Novel nano-microspheres containing chitosan, hyaluronic acid, and chondroitin sulfate deliver growth and differentiation factor-5 plasmid for osteoarthritis gene therapy / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

OBJECTIVE@#To construct a novel non-viral vector loaded with growth and differentiation factor-5 (GDF-5) plasmid using chitosan, hyaluronic acid, and chondroitin sulfate for osteoarthritis (OA) gene therapy.@*METHODS@#Nano-microspheres (NMPs) were prepared by mixing chitosan, hyaluronic acid, and chondroitin sulfate. GDF-5 plasmid was encapsulated in the NMPs through electrostatic adsorption. The basic characteristics of the NMPs were observed, and then they were co-cultured with chondrocytes to observe their effects on extracellular matrix (ECM) protein expression. Finally, NMPs loaded with GDF-5 were injected into the articular cavities of rabbits to observe their therapeutic effects on OA in vivo.@*RESULTS@#NMPs exhibited good physicochemical properties and low cytotoxicity. Their average diameter was (0.61±0.20) μm, and encapsulation efficiency was (38.19±0.36)%. According to Cell Counting Kit-8 (CCK-8) assay, relative cell viability was 75%-99% when the total weight of NMPs was less than 560 μg. Transfection efficiency was (62.0±2.1)% in a liposome group, and (60.0±1.8)% in the NMP group. There was no significant difference between the two groups (P>0.05). Immunohistochemical staining results suggested that NMPs can successfully transfect chondrocytes and stimulate ECM protein expression in vitro. Compared with the control groups, the NMP group significantly promoted the expression of chondrocyte ECM in vivo (P<0.05), as shown by analysis of the biochemical composition of chondrocyte ECM. When NMPs were injected into OA model rabbits, the expression of ECM proteins in chondrocytes was significantly promoted and the progression of OA was slowed down.@*CONCLUSIONS@#Based on these data, we think that these NMPs with excellent physicochemical and biological properties could be promising non-viral vectors for OA gene therapy.

Expression of GDF10 in spinal cord in a rat model of neuropathic pain / 中国病理生理杂志

AIM: To observe the expression change of growth and differentiation factor 10 ( GDF10 ) in the spinal cord of the rats with neuropathic pain .METHODS:Male SD rats (n=60) were used.The neuropathic pain was induced by ligation of left L 5 spinal nerves of the animals .The paw withdrawal threshold was detected 1 d before surgery , and 0 d, 1 d, 3 d, 10 d and 21 d after surgery.The changes of GDF10 in the dorsal horn of L5 spinal cord were detected by immunofluorescence staining and Western blot .RESULTS:The paw withdrawal threshold of the rats with spinal nerve ligation was decreased from 1 d after surgery until 3 d with obvious difference compared with the na ve rats ( P<0.05 ) , continuously decreased until 10 d, and then stabilized at 21 d.The GDF10 was located in the cytoplasm of the neurons in the dorsal horn of L5 spinal cord detected by immunofluorescence staining .The expression of GDF10 in L5 dorsal horn de-tected by immunofluorescence staining was reduced after surgery , significantly decreased from 10 d ( P<0.05) until more than 21 d after surgery in spinal nerve ligation group compared with na ve group.GDF10 in L5 spinal cord detected at 10 d after surgery by Western blot was significantly down-regulated in spinal nerve ligation group compared with na ve group (P<0.05).CONCLUSION:Spinal nerve ligation induces the decrease in GDF 10 expression in spinal dorsal horn .The down-regulation of GDF10 may contribute to the regulation of hyperpathia caused by mechanical stimulation after the injury of spinal nerve .

DNA release and uptake associated with the development of pleomorphic cells in mammalian skin autotransplants / Liberación y absorción de ADN asociada con el desarrollo de células pleomórficas en autotrasplantes de piel en mamíferos

OBJECTIVE: Although several in vitro studies have demonstrated active release of DNA by living cells, there is still doubt. There are no such in vivo studies (1). The following experiment is an in vivo study to determine whether DNA release and uptake by cells and tissues occur and can be related to normal growth and differentiation, abnormal growth and cancer. METHODS: Epidermal and full-thickness ear-skin grafts were separately autotransplanted into two groups of mice. In a second group, host mice were labelled with tritiated thymidine and autografted separately, with unlabelled epidermal and full-thickness ear-skin grafts. Animals were sacrificed regularly in both cases. RESULTS: Full thickness grafts revealed cysts in 15 out of 16 grafts, with well-differentiated squamous epidermis, DNA labelling ofdermal fibroblasts and no DNA labelling ofepidermal cells. Epidermal grafts revealed cysts in six out of 20 grafts, with epidermal cells variable in shape and arrangement; some appeared normal but others were two to four times larger, forming solid nests ofcells. In some grafts, there were spindle-shaped pleomorphic cells loosely interconnected. DNA labelling was ob served in occasional epidermal cell. Two lung adenocarcinomas were found. CONCLUSION: These results suggest active release of DNA by host cells and DNA uptake by grafted cells. This phenomenon and the differential uptake of DNA labelling ofepidermal and dermal cells in the epidermal and full-thickness grafts suggest an association with abnormal, even pleomorphic epidermal cell behaviour due to the interference of dermal/epidermal interacting factors.

OBJETIVO: Aunque varios estudios in vitro han demostrado la liberación activa de DNA por las células vivas, todavía persisten las dudas. No existen tales estudios in vitro (1). El siguiente experimento constituye un estudio in vitro para determinar si hay liberación y absorción de ADN por parte de las células y los tejidos, y si estos procesos guardan relación con el crecimiento normal y la diferenciación, así como con el crecimiento anormal y el cáncer. MÉTODOS: Injertos de piel de la oreja, tanto de espesor total como epidérmicos fueron auto trasplantados por separado a dos grupos de ratones. En el segundo grupo, ratones huéspedes fueron etiquetados con timidina tritiada, y autoinjertados, por separado, con injertos de piel de la oreja no etiquetados, tanto de espesor total como epidérmicos. En ambos casos, fue necesario sacrificar animales de manera regular. RESULTADOS: Los injertos de espesor total revelaron quistes en 15 de cada 16 injertos, con epidermis escamosa bien diferenciada, etiquetado ADN de fibroblastos dérmicos, y no etiquetado ADN de células epidérmicas. Los injertos epidérmicos revelaron quistes en seis de 20 injertos, siendo las células epidérmicas variables en forma y ordenamiento. Algunas parecían normales, pero otras eran de dos a cuatro veces mayores, y formaban anidamientos celulares sólidos. En algunos de los injertos, se presentaron células pleomórficas en forma de huso, interconectadas con laxitud. Se observó etiquetado de DNA en células epidérmicas ocasionalmente. Se hallaron dos adenocarcinomas pulmonares. CONCLUSIÓN: Estos resultados sugieren la liberación activa de ADN por las células huéspedes y la absorción de ADN por las células injertadas. Este fenómeno y la absorción diferencial de etiquetado de ADN de células dérmicas y epidérmicas en los injertos epidérmicos y de espesor total, sugieren una asociación con el comportamiento celular anormal, e incluso pleomórfico epidérmico, debido a la interferencia de los factores dérmicos/epidérmicos interactuantes.

Lower growth factor expression in follicular fluid undergone in-vitro fertilization / 대한생식의학회지

OBJECTIVE: This study was performed to identify whether growth and differentiation factor-9 (GDF-9) and transforming growth factor-beta1 (TGF-beta1) expressions would be lower in the follicular fluid (FF) of those over age 35 who underwent IVF than under age 35. METHODS: A total of 24 IVF cycles (20 patients) were included in this study. All of patients were stimulated for IVF by the GnRH short protocol and divided into two groups for analysis, according to their age: or =35 group (10 cycles, 9 patients). The expression levels of GDF-9 and TGF-beta1 were determined by western blotting and quantitative enzyme-linked immunosorbent assay. RESULTS: The numbers of retrieved oocytes and metaphase II oocytes were significantly lower in the > or =35 group. Lower expression of GDF-9 and TGF-beta1 by western blotting in the > or =35 group were observed as well. The mean GDF-9 and TGF-beta1 levels by enzyme-linked immunosorbent assay were lower in the > or =35 group. The values were 6,850.5+/-928.4 ng/L vs. 3,333.3+/-1,089.2 ng/L of GDF-9 (p<0.05) and 3,844.1+/-571.1 ng/L vs. 2,187.7+/-754.0 ng/L of TGF-beta1 (p<0.05). A negative correlation between GDF-9 and age was observed (r=-0.546, p=0.006). CONCLUSION: GDF-9 and TGF-beta1 production from stimulated ovaries during IVF appears to decrease with age.

Effects of nitric oxide on the proliferation and differentiation of human periodontal ligament cells / 대한치과교정학회지

OBJECTIVE: This study evaluated the effects of nitric oxide (NO) on the proliferation and differentiation of human periodontal ligament cells involved in orthodontic tooth movement. METHODS: A range of concentrations of sodium nitroprusside (SNP), a NO donor, were administered to samples of human periodontal ligament cells, followed by measurement of cell viability, alkaline phosphatase (ALP) activity, and expression of osteonectin and bone sialoprotein. RESULTS: Cell viability, ALP activity and the expression of osteonectin and bone sialoprotein were increased in human periodontal ligament cells treated with SNP concentrations of 1.0 mM. CONCLUSION: NO has a biphasic effect on proliferation and differentiation in human periodontal ligament cells, with a stimulatory effect at low concentrations and an inhibitory effect at high concentrations.

Effect of IL-6、IL-1?、TNF? on the Growth of Human Fetal Cerebral Neurons in vitro / 中国免疫学杂志

Objective:To study the effects of Il 6?IL 1??TNF? on the growth and development of cultured human fetal cerebral neurons.Methods:Established the model of primary culture of human fetal cerbral neurons in SFFD,by morphological observation and MTT assay.Results:IL 6 and IL 1? both promote the survival of neurons and IL 6 also promote the outgrowth of neuritis and differentiation of neurons in culture.TNF ? continuously inhibit the survival of neurons.Conclusion:IL 6?IL 1??TNF? can effect the development of cultured neurons.IL 6 and IL 1? both may effect as neurotrophic factor in the survival?growth and development by indirect mechanism.TNF? directly show neurotoxin effect in culture.

Effects of Polylysine Coating on GH3 Rat Pituitary Tumor Cells in Culture / 대한해부학회지

GH3 cells are derived from rat pituitary tumor cells that secrete prolactin and growth hormone, and important in studying prolactin-secreting pitutary tumors. This study was performed to examine the effects of polylysine on growth and differentiation of GH3 cells by means of (a) cell attachment assay (b) cell count and bromodeoxyuridine labeling and (c) immunohistochemistry for prolactin in the absence or presence of epidermal growth factor (EGF). Cell shape, attachment to the culture surface and growth of GH3 cells were not affected by polylysine coating. The percentages of prolactin-immunoreactive cells were higher in the cells cultured on the polylysine-coated surface compared to those on the plastic surface. Cell number and BrdU incorporation were lower in the EGF-treated cells on both culture surfaces. The results provided basic information on the effects of polylysine coating on GH3 cells in culture and suggested that polylysine coating was useful for the study on GH3 cells because it enhanced cell differentiation as well as it provided stronger attachment than plastic surfaces.