Nono deficiency impedes the proliferation and adhesion of H9c2 cardiomyocytes through Pi3k/Akt signaling pathway.

Congenital heart disease (CHD) is the most common type of birth defect and the main noninfectious cause of death during the neonatal stage. The non-POU domain containing, octamer-binding gene, NONO, performs a variety of roles involved in DNA repair, RNA synthesis, transcriptional and post-transcriptional regulation. Currently, hemizygous loss-of-function mutation of NONO have been described as the genetic origin of CHD. However, essential effects of NONO during cardiac development have not been fully elucidated. In this study, we aim to understand role of Nono in cardiomyocytes during development by utilizing the CRISPR/Cas9 gene editing system to deplete Nono in the rat cardiomyocytes H9c2. Functional comparison of H9c2 control and knockout cells showed that Nono deficiency suppressed cell proliferation and adhesion. Furthermore, Nono depletion significantly affected the mitochondrial oxidative phosphorylation (OXPHOS) and glycolysis, resulting in H9c2 overall metabolic deficits. Mechanistically we demonstrated that the Nono knockout impeded the cardiomyocyte function by attenuating phosphatidyl inositol 3 kinase-serine/threonine kinase (Pi3k/Akt) signaling via the assay for transposase-accessible chromatin using sequencing in combination with RNA sequencing. From these results we propose a novel molecular mechanism of Nono to influence cardiomyocytes differentiation and proliferation during the development of embryonic heart. We conclude that NONO may represent an emerging possible biomarkers and targets for the diagnosis and treatment of human cardiac development defects.

Comprehensive analysis of an autophagy-related prognostic model for predicting survival based on TCGA and ICGC database in hepatocellular carcinoma patients.

Background: There is accumulating evidence that autophagic activity is crucial to the development of hepatocellular carcinoma (HCC). Thus, we sought to develop a predictive model based on autophagy-related genes (ARGs) to forecast the prognosis of HCC patients. Methods: Based on expression data from The Cancer Genome Atlas (TCGA) and ARGs from Human Autophagy Database (HADb), the differentially expressed ARGs were screened. The prognosis-related ARGs were identified using a univariate Cox regression analysis. Using multivariate Cox regression analysis, a prognostic model was developed. To assess the predictive value of the model, receiver operating characteristic (ROC) curve, Kaplan-Meier curve, and multivariable Cox regression analyses were conducted. A data cohort gathered independently from the International Cancer Genome Consortium (ICGC) database further verified the model's predictive accuracy. The immune landscape was generated using the TIMER and CIBERSORT algorithms. Finally, the correlation between the prognostic signature and gene mutation status was analyzed by employing "maftools" package. Results: We identified a novel prediction model based on the ARGs of PLD1 and SLC36A1 with significant prognostic values for HCC in both univariate and multivariate Cox regression analysis, and patients were classified into high- or low-risk groups based on their risk scores. High-risk patients had significantly shorter overall survival (OS) times than low-risk patients (P=5e-4). According to the ROC curve analysis, the risk score had a higher predictive value than the other clinical characteristics. Prognostic nomograms were also performed to visualize the relationship between individual predictors and survival rates in patients with HCC. Further, an external independent cohort of ICGC patients provided additional confirmation of the predictive efficacy of the model. We subsequently analyzed the differential immune densities of the two groups and discovered that various immune cells, including naïve B cells, resting memory cluster of differentiation (CD)4 T cells, regulatory T cells, M2 macrophages, and neutrophils, had considerably larger infiltrating densities in the high-risk group than the low-risk group. Conclusions: We established a robust autophagy-related risk model having a certain prediction accuracy for predicting the prognosis of HCC patients. Our findings will contribute to the definition of prognosis and establishment of personalized treatment interventions for HCC patients.

Centromere-Specific Retrotransposons and Very-Long-Chain Fatty Acid Biosynthesis in the Genome of Yellowhorn (Xanthoceras sorbifolium, Sapindaceae), an Oil-Producing Tree With Significant Drought Resistance.

In-depth genome characterization is still lacking for most of biofuel crops, especially for centromeres, which play a fundamental role during nuclear division and in the maintenance of genome stability. This study applied long-read sequencing technologies to assemble a highly contiguous genome for yellowhorn (Xanthoceras sorbifolium), an oil-producing tree, and conducted extensive comparative analyses to understand centromere structure and evolution, and fatty acid biosynthesis. We produced a reference-level genome of yellowhorn, ∼470 Mb in length with ∼95% of contigs anchored onto 15 chromosomes. Genome annotation identified 22,049 protein-coding genes and 65.7% of the genome sequence as repetitive elements. Long terminal repeat retrotransposons (LTR-RTs) account for ∼30% of the yellowhorn genome, which is maintained by a moderate birth rate and a low removal rate. We identified the centromeric regions on each chromosome and found enrichment of centromere-specific retrotransposons of LINE1 and Gypsy in these regions, which have evolved recently (∼0.7 MYA). We compared the genomes of three cultivars and found frequent inversions. We analyzed the transcriptomes from different tissues and identified the candidate genes involved in very-long-chain fatty acid biosynthesis and their expression profiles. Collinear block analysis showed that yellowhorn shared the gamma (γ) hexaploidy event with Vitis vinifera but did not undergo any further whole-genome duplication. This study provides excellent genomic resources for understanding centromere structure and evolution and for functional studies in this important oil-producing plant.

Male reproductive systems of Macaca mulatta: Gonadal development, spermatogenesis and applications in spermatogonia stem cell transplantation.

Rhesus macaque (Macaca mulatta) is widely applied in animal model construction of infertility, spermatogonia stem cell transplantation and male reproductive diseases. In this review, we describe the seasonal changes of the reproductive system in rhesus macaques, the regular pattern of spermatogenesis and spermatozoa maturation, and the differentiation of spermatogonia and spermatocytes. The duration of the M. mulatta spermatogenesis is approximately 10 days and seminiferous epithelium cycles mainly consist of 12 stages, which provide a suitable model for reproductive studies in non-human primates. Here, we summarize the features of gonadal development and sperm maturation in the rhesus monkeys, which provide important information in the studies of reproductive biology. Rhesus macaque is an excellent animal model in spermatogonia stem cell transplantation. We discuss the applications and progresses of assisted reproductive technologies in sperm liquefaction, semen cryopreservation and spermatogonia stem cell transplantation of rhesus macaques. Besides, we sort out recent proteomic analyses of male reproductive systems and semen samples in rhesus macaques. This review mainly focuses on male reproductive biology and application studies using M. mulatta, which would promote the development of new therapeutic interventions on assisted reproduction and reproductive disease studies in the future.

Solvothermal preparation of luminescent zinc(II) and cadmium(II) coordination complexes based on the new bi-functional building block and photo-luminescent sensing for Cu2+, Al3+ and L-lysine.

In industry, over usage of Cu2+ and Al3+ will lead to toxic wastewater, which further to give serious pollution for the environment. On the other hand, L-lysine can enhance serotonin release in the amygdala, with subsequent changes in psychobehavioral responses to stress. Therefore it is the urgent problem to design a method for detecting the amount of Cu2+, Al3+, and L-lysine. In this work, through the solvothermal synthesis method, two new coordination complexes based on the new bifunctional building block 4'-(1H-1,2,4-triazole-1-yl)- [1,1'-biphenyl]-4-carboxylic acid (HL) have been synthesized, namely, [Zn(L)2·4H2O] (complex 1) and [Cd(L)2·4H2O] (complex 2). X-ray single-crystal diffractometer was used to analyze its structure, powder X-ray diffraction (PXRD) patterns confirmed that 1 and 2 powder's purity and 1 can keep stable during the detection process of Cu2+, Al3+, and L-lysine, respectively. Elemental analysis, thermogravimetric analysis, infrared analysis, ultraviolet analysis and fluorescent spectrum have been used to characterize these complexes. The photo-luminescent test showed that 1 can accurately recognize Al3+ and Cu2+ among various cations. On the other hand, 1 can distinguish L-lysine among amino acid molecules. Therefore, 1 can be utilized as a multifunctional fluorescent probe for Al3+(Ksv = 1.5570 × 104 [M]-1), Cu2+(Ksv = 1.4948 × 104 [M]-1) and L-lysine (Ksv = 4.9118 × 104 [M]-1) with low detection limits (17.5 µM, 18.2 µM, 5.6 µM) respectively.

A recognition survey of granular corneal dystrophy type 2 genetic detection in China.

AIM: To evaluate the feasibility of promoting genetic detection for granular corneal dystrophy type 2 (GCD2) by a questionnaire conducted among citizens in five cities in China. METHODS: The data were collected by questionnaire, and analyzed by Chi-square test and one-tailed t test in IBM SPSS statistics. RESULTS: Based on the survey data on the awareness of GCD2 genetic detection in this study and the positive predictive analysis report of the citizens in five cities in China, the vast majority (84.2%) of respondents had never heard of it and did not know that GCD2 patients have been prohibited from performing excimer surgery that can deteriorate GCD2 patients' condition even leading to blindness. Though 3.4% of patients understood GCD2 very much, they have no idea that GCD2 could not be 100% accuracy diagnosed by the conventional inspection methods. CONCLUSION: It is feasible and necessary to use GCD2 genetic detection as an excimer preoperative examination project. In order to promote the development of detection project, a few improvements should be carried out in terms of the promoting efforts, costs, and research progress.

Berberine-loaded Janus gold mesoporous silica nanocarriers for chemo/radio/photothermal therapy of liver cancer and radiation-induced injury inhibition.

Background: The combination of chemotherapy with radiotherapy serves as a common therapeutic strategy in clinics. However, it is unsatisfactory due to its poor therapeutic efficiency and severe side-effects originating from chemotherapy-exerted systemic toxicity as well as radiation-induced injury. Purpose: Hence, Berberine (Ber), an isoquinolin alkaloid with low toxicity and protective effects against radiotherapy, was used as a novel chemotherapeutic agent for chemo-radiotherapy of liver cancer. Patients and methods: We preloaded Ber into folic acid targeting Janus gold mesoporous silica nanocarriers (FA-JGMSNs) for overcoming the poor bioavailability of Ber. Furthermore, FA-JGMSNs were not only employed as radiosensitizers for expanding radiotherapeutic effect, but also used as photothermal agents for supplementing chemo-radiotherapeutic effect by local photothermal therapy. Results: In vitro and in vivo experiemtal results demonstrated the highly efficient anti-tumor effect, good biosafety as well as the effective protection of normal tissue of this nanoplatform. Conclusion: Based on its superb performance, we believe our work provided a feasible strategy for triple-therapies of liver cancer.

Anion directing self-assembly of 2D and 3D water-stable silver(i) cation metal organic frameworks and their applications in real-time discriminating cysteine and DNA detection.

Two water-stable silver(i) cation metal organic frameworks (MOFs), namely 2D MOF {[Ag(L)2]BF4}n (1) and 3D MOF {[Ag3(L)3]·(H2O)·(CF3SO3)3}n (2) (L = 1-(4-aminobenzyl)-1,2,4-triazole), have been prepared. 1 is the first example of a 2D MOF-based sensor for real-time discrimination of l- and d-cysteine from other amino acids through the quenching effect. Through deliberately tuning the nanoparticles of 2 under ultrasound conditions, these nanoparticles are, for the first time, successfully applied as an excellent DNA detecting platform.

Different phenotypes of monocytes in patients with new-onset mild acute pancreatitis.

AIM: To evaluate the numbers of different subsets of monocytes and their associations with the values of clinical measures in mild acute pancreatitis (MAP) patients. METHODS: The study included one group of 13 healthy controls and another group of 24 patients with new-onset MAP. The numbers of different subsets of monocytes were examined in these two groups of subjects by flow cytometry. The concentrations of plasma interleukin (IL)-10 and IL-12 were determined by cytometric bead array. The acute physiology and chronic health evaluation (APACHE) II scores of individual patients were evaluated, and the levels of plasma C-reactive protein (CRP) as well as the activities of amylase and lipase were measured. RESULTS: In comparison with that in the controls, significantly increased numbers of CD14+CD163-, CD14+CD163-MAC387+ M1 monocytes, but significantly reduced numbers of CD14+CD163+IL-10+ M2 monocytes were detected in the MAP patients (P < 0.01 or P < 0.05). Furthermore, significantly higher levels of plasma IL-10 and IL-12 were observed in the MAP patients (P < 0.01 for all). More importantly, the levels of plasma CRP were positively correlated with the numbers of CD14+CD163- (R = 0.5009, P = 0.0127) and CD14+CD163-MAC387+ (R = 0.5079, P = 0.0113) M1 monocytes and CD14+CD163+CD115+ M2 monocytes (R = 0.4565, P = 0.0249) in the patients. The APACHE II scores correlated with the numbers of CD14+CD163+CD115+ (R = 0.4581, P = 0.0244) monocytes and the levels of plasma IL-10 (R = 0.4178, P = 0.0422) in the MAP patients. However, there was no significant association among other measures tested in this population. CONCLUSION: Increased numbers of CD14+CD163- and CD14+ CD163-MAC387+ monocytes may contribute to the pathogenesis of MAP, and increased numbers of CD14+CD163+CD115+ monocytes may be a biomarker for evaluating the severity of MAP.

[Meta-analysis of internal fixation versus hip replacement in the treatment of trochanteric fractures].

OBJECTIVE: To compare the efficacy of internal fixation (including PFNA and PFN) versus hip replacement (including FHR or THA) in the treatment of trochanteric fractures in adults. METHODS: Reports of studies using randomized controlled trials (RCT) to compare internal fixationg with hip replacement in the management of intertrochanteric fractures were retrieved (up to January 1, 2013) from the Cochrane Library, PUBMED Data, CNKI (China National Knowledge infrastructure), Elsevier, the Chinese Biomedical Database, Wanfang Data, and manually. Methodological quality of the trials was critically assessed, and relevant data were extracted. Statistical software RevMan 5.0 was used for data-analysis. RESULTS: Seven articles were included in the meta-analysis. The results showed that,compared internal fixation with hip replacement,there were statistical significance in the duration of surgery time [WMD = -2.66, 95% CI (-5.25,-0.06), P = 0.05], intra-operative blood loss [WMD = -24.20, 95% CI (-30.38, -18.02), P < 0.000 01], hospital stays time [WMD = -4.72, 95% CI (-5.18, -4.25), P < 0.000 01], bearing load time [WMD = -29.54, 95% CI (-30.77, -28.31), P < 0.000 01], total complications rate [WMD = 0.15, 95% CI (0.11, 0.22), P < 0.000 01], the good rate of Harris scores [WMD = 1.09, 95% CI (0.54,1.32), P < 0.05]. However, there were no statistical significance in the rate of deep venous thrombosis [WMD = 1.09, 95% CI (0.47, 2.55), P > 0.05]. CON- CLUSION: Hip replacement (containing FHR or THA) for the treatment of intertrochanteric fractures is superior to internal fixa- tion in regards to the duration of surgery time, the mean duration of hosipital stays, mean post-operative down time, intra-opera- tive blood loss, the rate of post-operative good Harris scores. But there is not enough evidence to show any difference between hip replacement (containing THA or FHR) and internal fixation in regards to the rate of deep venous thrombosis. However, internal fixation for the treatment of intertrochanteric fractures is superior to hip replacement (containing FHR or THA) in regards to total complications rate.

Effect of bone marrow stromal cell transplantation on neurologic function and expression of VEGF in rats with focal cerebral ischemia.

There is evidence that the transplantation of mesenchymal stem cells into rat models of cerebral ischemia reduces ischemic damage; however, the mechanism remains to be elucidated. The present study aimed to assess the effect of transplantation of human bone marrow stromal cells (hBMSCs) on neurologic function and the expression of vascular endothelial growth factor (VEGF) in a rat model of focal cerebral ischemia. The left middle cerebral artery of adult Wistar rats was occluded for 90 min using a nylon thread, followed by reperfusion for 1 h. hBMSCs labeled with 5-bromo-2-deoxyuridine (BrdU) were stereotaxically injected into the ischemic boundary zone. Behavioral analysis using the Neurological Severity Score (NSS) was conducted on days 1, 3, 7 and 28, and a histologic evaluation was performed simultaneously. VEGF was detected by immunofluorescence staining and western blot analysis. Fifty rats were divided equally into five groups: Normal control, sham­operated, operated (no transplantation), Dulbecco's medium Eagle's medium (DMEM)-injected (received only serum-free DMEM), and hBMSC-transplanted. The hBMSC-transplanted group showed significantly improved behavioral recovery compared with the operated and DMEM-transplanted groups on days 3, 7 and 28. Histological examination showed that transplanted cells migrated from the injection site into nearby areas including the cortex. Expression of VEGF was significantly greater in the hBMSC group compared with the other four groups on each assessment day. The expression of VEGF was found to be beneficial for functional recovery following cerebral ischemic injury and hBMSC transplantation stimulated the expression of VEGF. Transplantation of BMSCs may be a promising therapeutic strategy for treating cerebral infarction.

[Effect of bone marrow mesenchymal stem cells on tumor neovascularization].

OBJECTIVE: The effect of human bone marrow mesenchymal stem cells (hMSCs) on tumor neovascularization were studied. METHODS: hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. hMSCs-EGFP were obtained by pLEGFP-N1 retroviral vector. Flow cytometry was used to detect the cell surface antigen and the differentiation potential of hMSCs-EGFP was investigated under conditioned media. The effect of hMSCs on tumor neovascularization were observed by establishing solid tumor models in BALB/C nude mice. In addition, effect of the conditioned medium used for tumor cells and endothelial cells (EC) cultivation was collected, to detect its effect on the growth and migration rates of hMSC. hMSCs were induced to differentiate into EC in vitro and the migratory effect on HUVEC was also evaluated. RESULTS: hMSCs-EGFP, like hMSC, exhibited a fibroblast-like morphological feature, and both had the similar cell surface antigens. They could be induced into osteocytes or adipocytes under the conditioned media. The results not only suggested that hMSCs contributed to tumor neovascularization, but also indicated that most of vessels were host-derived angiogenesis mediated by hMSCs. The mean vascular density (MVD) in suspension group (13.67 ± 1.53) was strikely higher than that in MCF-7 group (5.33 ± 1.42), which showed statistical significance (P < 0.05). Only very few vessels were attributed to hMSCs transdifferentiation into ECs. Tumor cells and ECs can promote hMSCs proliferation and migration through paracrine action. Furthermore, hMSCs were positive for CD31 after 2 weeks induction and HUVEC migration can be facilitated by hMSCs. CONCLUSION: MSCs have the effect of promoting tumor neovascularization.

[Role of aquaporin 1 in the migration of eosinophils from asthmatic guinea pigs].

OBJECTIVE: To investigate the role of aquaporin 1 (AQP1) in the migration of eosinophils (EOS) and to determine if AQP-1 can be viewed as the chemotactic activity marker of EOS. METHODS: Asthma model of guinea pigs were developed and EOS were purified from both peripheral blood and bronchoalveolar lavage fluid (BALF). The smears of EOS were studied by in situ hybridization for determining AQP1 mRNA and immunofluorescence under laser scanning confocal microscope for determining AQP1 protein. RESULTS: AQP1 was found expressed in EOS both from peripheral blood and from BALF. Compared with the expression of AQP1 mRNA (mean grey value 109.200 +/- 5.756, x +/- s) and protein (average fluorescence intensity 279.926 +/- 11.293) in EOS from BALF, there was stronger expression of AQP1 mRNA (92.904 +/- 3.290) and protein (425.081 +/- 17.474) in EOS from peripheral blood. The difference both of AQP1 mRNA (t = 9.519, P < 0.05) and protein(t = 27.020, P < 0.05) were considered statistically significant respectively. CONCLUSIONS: AQP1 plays a crucial role in EOS movement. It is possible that EOS produce more AQP1 protein to accelerate its migration to inflammatory tissues under allergic disease and EOS with AQP1 highly expressed are activated. AQP1 can be viewed as the chemotactic activity marker of EOS.

[Mesenchymal stem cells transduced by PLEGFP-N1 retroviral vector maintain their biological features and differentiation].

BACKGROUND: Enhanced green fluorescent protein (EGFP) has been an important reporter gene for gene therapy. Human mesenchymal stem cells (hMSCs) are ideal target cells in cell transplantation and tissue engineering. We investigated their biological characteristics and differentiation mediated by PLEGFP-N1 retroviral transduction. METHODS: hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. Individual colonies were selected and cultured in tissue dishes. Packaging cells PT67 were transfected by PLEGFP-N1 retroviral vector, and hMSCs were transduced by viral supernatant infection. Meanwhile, hMSCs-EGFP were identified by immune phenotypes and whether it could differentiate into osteoblasts or adipocytes under conditioned media was investigated. RESULTS: The rate of stably transduced hMSCs-EGFP was up to 96% after being screened by G418. hMSCs-EGFP exhibited fibroblast-like morphological features. Flow cytometric analyses showed that hMSCs-EGFP were positive for CD73, CD105, CD166, CD90 and CD44, but negative for CD34 and CD45. In addition, it could functionally be induced into osteocytes or adipocytes under conditioned media. These biological features of hMSCs-EGFP were consistent with those of hMSCs. CONCLUSIONS: hMSCs transduced by PLEGFP-N1 retroviral vector can be used in vivo securely because they can maintain their biological characteristics and differentiation. It is a simple and reliable way to trace the changes of hMSCs in vivo by EGFP during cell transplantation and gene therapy.

Inhibition of vascular endothelial growth factor gene expression by T7-siRNAs in cultured human retinal pigment epithelial cells.

BACKGROUND: Retinal pigment epithelial (RPE) cells play an important role in the occurrence of choroidal neovascularization (CNV). Vascular endothelial growth factor (VEGF) as a positive regulatory growth factor is produced by the RPE in an autocrine or paracrine manner, promoting CNV development. Duplexes of 21 nt RNAs, known as short interfering RNAs (siRNAs), efficiently inhibit gene expression by RNA interference when introduced into mammalian cells. We searched for an efficient siRNA to interfere with VEGF expression in RPE cells and shed light on the treatment of CNV. METHODS: Human primary RPE (hRPE) cells were cultured and identified. Three pairs of siRNAs were designed according to the sequence of VEGF 1-5 extrons and synthesized by T7 RNA polymerase transcription in vitro. To evaluate the inhibitory activity of T7-siRNAs, hRPE cells were transfected via siPORT Amine. The interfering effect of T7-siRNAs in hRPE cells was examined by semiquantitative reverse transcription-polymerase chain reaction and immunofluorescence. RESULTS: Three pairs of T7-siRNAs synthesized by in vitro transcription with T7 RNA polymerase suppressed VEGF gene expression with efficiency from 65% to 90%. T7-siRNA (B), targeted region at 207 nt to 228 nt and double stranded for 21 nt with 2 nt UU 3' overhangs, was the most effective sequence tested for inhibition of VEGF expression in hRPE cells. Compared with nontransfected cells, the mean fluorescence in hRPE cells transfected with T7-sRNAs was significantly less (P < 0.01). siRNA with a single-base mismatch and ssRNA(+) did not show suppressing effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect (5 to 10 pmol). CONCLUSION: T7-siRNA can effectively and specifically suppress VEGF expression in hRPE cells and may be a new way to treat CNV.

[Cloning, expression of soluble VEGFR2 fragment and its effect on tumor angiogenesis].

OBJECTIVE: To study the anti-tumor angiogenesis effect of soluble VEGF receptor fragment by blocking the combination of VEGF and its receptor in vivo and in vitro. METHODS: RT-PCR technique was used to amplify Flk-1/KDR fragment from embryo mouse liver, which was recombinated to expression vector pET-28b(+) and retrovirus vector PLXSN, which was induced to be expressed, purified and identified with EcoR I and Hind III. Mouse endothelial cells were separated, cultured and identified by immunocytochemistrical staining using VIII factor-related antigen antibody. The expressed product was analyzed about its effect on endothelial cell's growth in vitro with MTT method. The retrovirus vector was transfected to tumor cell lines S180 and B16 by liposome method to observe the biological specificity in vitro after gene transfection. RESULTS: 1000 bp size sVEGFR fragment was amplify from E9, E11 embryo mouse liver tissues, which was recombinated to TA clone vector and identified by sequence analysis. This fragment was cloned to expression vector pET-28b(+), the expressed product was purified and identified correctly. The in vitro study showed this expressed product can effectively inhibit endothelial cell(s), growth and proliferation. The fragment was then cloned to retrovirus vector PLXSN and transfected to tumor cell lines S180 and B16 successfully with RT-PCR and SDS-PAGE. The experiments in vivo showed that the weight of tumor smaller, the size decreased significantly, the microvessel density was fewer and Flk1 protein expression were higher in the group of gene transfection than that of control. CONCLUSION: Soluble VEGFR fragment is a kind of effective gene engineer product for anti-tumor angiogenesis gene therapy and the development of anti-tumor drug.

[Effects of high concentrations insulin on VEGF expression in cultured Müller cells].

OBJECTIVE: To investigate the effects of high concentrations of insulin on the expression of vascular endothelial growth factor (VEGF) in cultured rabbit retinal Müller cells. METHODS: Immunocytochemistry, in situ hybridization and the ELISA method were used to study the effects of different concentrations of insulin on the expression of vascular endothelial growth factor (VEGF) in cultured rabbit retinal Müller cells qualitatively and quantitatively. RESULTS: VEGF expression was enhanced obviously by the insulin, which regulated the expression of VEGF at the transcription level. CONCLUSIONS: VEGF expression in cultured Müller cells can be enhanced by high concentrations of insulin. Müller cells play a potential role in the development of neovascularization of diabetic retinopathy.

Diabetic retinopathy: VEGF, bFGF and retinal vascular pathology.

BACKGROUND: Previous research indicated that the development of diabetic retinopathy (DR) is closely related to the excessive expression of growth factors. This paper was to study the relationship of DR with vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and the retinal vascular pathological change. METHODS: Fifty-five Wistar rats, weighing 100 - 200 g, were selected and randomly divided into four groups: control group (no streptozocin injection, n = 10), M1 group (streptozocin induced diabetes for 1 month, n = 15), M3 group (streptozocin induced diabetes for 3 months, n = 15), and M5 group (streptozocin induced diabetes for 5 months, n = 15). In situ hybridization and immunohistochemistry were used to investigate the expressions of bFGF and VEGF on retinal vascular, and retinal vessels were observed by transmission electron microscope. RESULTS: There was no difference in the number of pericytes between M1 and control group (P > 0.05), but the number of pericytes decreased obviously in M3 and M5 groups compared with the control group (P < 0.01, P < 0.001, respectively). Capillary embolization and non-cell capillary were seen in M5 group. Positive expression of VEGF was found in M5 group using in situ hybridization and immunohistochemistry respectively. Positive expression of bFGF could be seen in M3 (78%) and M5 group (89%). Most remarkable changes of vessels were observed in M5 group including fragmental thickness, split of basement membrane, swelling and distortion of endothelial cells. CONCLUSIONS: In retinal vascular of the streptozocin (STZ) rats, there shows the expression of bFGF at the third month and that of VEGF at the fifth month.