Functionalized Collagen/Poly(ethylene glycol) Diacrylate Interpenetrating Network Hydrogel Enhances Beta Pancreatic Cell Sustenance.

Three-dimensional matrices are a new strategy used to tackle type I diabetes, a chronic metabolic disease characterized by the destruction of beta pancreatic cells. Type I collagen is an abundant extracellular matrix (ECM), a component that has been used to support cell growth. However, pure collagen possesses some difficulties, including a low stiffness and strength and a high susceptibility to cell-mediated contraction. Therefore, we developed a collagen hydrogel with a poly (ethylene glycol) diacrylate (PEGDA) interpenetrating network (IPN), functionalized with vascular endothelial growth factor (VEGF) to mimic the pancreatic environment for the sustenance of beta pancreatic cells. We analyzed the physicochemical characteristics of the hydrogels and found that they were successfully synthesized. The mechanical behavior of the hydrogels improved with the addition of VEGF, and the swelling degree and the degradation were stable over time. In addition, it was found that 5 ng/mL VEGF-functionalized collagen/PEGDA IPN hydrogels sustained and enhanced the viability, proliferation, respiratory capacity, and functionality of beta pancreatic cells. Hence, this is a potential candidate for future preclinical evaluation, which may be favorable for diabetes treatment.

Gene Expression Profiling of Markers of Inflammation, Angiogenesis, Coagulation and Fibrinolysis in Patients with Coronary Artery Disease with Very High Lipoprotein(a) Levels Treated with PCSK9 Inhibitors.

Besides lipids, inflammation, angiogenesis, coagulation and fibrinolysis play very important roles in coronary artery disease (CAD). We measured gene expression of the inflammatory markers interleukin (IL)-1ß (IL1B) and interferon (IFN)-γ (IFNG), vascular endothelial growth factor-A (VEGF-A) (VEGFA), and coagulation and fibrinolysis markers tissue factor (TF) (F3) and plasminogen activator inhibitor-1 (PAI-1) (SERPINE) in healthy controls and CAD patients with high lipoprotein(a) (Lp(a)). The aim of our study was to identify, first, if there is a difference in these markers between controls and patients; secondly, if these markers are associated with lipids; and third, what the influence of proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors is on these markers. We included 124 subjects, 27 controls and 97 patients with CAD (30 in placebo and 67 in the PCSK9 group). Blood samples were collected for lipid and gene measurement. The results showed higher expression of IL1B (p < 0.0001), VEGFA (p < 0.0001), and F3 (p = 0.018) in controls in comparison with patients. Significant correlations were observed between IL1B and lipids. Treatment with PCSK9 inhibitors increased VEGFA (p < 0.0001) and F3 (p = 0.001), and decreased SERPINE (p = 0.043). The results of our study underpin the importance of IL-1ß, VEGF-A and TF in CAD as well as the effect of PCSK9 treatment on these markers.

Effect of Ethyl Acetate Extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on Intestinal Flora in ApoE-/- Mice with Atherosclerosis / 中国实验方剂学杂志

ObjectiveTo observe the effect of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on high-fat diet-induced apolipoprotein E gene knockout （ApoE-/-） mice， and explore its mechanism of treating atherosclerosis by regulating intestinal flora. MethodThirty-two 8-week-old male ApoE-/- mice were randomly divided into model group， rosuvastatin group （10 mg·kg-1）， high-， low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis （75， 25 mg·kg-1）， with 8 mice in each group. Eight C57BL/6 mice were used as blank group. After 8 weeks of continuous administration， blood was taken to determine the blood lipid level. Enzyme-linked immunosorbent assay （ELISA） was used to detect the contents of related indexes in serum of mice. Hematoxylin-eosin （HE） staining was used to observe the formation of aortic plaque in mice. Cecal contents were collected and 16S rRNA amplicon sequencing was used to detect intestinal flora. ResultCompared with the blank group， the plaque area of the model group was significantly increased with inflammatory infiltration， the contents of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， inflammatory factors and inducible nitric oxide synthase （iNOS） were increased， while the content of high-density lipoprotein cholesterol （HDL-C） was decreased. Compared with the model group， rosuvastatin group and high- and low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could improve the deposition of aortic plaque， reduce the contents of TG， TC， LDL-C， inflammatory factors and iNOS， and increase the content of HDL-C. Compared with the blank group， the relative abundances of Firmicutes and Proteobacteria in the model group increased， while the relative abundance of Bacteroidetes decreased. Alpha and Beta diversity analysis showed that samples of each group could be significantly isolated， and the total number and abundance of intestinal flora species in the model group were low. Compared with the model group， ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could increase the relative abundance of beneficial bacteria and decrease the relative abundance of pathogenic bacteria. ConclusionEthyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis was mainly composed of flavonoids， which can treat atherosclerosis by regulating the intestinal flora and improve the pathological changes in the aorta of ApoE-/- mice induced by high-fat diet. The mechanism may be related to its ability to reduce the level of inflammatory factors， improve antioxidant capacity and repair the disorder of intestinal flora structure.

Correlation between aneuploidy pregnancy and the concentration of various hormones and vascular endothelial factor in follicular fluid as well as the number of acquired oocytes.

Objective To investigate the correlation between aneuploidy pregnancy and the concentration of various hormones and vascular endothelial factor in follicular fluid as well as the number of acquired oocytes and to provide a scientific basis for improving ovulation induction programs. Methods In total, we collected 277 follicular fluid specimens from patients undergoing in vitro fertilization (IVF) treatment in our hospital. Eighteen cases of aneuploidy embryos were identified. The follicular fluid of these aneuploidy embryos was used for the study. According to the case and control 1:5 paired design, we selected five age-matched controls with healthy births following IVF for each aneuploidy case. Concentrations of anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrogen (E2) and vascular endothelial growth factor (VEGF) in the follicular fluid were measured. Conditional logistic regression was used to analyze the relationship between aneuploidy pregnancy and the concentrations of various hormones and VEGF in the follicular fluid as well as the number of acquired oocytes. Results Multivariate conditional logistic regression showed that of all the factors analyzed, only FSH [odds ratio (OR) = 1.300, 95% confidence interval (CI), 1.091-1.548, P = 0.003] level in the follicular fluid and the number of acquired oocytes (OR = 1.179, 95% CI, 1.070-1.299, P = 0.001) were closely related to aneuploidy pregnancy. No other factors were found to be associated with aneuploidy pregnancy. Conclusion FSH concentrations in the follicular fluid are risk factors for aneuploidy pregnancies. The higher the number of eggs, the higher the risk of aneuploidy. These findings may help improve ovulation induction programs.

[Efficacy of compound Dendrobium on PI3K/AKT/eNOS signaling pathway in hypertensive rats induced by "dietary disorders"].

To observe the efficacy of compound Dendrobium on Sprague Dawley rats (SD) hypertension model induced by "dietary disorders" and its relevant mechanism, totally 50 SD rats were fed with high-sugar, high-fat diet and alcohol for four weeks. According to the blood pressure after modeling, the rats were divided into model group, valsartan group (8 mg·kg⁻¹), low, medium and high-dose Dendrobium candidum compound groups (1.65, 3.30, 5.00 g·kg⁻¹), with 10 rats in each group, and the other 10 SD rats were also taken as the normal group. After four weeks of treatment, blood pressure was measured. Orbital blood was collected for the determination of serum cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL), calculation of atherosclerosis index (AI). Nitric acid reductase method was used to detect serum nitric oxide (NO); the levels of serum endothelin-1 (ET-1) and intercellular adhesion molecule-1 (ICAM-1) were measured by ELISA. The rats were put to death after the last administration, and the protein expressions of PI3K/AKT/eNOS in thoracic aorta of rats in each group were detected by Western blot. The aorta was separated and stained with hematoxylin-eosin (HE) to observe the changes in the endothelium and blood vessels in the thoracic aorta. Masson staining was used to observe the formation of aortic collagen. The expressions of nitric oxide synthase (eNOS) and ICAM-1 in aortic endothelial cells were observed by immunohistochemistry. In contrast, the results show D. candidum compound can significantly reduce the blood pressure in hypertensive rats, increase HDL-c, and reduce AI, while increasing serum NO content, decreasing ET-1 and ICAM-1 levels and promoting PI3K/AKT/NOS protein expressions. The lesion degree of the D. candidum compound group was reduced, and the collagen deposition was significantly reduced. Meanwhile, D. candidum compound can significantly increase the expression of eNOS, and reduce the formation of ICAM-1.Therefore, D. candidum compound has an obvious antihypertensive effect on hypertensive rats, which may be related to the increase in PI3K/AKT/eNOS signaling pathways and NO generation, the inhibition of the secretion of ICAM-1 and ET-1, the protection of the vascular endothelium and the improvement of aortic disease.

Research on AT-Ⅲ, PAI and t-PA in patients with different stages of diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To explore the relationship between the different stages of diabetic retinopathy and the related factors of vascular endothelial function,and to provide a theoretical basis for improving the function of vascular endothelium to find a way to delay or even inhibit the occurrence or progression of DR.METHODS:We collected during March 2015 to December 2015 in Department of Ophthalmology and endocrinology in our hospital,178 cases of type 2 diabetes mellitus patients and 62 cases of blood specimen in health control group.According to the results of fundus fluorescence angiography (FFA),they were divided into four groups,diabetes patients without retinopathy,diabetes patients with non proliferative diabetic retinopathy (NPDR),diabetes patients with proliferative diabetic retinopathy (PDR) and healthy control group.We detected blood samples of antithrombin Ⅲ (AT-Ⅲ),fibrinolytic enzyme activation inhibitor (PAI),the original organization type fibrinolytic enzyme activator (t-PA) index and the correlation of diabetic retinopathy in installment.RESULTS:This study showed that AT-Ⅲ was significantly different among the four groups (F=5.986,P＜ 0.01);PAI was significantly different among the patients without DR,patients with NPDR and patients with PDR (F=7.434,P＜0.01);t-PA was not significantly different among the four groups (F=2.556,P＞ 0.05);there were relations between the different stages of diabetic retinopathy and AT-Ⅲ,PAI.CONCLUSION:The degree of diabetic retinopathy has a close relationship with the content of antithrombin Ⅲ and plasminogen activator inhibitor,and it is closely related to the function of vascular endothelium.

Efficacy of compound Dendrobium on PI3K/AKT/eNOS signaling pathway in hypertensive rats induced by "dietary disorders" / 中国中药杂志

To observe the efficacy of compound Dendrobium on Sprague Dawley rats (SD) hypertension model induced by "dietary disorders" and its relevant mechanism, totally 50 SD rats were fed with high-sugar, high-fat diet and alcohol for four weeks. According to the blood pressure after modeling, the rats were divided into model group, valsartan group (8 mg·kg⁻¹), low, medium and high-dose Dendrobium candidum compound groups (1.65, 3.30, 5.00 g·kg⁻¹), with 10 rats in each group, and the other 10 SD rats were also taken as the normal group. After four weeks of treatment, blood pressure was measured. Orbital blood was collected for the determination of serum cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL), calculation of atherosclerosis index (AI). Nitric acid reductase method was used to detect serum nitric oxide (NO); the levels of serum endothelin-1 (ET-1) and intercellular adhesion molecule-1 (ICAM-1) were measured by ELISA. The rats were put to death after the last administration, and the protein expressions of PI3K/AKT/eNOS in thoracic aorta of rats in each group were detected by Western blot. The aorta was separated and stained with hematoxylin-eosin (HE) to observe the changes in the endothelium and blood vessels in the thoracic aorta. Masson staining was used to observe the formation of aortic collagen. The expressions of nitric oxide synthase (eNOS) and ICAM-1 in aortic endothelial cells were observed by immunohistochemistry. In contrast, the results show D. candidum compound can significantly reduce the blood pressure in hypertensive rats, increase HDL-c, and reduce AI, while increasing serum NO content, decreasing ET-1 and ICAM-1 levels and promoting PI3K/AKT/NOS protein expressions. The lesion degree of the D. candidum compound group was reduced, and the collagen deposition was significantly reduced. Meanwhile, D. candidum compound can significantly increase the expression of eNOS, and reduce the formation of ICAM-1.Therefore, D. candidum compound has an obvious antihypertensive effect on hypertensive rats, which may be related to the increase in PI3K/AKT/eNOS signaling pathways and NO generation, the inhibition of the secretion of ICAM-1 and ET-1, the protection of the vascular endothelium and the improvement of aortic disease.

One-year outcomes of ziv-aflibercept for macular edema in central retinal vein occlusion.

PURPOSE: To report the 12-month efficacy and safety outcomes of intravitreal ziv-aflibercept in macular edema secondary to central retinal vein occlusion (CRVO). METHODS: Interventional case series documenting 12-month outcomes of intravitreal ziv-aflibercept (1.25 mg in 0.05 mL) in 6 patients with treatment-naive macular edema secondary to CRVO. All patients had comprehensive ophthalmic examination, spectral domain optical coherence tomography at baseline and all follow-up visits, and fluorescein. Retreatment decisions were based on recurrence or persistence of intraretinal or subretinal fluid, deterioration in visual acuity (VA), increase in central subfield thickness (CST) by ≥ 50 µm from the previous visit, or lowest recorded CST. RESULTS: Participants had (2 males, 4 females) an average age of 53.5 years. From baseline to 12 months, the mean logMAR VA improved from 0.86 (Snellen ≈ 20/145) to 0.33 (Snellen ≈ 20/40), central macular thickness decreased from 519 µm to 255 µm, and total macular volume decreased from 14.7 mm3 to 7.1 mm3. No eyes had uveitis, cataract progression, intraocular pressure (IOP) elevations, or systemic adverse events. CONCLUSIONS AND IMPORTANCE: Ziv-aflibercept achieves favorable intermediate-term functional and structural outcomes in macular edema secondary to CRVO. No safety concerns were raised. Low-cost ziv-aflibercept may thus be useful for CRVO in resource-poor countries. Further prospective studies in larger cohorts are needed further establish the efficacy and safety of this agent.

Biodegradable Carriers for Delivery of VEGF Plasmid DNA for the Treatment of Critical Limb Ischemia.

The safe and efficient delivery of therapeutic nucleic acid is a prerequisite for an effective DNA therapy. In this study, we condensed the low molecular weight polyethylenimine (PEI, 1.8k Da) with 2,6-pyridinedicarboxaldehyde (PDA), both of which are degradable in vivo, to synthesize a biodegradable polycationic material (PDAPEI) to deliver vascular endothelial growth factor (VEGF) plasmid DNA (pDNA). Particle size and zeta potential of this novel degradable PEI derivatives-pDNA nanoparticle were investigated and in vitro cytotoxicity was estimated on human umbilical vein endothelial cells (HUVECs). Using pDNA-encoding VEGF-A and green fluorescence protein (GFP), we also checked transfection efficiency of the vector (PDAPEI) and found its excellent performance at 40 w/w ratio. We successfully established peripheral ischemia animal model on C57/BL6J mice to evaluate the therapeutic effect of PDAPEI/pVEGF-A polyplex system on ischemic disease and a conclusion was made that PDAPEI is a promising gene vector in the treatment of peripheral ischemic artery disease (PAD).

Vascular endothelial factor C and E-cadherin relationship with the invasion and metastasis of human gastric cancer / 中国综合临床

Objective To investigate the expression of vascular endothelial factor C(VEGF-C) and E-cadherin in gastric cancer and explore the relationship between expression of VEGF-C and E-cadherin and the pathogenesis of gastric cancer.Methods Real-time quantitative reverse ranscriptase-polymerase chain rection was applied on 40 cases of gastric cancer and corresponding adjacent noncancerous tissues,in order to detect mRNA expression of VEGF-C and E-cadherin gene.VEGF-C and E-cadherin protein expression in gastric cancer and adjacent normal gastric mucosa were detected by immunohistochemistry.Statistical analysis was carried out to analyze the correlation among VEGF-C,E-cadherin and protein expression with various clinical parameters in these gastric cancer patients.Results The expression of VEGF-C protein in 40 cases of gastric cancer's cancer tissues was 0.461±0.012,significantly higher than that in the adjacent normal tissues(0.036+0.023;t=1.101,P＜0.05),but E-cadherin expression was significantly lower than that of the adjacent normal tissues (0.079±0.002 vs.0.321±0.005;t=1.844,P＜0.05).There was correlation between VEGF-C mRNA expression with histological grading,TNM staging,lymph node and distant metastasis (t=-1.621,-1.474,-2.378,-1.966,P＜0.05).There was correlation between E-cadherin mRNA expression with tumor size,histological grading,TNM stage,there was a significant difference (t=1.875,1.673,1.544,P＜0.05).VEGF-C and E-cadherin protein expression was negatively correlated(r=-0.688,P＜0.05).Conclusion Up-regulated of VEGF-C gene and decreased expression of E-cadherin may play an important role in the carcinogenesis of gastric cancer

Combination of growth factor treatment and scaffold deposition following traumatic brain injury has only a temporary effect on regeneration.

The recovery after traumatic brain injury (TBI) is hampered by the poor regenerative capacity of the brain. Today there is no treatment available that effectively restores lost brain tissue, but much research is focused on the stimulation of endogenous neural stem cells to viably and functionally repopulate the injured parenchyma. It is crucial that the therapies have a proven long-term effect on both regeneration and functional recovery to be clinically interesting. Here we have studied the induction of stem cell activation in rats at three weeks and six weeks after inducing TBI using controlled cortical impact model at a severe setting. We combined intracerebroventricular growth factor and scaffold treatment in order to accomplish an optimal effect on the stem cell regeneration. Immediately after TBI epidermal growth factor infusion with osmotic minipumps was started and continued for seven days. The pumps were removed and an extracellular matrix scaffold containing vascular endothelial growth factor was deposited into the cortical cavity. Three weeks after injury there was a positive effect of the treatment with a significant increase in neuronal and astrocytic regeneration. However, after six weeks there was no difference in the number of newly generated neurons and astrocytes in treated or untreated rats. Evaluation of tissue loss and spatial learning in the Morris water maze corroborated that the treatment had no effect at the later time point. Our results highlight the importance of long-term studies to ensure that a promising effect on tissue regeneration and functional outcome is not only temporary.

Protection of Intrahepatic Injection of Liposome-mediated VEGF Plasmid against Ischemia-reperfusion Liver Injury / 华中科技大学学报(医学版)

Objective To explore the effect of intrahepatic injection of liposome-mediated VEGF plasmid on ischemia-reper-fusion liver injury and its mechanism. Methods Rabbits were randomly divided into normal group, ischemia-reperfusion group and recombinant VEGF therapy group( liposome-mediated transfer of VEGF plasmid into liver via portal vein 20 min before ischemia of liver). The model of liver ischemia-reperfusion injury was established. Liver function and the activity of SOD.XO in blood were determined at the 0,2nd,6th,12th,and 24th h after operation. RT-PCR technique was applied to detect the expression level of Fas mRNA in liver tissues of every group,and flow cytometry was used to measure cell apoptosis rate at the 6th h after operation. At the 24th h after operation,all rabbits were killed and liver tissues of ischemia were taken to make pathological sections for observing the morphology and microstructure under the light microscopy and electron microscopy. ResuJts The level of ALT in recombinant VEGF therapy group was markedly reduced as compared with ischemia-reperfusion group at the 6th,12th,and 24th h after operation( P<0. 05). The activity of SOD in recombinant VEGF therapy group was significantly higher than in ischemia-reperfusion group at the 6th, 12th,and 24th h after operation. The activity of XO in recombinant VEGF therapy group was significantly lower than that in ischemia-reperfusion group at the 6th,12th,and 24th h after operation(P< 0. 05 or P<0. 01). In addition,there was significant difference in the expression of Fas mRNA and cell apoptosis rate between recombinant VEGF therapy group and ischemia-reperfusion group(P<0. 01). The injury of hepatocytes in recombinant VEGF therapy group was significantly alleviated as compared with that in ischemia-reperfusion group under the light microscopy and e-lectron microscopy. Conclusion Liposome-mediated transfer of VEGF plasmid into liver before ischemia of liver can obviously protect hepatocytes by increasing anti-oxidative ability, decreasing the expression of Fas mRNA, and finally inhibiting hepato-cyte apoptosis.

Effects of exogenous antisense VEGF-C gene on growth of human gastric carcinoma cell line SGC-7901 in vitro / 重庆医科大学学报

Objective:To observe the expression of VEGF-C protein in human gastric cancer cells SGC-7901 those transfected by pCI-neo-antisense VEGF-C and investigate whether antisense VEGF-C gene could inhibit the growth of human gastric carcinoma cell line in vitro.Methods:By the liposome infection protocol,We transfected two different plasmids including pCI-neo and pCI-neo antisense VEGF-C into human gastric cancer cell line SGC-7901 cultured in vitro.Then those resistant clones were selected by G418 and expanded in DMEM culture medium.Then we used Immunohistochemical method and Western-blot to detect target genes and its expression.The cell viability was detected by MTT assay.We took the untransfected group as the control.Results:The immunohistochemical method and Western-blot showed that there were no expression of VEGF-C mRNA and its protein in the group that transfected by plasmid pCI-neo antisense VEGF-C.MTT assay showed that the group of transfected with antisense VEGF-C had a lower cell viability than other groups(P

Expression of paxillin and vascular endothelial growth factor in lung carcinoma and its clinical implications / 中国实用内科杂志

Objective To investigate the expressions of paxillin and vascular endothelial growth factor (VEGF) in lung carcinoma.Method SP immunohistological staining was used to detect the expression of paxillin and VEGF in 79 patients with lung carcinoma.Result 1.The expression rate of PAX and VEGF was 34.1% and 64.6%respectively.2.Significant positive correlation was found between the expression of PAX and lymphoid metastasis (P

The expression of VEGF-D and prognosis of patients with colorectal carcinoma / 中国普通外科杂志

Objective To evaluate the expression of vascular endothelial factor ( VEGF) -D in predicting the prognosis of colorectal carcinoma (CRC). Methods Between Jan 1996 and Jan 1998, 69 patients with CRC undergoing curative surgery were included in this study. Postoperative follow-up included physical examination, serum CEA, and imaging every 3 months in the first and the second year, every 6 months in the third year and once a year thereafter. The expression of VEGF-D protein and microvessel density ( MVD ) in 69 tissues of CRC and 20 normal colorectal tissues was examined by immunohistochemistry (IHC). Results of IHC staining were quantified by Axioplan 2 imaging analysis system. Results VEGF-D protein expression in the cytoplasm was found in all of the CRC tissues and 25% (5/20 ) of normal tissues. The VEGF-D expression was much higher in tumor tissue than in the corresponding normal tissue (P