play an antimicrobial or antiparasitic role in the activation of host cells to defend against intracellular pathogens, at the same time, pathogens could compete with host cell and take advantage of autophagy to provide access for its proliferation, but there are few articles for studying the outcome of this competition between host cell and pathogens. Therefore, the aim of our study was to investigate the relationship between autophagy activated by Toxoplasma gondii (T. gondii) and proliferation of T. gondii affected by autophagy in vitro. Firstly, human embryonic fibroblasts (HEF) cells were infected with T. gondii for different times. The monodansylcadaverine (MDC) staining, acridine orange (AO) staining, punctuate GFP-LC3 distribution, and transmission electron microscopy (TEM) assays were conducted, and the results were consistent in showing that gondii infection could induce autophagy. Secondly, HEF cells were infected with T. gondii and treated with autophagy inhibitor bafilomycin A1 or inducer lithium chloride for different times. Giemsa staining was conducted, and the results exhibited that T. gondii infection-induced autophagy could in turn promote T. gondii proliferation. Simultaneously, the results of Giemsa staining also revealed that autophagy inhibitor could reduce the number of each cell infected with T. gondii and inhibit T. gondii proliferation. In contrast, autophagy inducer could increase the number of each cell infected with T. gondii and encourage T. gondii proliferation. Therefore, our study suggests that T. gondii infection could activate autophagy, and this autophagy could in turn facilitate T. gondii proliferation in HEF cells for limiting nutrients.

Effects of white light-emitting diode (LED) light exposure with different correlated color temperatures (CCTs) on human lens epithelial cells in culture.

Cataract is the major cause for legal blindness in the world. Oxidative stress on the lens epithelial cells (hLECs) is the most important factor in cataract formation. Cumulative light-exposure from widely used light-emitting diodes (LEDs) may pose a potential oxidative threat to the lens epithelium, due to the high-energy blue light component in the white-light emission from diodes. In the interest of perfecting biosafety standards for LED domestic lighting, this study analyzed the photobiological effect of white LED light with different correlated color temperatures (CCTs) on cultured hLECs. The hLECs were cultured and cumulatively exposed to multichromatic white LED light with CCTs of 2954, 5624, and 7378 K. Cell viability of hLECs was measured by Cell Counting Kit-8 (CCK-8) assay. DNA damage was determined by alkaline comet assay. Intracellular reactive oxygen species (ROS) generation, cell cycle, and apoptosis were quantified by flow cytometry. Compared with 2954 and 5624 K LED light, LED light having a CCT of 7378 K caused overproduction of intracellular ROS and severe DNA damage, which triggered G2 /M arrest and apoptosis. These results indicate that white LEDs with a high CCT could cause significant photobiological damage to hLECs.

[The relationship among congenital Toxoplasma gondii infection, pregnancy outcome and T lymphocyte subsets in umbilical cord blood].

OBJECTIVE: To reveal the relationship among congenital Toxoplasma gondii (T. gondii) infection, T lymphocyte cell subsets in umbilical cord blood and pregnancy outcome. METHODS: 784 umbilical cord blood samples were collected and information of pregnancy outcomes was collected in a hospital of Hefei city, Anhui province during March 2009 to May 2010. T. gondii IgM antibodies in the sera were detected by ELISA. For all neonates infected with T. gondii and 10 healthy neonates, T lymphocyte cell subsets were detected by flow cytometry. RESULTS: According to the detection results of T. gondii IgM antibodies, 784 neonates were divided into infection group (21 neonates) and control group (763 neonates). The body weight and 1 min Apgar score of infection group were (3116.4 ± 352.6) g and (8.21 ± 1.26) points, respectively, which were statistically lower than control group ((3220.1 ± 242.3) g and (8.77 ± 1.61) points, respectively) (P < 0.01). The proportion of adverse pregnancy outcome of infection group was 19.0% (4/21), which was statistically greater than control group (4.8%, 37/763) (P < 0.01). The percentage of CD(3)(+) T lymphocyte cells in umbilical cord blood in infection group with and without adverse pregnancy outcomes were (64.51 ± 5.27)% and (64.32 ± 4.56)%, respectively, which were statistically lower than control group ((69.32 ± 4.32)%) (P < 0.01). The ratio value of CD(4)(+)/CD(8)(+) in infection group with, without adverse pregnancy outcomes and control group are 1.39 ± 0.24, 1.64 ± 0.28 and 2.34 ± 0.46, respectively, which showed statistical difference between any 2 groups (P < 0.01). CONCLUSION: T. gondii infection leads to adverse pregnancy outcomes and disorder of cellular immunity while T lymphocyte cell subsets are closely associated with adverse pregnancy outcome.

Indications and outcomes of penetrating keratoplasty in a tertiary hospital in the developing world.

BACKGROUND: To assess the indications for and graft outcomes of penetrating keratoplasty (PK) in a tertiary hospital in China. DESIGN: A retrospective analysis of hospital records of patients, who had undergone PK at the First Affiliated Hospital of Zhejiang University in China over a 10-year period, was performed. PARTICIPANTS: A total of 203 eyes met the inclusion criteria for this study. METHODS: All PKs were performed with a standard technique throughout. MAIN OUTCOME MEASURES: Outcome measures included best corrected visual acuity at last follow-up, and overall graft survival. Data were assessed by Kaplan-Meier survival methods, univariate analysis and multivariate cox proportional hazards regression modelling. RESULTS: Among 203 eyes included in this study, the most common indications for PK in this study group were corneal scar caused by herpes simplex keratitis (24.1%) and trauma (mechanical and chemical) (21.2%). At the last follow-up, significantly more patients (35.0%) achieved 20/40 or better after PK compared with preoperation (P < 0.05). Overall, the 2-year graft survival rate was 79.9 ± 3.6%. In a univariate analysis, gender, indications for PK, type of operative procedure, rejection episode, postoperative glaucoma and postoperative herpetic recurrence were shown to be significantly associated with graft survival. In a multivariate analysis, three independent predictors of graft failure were identified including indications, gender and rejection episode. CONCLUSIONS: PK is effectively performed to improve visual function in southeast China. The outcomes of this study concur with other published studies and can be used as prognostic guidelines for any tertiary hospital in developing world.

Genotype-phenotype correlations in Chinese patients with TGFBI gene-linked corneal dystrophy.

In this paper, we report the clinical and molecular features of the distinct TGFBI (human transforming growth factor ß-induced, OMIM No. 601692) gene-linked corneal dystrophy. Altogether, five pedigrees and ten unrelated individuals diagnosed as corneal dystrophy were recruited. Peripheral venous DNA was extracted, and then amplified by polymerase chain reaction (PCR) and scanned for mutation by single-stranded conformation polymorphism (SSCP). Direct DNA sequencing was used to analyze the mutations of the TGFBI gene. In our study, thirty patients from five pedigrees and ten sporadic patients were diagnosed as four TGFBI gene-linked corneal dystrophies of granular corneal dystrophy type I (GGCD I), Avellino corneal dystrophy (ACD), lattice corneal dystrophy type I (LCD I), and lattice corneal dystrophy type IIIA (LCD IIIA), and in total, seven disease-causing mutations, namely R555W, A546D, A546T, and T538P mutations in exon 12, R124H and R124C mutations in exon 4, and P501T mutation in exon 11, were identified, while four polymorphisms of V327V, L472L, F540F, and 1665-1666insC were screened in exons 8, 11, and 12. The study ascertained the tight genotype-phenotype relationship and confirmed the clinical and genetic features of four TGFBI gene-linked corneal dystrophies.

Anti-tumor effect of 125I-UdR in combination with Egr-1 promoter-based IFNγ gene therapy in vivo.

Although (125)I-UdR treatment of malignant tumors in animal models and patients has achieved a certain effect, the short half-life of (125)I-UdR in vivo and its cellular uptake only in S phase of the cell cycle are limiting factors with regard to tumor eradication, and therefore its combination with other applications is a promising strategy in cancer therapy. In this study, we show that (125)I-UdR radionuclide therapy in combination with Egr-1 promoter-based IFNγ gene therapy is more effective than (125)I-UdR therapy alone in suppressing tumor growth and extending survival duration in mice bearing H22 hepatomas. Combined therapy could significantly inhibit cell proliferation and tumor angiogenesis, induce apoptosis and enhance cytotoxic activities of splenic CTL of the mice. Our results suggest that (125)I-UdR in combination with Egr-1 promoter-based IFNγ gene therapy may provide novel approaches for cancer treatment.

Low-dose radiation induces adaptive response in normal cells, but not in tumor cells: in vitro and in vivo studies.

Biological effects of low-dose radiation (LDR) are distinguishable from those of high-dose radiation. Hormetic and adaptive responses are such two examples. However, whether adaptive response could be induced in tumor cells by LDR, especially under in vivo condition, remains elusive, and was systemically investigated in the present study. Four tumor cell lines: two human leukemia cell lines (erythroleukemia cell line K562, and acute promyelocytic leukemia cell line HL60), and two human solid tumor cell lines (lung carcinoma cell line NCI-H446 and glioma cell line U251), along with one normal cell line (human fibroblast cells, MRC-5), were irradiated with LDR at 75 mGy of X-rays as D1 and then 4 Gy of X-rays as D2 (i.e.: D1 + D2) or only 4 Gy of X-rays (D2 alone). Three tumor-bearing animal models were also used to further define whether LDR induces adaptive response in tumor cells in vivo. Adaptive response was observed only in normal cell line, but not in four tumor cell lines, in response to LDR, showing a resistance to subsequent D2-induced cell growth inhibition. Three tumor-bearing mouse models with U251, NCI-H446 or S180 tumor cells were used to confirm that pre-exposure of tumor-bearing mice to D1 did not induce the resistance of tumor cells in vivo to D2-induced tumor growth inhibition. Furthermore, a higher apoptotic effect, along with higher expression of apoptosis-related genes P53 and Bax and lower expression of anti-apoptosis gene Bcl-2, was found in tumor cells of the tumor-bearing mice exposed to D1 + D2 than those in the tumor cells of the tumor-bearing mice exposed to D2 alone. These results suggest that LDR does not induce adaptive response in the tumor cells under both in vitro and in vivo conditions, which is a very important, clinic-relevant phenomenon.

In vitro anti-HIV and -HSV activity and safety of sodium rutin sulfate as a microbicide candidate.

Sodium rutin sulfate (SRS) is a sulfated rutin modified from the natural flavonol glycoside rutin. Here, we investigated its in vitro anti-HIV and -HSV activities and its cytotoxic profile. Fifty percent inhibitory concentration (IC(50)) values of SRS against HIV-1 X4 virus IIIB, HIV-1 R5 isolates Ada-M and Ba-L were 2.3+/-0.2, 4.5+/-2.0 and 8.5+/-3.8 microM with a selectivity index (SI) of 563, 575 and 329, respectively. Its IC(50) against primary R5 HIV-1 isolate from Yunnan province in China was 13.1+/-5.5 microM, with a SI of 197. In contrast, unsulfated rutin had no activity against any of the HIV-1 isolates tested. Further study indicated that SRS blocked viral entry and virus-cell fusion likely through interacting with the HIV-1 envelope glycoprotein. SRS also demonstrated some activity against human herpes simplex virus (HSV) with an IC(50) of 88.3+/-0.1 microM and a SI of 30. The 50% cytotoxicity concentration (CC(50)) of SRS was >3.0 mM, as determined in human genital ME180, HeLa and primary human foreskin fibroblast cells. Minimum inhibitory concentration of SRS for vaginal lactobacilli was >3.0 mM. These results collectively indicate that SRS represents a novel candidate for anti-HIV-1/HSV microbicide development.

Effect of pEgr-TNFalpha gene radiotherapy on mice melanoma.

In the present study, we constructed a pEgr-tumour necrosis factor-alpha (TNFalpha) plasmid and investigated its expression properties in B16 cells on exposure to ionizing irradiation and, furthermore, the effect of gene radiotherapy on a melanoma model. Firstly, the recombinant pEgr-TNFalpha plasmid was constructed and transfected into B16 cells with liposomes to investigate its expression properties on exposure to X-irradiation. The melanoma-bearing model was then established and the tumour tissue was injected locally with the pEgr-TNFalpha plasmid and exposed to 20 Gy of X-irradiation. The tumour growth curve at different time points was described. At day 3, TNFalpha transcription in the tumour tissue was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that: (1) the eukaryotic expression vector pEgr-TNFalpha was successfully constructed and transfected into B16 cells; (2) TNFalpha expression was significantly increased in the transfected cells after X-irradiation, in contrast with that of the pCMV-TNFalpha group or the pEgr-TNFalpha group that received 0 Gy of irradiation; (3) after pEgr-TNFalpha gene radiotherapy, tumour growth was significantly slower than in those groups receiving irradiation or gene transfer alone. The TNFalpha concentration in the peripheral blood of tumour-bearing mice that received an injection of pEgr-TNFalpha and 20 Gy of irradiation was higher than that of the control group. Only in pEgr-TNFalpha and pEgr-TNFalpha + 20 Gy groups was TNFalpha messenger RNA (mRNA) detected in the tumour tissue. We conclude that pEgr-TNFalpha gene radiotherapy may significantly inhibit tumour growth, and that the anti-melanoma effect is superior to that of either gene therapy or radiotherapy alone. Our work provides the theoretical basis for further study on the gene radiotherapy of this tumour.