Neovascularization by Sustained Delivery of G-CSF, EPO and VEGF Using Dextran/PLGA Microspheres.

BACKGROUND: Therapeutic neovascularization has some obstacles, such as it requires more than one proangiogenic factor, and these factors have short half-lives. To overcome these obstacles, combined delivery of granulocyte-colony stimulating factor (G-CSF), erythropoietin (EPO) and vascular endothelial growth factor (VEGF) using protein/dextran/poly (lactic-co-glycolic acid) (PLGA) sustained-release microspheres was proposed to promote neovascularization. METHODS: Dextran microparticles loaded with G-CSF, EPO or VEGF were prepared and encapsulated in PLGA microspheres to obtain protein-dextran-PLGA microspheres. The release behavior of microspheres was studied in vitro. The protein/dextran/PLGA microspheres were injected into the ischemic hindlimbs of rats. Neovascularization in ischemic muscle was measured. RESULTS: Microspheres released G-CSF, EPO and VEGF in vitro for more than 4 weeks. Combined therapy with VEGF, EPO and G-CSF promoted the expression of B-cell lymphoma-2 and stromal cell-derived factor 1, cellular proliferation and the incorporation of C-X-C chemokine receptor 4 positive cells. Capillary density and smooth muscle α-actin+ vessel density were higher in the combined treatment of VEGF, EPO and G-CSF than in the single factor treatment. CONCLUSIONS: The combined and sustained delivery of VEGF, EPO and G-CSF using dextran-PLGA microspheres had a more significant neovascularization effect than monotherapy with each factor alone. This combined therapy might be a promising treatment for ischemic vascular diseases.

Increase of wall shear stress caused by arteriovenous fistula reduces neointimal hyperplasia after stent implantation in healthy arteries.

BACKGROUND AND OBJECTIVES: Wall shear stress plays a critical role in neointimal hyperplasia after stent implantation. It has been found that there is an inverse relation between wall shear stress and neointimal hyperplasia. This study hypothesized that the increase of arterial wall shear stress caused by arteriovenous fistula could reduce neointimal hyperplasia after stents implantation. METHODS AND RESULTS: Thirty-six male rabbits were randomly divided into three groups: STENT, rabbits received stent implantation into right common carotid artery; STENT/arteriovenous fistula, rabbits received stent implantation into right common carotid artery and carotid-jugular arteriovenous fistula; Control, rabbits received no treatment. After 21 days, stented common carotid artery specimens were harvested for histological staining and protein expression analysis. In STENT group, wall shear stress maintained at a low level from 43.2 to 48.9% of baseline. In STENT/arteriovenous fistula group, wall shear stress gradually increased to 86% over baseline. There was a more significant neointimal hyperplasia in group STENT compared with the STENT/arteriovenous fistula group (neointima area: 0.87 mm2 versus 0.19 mm2; neointima-to-media area ratio: 1.13 versus 0.18). Western blot analysis demonstrated that the protein level of endothelial nitric oxide synthase in STENT group was significantly lower than that in STENT/arteriovenous fistula group, but the protein levels of proliferating cell nuclear antigen, vascular cell adhesion molecule 1, phospho-p38 mitogen-activated protein kinase (Pp38), and phospho-c-Jun N-terminal kinase in STENT group were significantly higher than that in the STENT group. CONCLUSION: High wall shear stress caused by arteriovenous fistula as associated with the induction in neointimal hyperplasia after stent implantation. The underlying mechanisms may be related to modulating the expression and activation of endothelial nitric oxide synthase, vascular cell adhesion molecule 1, p38, and c-Jun N-terminal kinase.

Sustained delivery of vascular endothelial growth factor using a dextran/poly(lactic-co-glycolic acid)-combined microsphere system for therapeutic neovascularization.

We hypothesize that the controlled delivery of vascular endothelial growth factor (VEGF) using a novel protein sustained-release system based on the combination of protein-loaded dextran microparticles and PLGA microspheres could be useful to achieve mature vessel formation in a rat hind-limb ischemic model. VEGF-loaded dextran microparticles were fabricated and then encapsulated into poly(lactic-co-glycolic acid) (PLGA) microspheres to prepare VEGF-dextran-PLGA microspheres. The release behavior and bioactivity in promoting endothelial cell proliferation of VEGF from PLGA microspheres were monitored in vitro. VEGF-dextran-PLGA microsphere-loaded fibrin gel was injected into an ischemic rat model, and neovascularization at the ischemic site was evaluated. The release of VEGF from PLGA microspheres was in a sustained manner for more than 1 month in vitro with low level of initial burst release. The released VEGF enhanced the proliferation of endothelial cells in vitro, and significantly promoted the capillaries and smooth muscle α-actin positive vessels formation in vivo. The retained bioactivity of VEGF released from VEGF-dextran-PLGA microspheres potentiated the angiogenic efficacy of VEGF. This sustained-release system may be a promising vehicle for delivery of multiple angiogenic factors for therapeutic neovascularization.

[Study on concomitant surgical correction of pelvic organ prolapse and TVT-O for treatment of stress urinary incontinence].

OBJECTIVE: To investigate the necessity, safety and efficacy of transobturator tension-free vaginal tape (TVT-O) for treatment of stress urinary incontinence (SUI) during transvaginal corrective operation of pelvic organ prolapse (POP). METHODS: From Jan. 2005 to Dec. 2010, 92 patients undergoing transvaginal pelvic reconstruction surgery for correction of POP concomitant TVT-O for treatment of SUI in Department of Obstetrics and Gynecology affiliated to Beijing Chaoyang Hospital as concomitant surgery group were enrolled in this retrospective study matched with 90 patients with mild SUI without SUI surgery as non-concomitant surgery group and 120 patients without SUI as control group.Variable clinical index, clinical efficacy and complications were compared among those three groups. RESULTS: Compared with those in the other two groups, the mean age [(62 ± 11) years] was lower (P = 0.007,0.038), the operation time only slightly increased (12.8 min and 12.9 min respectively) significantly in concomitant TVT-O group. The bleeding loss and the length of staying hospital after operation all exhibited no significant differences within three groups (P > 0.05). The effective rate for SUI was 96.7% (89/92) in concomitant TVT-O group, corrective operation of POP was ineffective for 74.4% (67/90) SUI, 9.2% (11/120) patients presented new SUI in the patients without SUI preoperatively. CONCLUSIONS: TVT-O is a simple, safe and effective method in the treatment of SUI, which is more suitable for performing simultaneously during the corrective operation of POP.Efficacy of SUI correction was limited in those patients undergoing only pelvic reconstructive surgery. However, a preventive anti-incontinence procedure is not recommended because of the lower incidence in POP patients without SUI preoperatively.

The correlation and role analysis of SLC30A1 and SLC30A10 in cervical carcinoma.

Background: SLC30 family genes, also known as ZnT family genes, can keep cellular zinc levels within a physiological range by exporting zinc to extracellular space or by isolating zinc in the specific regions of cytoplasm when cellular zinc concentrations are elevated in human cells. There are growing evidences that dysregulated expression of SLC30 family genes can potentially influence tumorigenesis. However, the expression and prognostic value of SLC30 family genes in cervical carcinoma are poorly characterized. Methods: In this study, we used many tools such as UALCAN, Kaplan-Meier Plotter, cBioPortal, LinkedOmics, FunRich, Metascape, GeneMANIA, Open targets and TISIDB to perform bioinformatics analysis of SLC30 family genes in cervical carcinoma. Results: We found that the expression of SLC30A1/7/10 was significantly higher in cervical carcinoma than that in normal matched tissues, while SLC30A2/8 mRNA levels were decreased compared to normal tissues. For tumor stages, SLC30A1, SLC30A7 and SLC30A10 groups significantly varied. And a high expression of SLC30A1, SLC30A6, SLC30A8 and SLC30A10 was associated with worse overall survival in cervical carcinoma patients. Besides, we found that SLC30A1/10 may have a potential regulatory role in immune infiltration in cervical carcinoma. In addition, the results showed that the high expression of SLC30A1 was resistant to 79 drugs or small molecules; Two drugs (Neopeltolide and Tozasertib) can inhibit the high expression of SLC30A10 in cancers. Conclusion: SLC30A1 and SLC30A10 can be recognized as potential diagnostic indicators and therapeutic targets in cervical carcinoma.

Transthyretin increases migration and invasion of rat placental trophoblast cells.

Preeclampsia (PE) is a hypertensive disorder of pregnancy. Early diagnosis of PE is currently contingent on regular prenatal physical examinations and may be facilitated by identification of novel diagnostic markers. Transthyretin (TTR), also known as prealbumin, is primarily responsible for maintaining the normal levels of thyroxine and retinol binding protein. The expression of TTR is lower in patients with severe PE as compared with healthy controls. Here, we examined the suitability of TTR as a diagnostic marker in pregnant hypertensive rats. N'-nitro-l-arginine-methylesterhydrochloride (l-NAME) was used to generate a rat model of hypertension during pregnancy. Rat placental trophoblast cells were divided into control and TTR groups for in vitro experiments. Systolic blood pressure, diastolic blood pressure, mean blood pressure and urinary protein of hypertensive pregnant rats were higher than those of healthy pregnant rats, but these effects could be reversed by TTR treatment. There were no significant changes in blood pressure and urinary protein in healthy pregnant rats before or after TTR treatment. TTR levels in the serum and placental tissues of pregnant hypertensive rats were significantly reduced compared with those of healthy pregnant rats. Changes in placental and fetal weights in the hypertensive model could also be rescued by TTR treatment. TTR treatment significantly increased the level of matrix metalloproteinase-2/9 in hypertensive rats. Finally, in vivo and in vitro experiments demonstrated that TTR effectively increased the migration and invasion of rat placental trophoblast cells, as well as matrix metalloproteinase-2/9 levels in these cells. In conclusion, our data from a rat model suggest that TTR may have potential as a novel marker for PE diagnosis.

[Prospective investigation of postoperative lower extremity deep venous thrombosis in gynecological procedures].

OBJECTIVE: To investigate the morbidity, relevant factors and diagnostic methods of lower extremity deep venous thrombosis (LEDVT) after gynecological pelvic surgeries. METHODS: A prospective study was carried out investigating the morbidity, characteristics, risk factors, the coagulation function changes and diagnostic methods of LEDVT post-gynecological pelvic operations in 141 cases. Platelet, prothrombin time, activated partial thromboplastin time, fibrinogen, thrombin time, D-dimer, antithrombin-III, tissue plasminogen activator, plasminogen activator inhibitor were measured within 1 week before surgeries, 48 hours and 5 - 7 days post-surgeries respectively. Color Doppler flow imaging was performed simultaneously. RESULTS: (1) Twenty-two cases were found with LEDVT, the morbidity of LEDVT after gynecological procedures was 15.6%. It occurred 2 - 7 days post operation. (2) The risk factors were bedfasting, coagulant prescription post-operation, cardiovascular disease, ageing, pelvic lymphadenectomy, general anesthesia, and malignancy. Multivariate regression showed only age and coagulant prescription post-operation were independent factors. (3) The level of D-dimer was (0.9 +/- 0.6) mg/L in LEDVT group, significantly higher than that of non-LEDVT group [(0.5 +/- 0.4) mg/L, P < 0.01]. The positive predictive value and negative predictive value of D-dimer were 30.8% and 98.6% respectively. (4) Among 22 cases who were diagnosed with LEDVT by compression duplex ultrasound, only 4 cases performed lower extremity venography, and the diagnosis was confirmed. CONCLUSIONS: The morbidity of postoperative lower extremity deep venous thrombosis in gynecological procedures was high. The combination of D-dimer and compression duplex ultrasound examination was applicable to the diagnosis of LEDVT. The optional duration of the examination was 2 - 7 days post operations.

Phosphoproteomics Analysis of Endometrium in Women with or without Endometriosis.

BACKGROUND: The molecular mechanisms underlying the endometriosis are still not completely understood. In order to test the hypothesis that the approaches in phosphoproteomics might contribute to the identification of key biomarkers to assess disease pathogenesis and drug targets, we carried out a phosphoproteomics analysis of human endometrium. METHODS: A large-scale differential phosphoproteome analysis, using peptide enrichment of titanium dioxide purify and sequential elution from immobilized metal affinity chromatography with linear trap quadrupole-tandem mass spectrometry, was performed in endometrium tissues from 8 women with or without endometriosis. RESULTS: The phosphorylation profiling of endometrium from endometriosis patients had been obtained, and found that identified 516 proteins were modified at phosphorylation level during endometriosis. Gene ontology annotation analysis showed that these proteins were enriched in cellular processes of binding and catalytic activity. Further pathway analysis showed that ribosome pathway and focal adhesion pathway were the top two pathways, which might be deregulated during the development of endometriosis. CONCLUSIONS: That large-scale phosphoproteome quantification has been successfully identified in endometrium tissues of women with or without endometriosis will provide new insights to understand the molecular mechanisms of the development of endometriosis.

Two mechanical methods for thromboembolism prophylaxis after gynaecological pelvic surgery: a prospective, randomised study.

BACKGROUND: Venous thromboembolism is known to be an important social and health care problem because of its high incidence among patients who undergo surgery. Studies on the mechanical prophylaxis of thromboembolism after gynaecological pelvic surgery are few. The aim of our study was to evaluate the effect of mechanical thromboembolism prophylaxis after gynaecological pelvic surgery using a combination of graduated compression stockings (GCS) and intermittent pneumatic compression (IPC) or GCS alone. METHODS: The study was performed on 108 patients who were randomly assigned to two groups. The first group received GCS before the operation and IPC during the operation (IPC + GCS group). The second group received GCS before the operation (GCS group). To analyze the effect of the preventive measures and the laboratory examination on the incidence of thrombosis and to compare the safety of these measures, the incidence of adverse reactions was assessed. RESULTS: The morbidity associated with DVT was 4.8% (5/104) in the IPC + GCS group and 12.5% (14/112) in the GCS group. There were significant statistical differences between the two groups. There were no adverse effects in either group. CONCLUSIONS: The therapeutic combination of GCS and IPC was more effective than GCS alone for thrombosis prevention in high-risk patients undergoing gynaecological pelvic surgery, and there were no adverse effects in either group.