Effects of hypoxia-inducible factors on tumor mesenchymal cells: a review / 预防医学

@#Hypoxia is the most common tumor microenvironment caused by rapid proliferation of tumor cells, and hypoxia-inducible factor (HIF) is the main transcription factor for tumor cells to adapt to hypoxia. Current research has found that HIF can interact with a variety of mesenchymal cells such as fibroblasts, endothelial cells and immune cells in the tumor microenvironment, leading to the transcription and expression of target genes in response to hypoxia, which ultimately promotes tumor angiogenesis, and induces physiological changes such as migration, invasion, and immune escape of tumor cells. However, the signaling pathways involved in the HIF regulatory mechanism are complex, and the mechanism of HIF in the tumor microenvironment need to be further investigated, also most HIF inhibitors are still in the preclinical research stage. This paper reviews the research progress on the effects of HIF on tumor mesenchymal stromal cells to provide a theoretical basis for the diagnosis, prevention and treatment of tumors targeting HIF.

Therapeutic potential of human umbilical cord–derived mesenchymal stem cells transplantation in rats with optic nerve injury

Purpose: There are no effective treatments currently available for optic nerve transection injuries. Stem cell therapy represents a feasible future treatment option. This study investigated the therapeutic potential of human umbilical cord–derived mesenchymal stem cell (hUC?MSC) transplantation in rats with optic nerve injury. Methods: Sprague–Dawley (SD) rats were divided into three groups: a no?treatment control group (n = 6), balanced salt solution (BSS) treatment group (n = 6), and hUC?MSCs treatment group (n = 6). Visual functions were assessed by flash visual evoked potential (fVEP) at baseline, Week 3, and Week 6 after optic nerve crush injury. Right eyes were enucleated after 6 weeks for histology. Results: The fVEP showed shortened latency delay and increased amplitude in the hUC?MSCs treated group compared with control and BSS groups. Higher cellular density was detected in the hUC?MSC treated group compared with the BSS and control groups. Co?localized expression of STEM 121 and anti?S100B antibody was observed in areas of higher nuclear density, both in the central and peripheral regions. Conclusion: Peribulbar transplantation of hUC?MSCs demonstrated cellular integration that can potentially preserve the optic nerve function with a significant shorter latency delay in fVEP and higher nuclear density on histology, and immunohistochemical studies observed cell migration particularly to the peripheral regions of the optic nerve.

Review of factors in pterygium pathology / 国际眼科杂志(Guoji Yanke Zazhi)

Pterygium is an ocular surface disease formed by many factors and associate with a series of changes caused by ultraviolet irradiation and radiation, its pathogenesis is still uncertain. Elevated vascular endothelial growth factor(VEGF), inflammatory infiltrates, angiogenesis, oxidative stress, epithelial-mesenchymal cell transition(EMT), and tumor suppressor gene inactivation are currently recognized causes of pterygium. The mechanism of this factor in pterygium deveopment is still not completely understood. This review aimed to investigate the role of these factors in pterygium formation and provide targeted therapy and effective preventive measures for clinical diagnosis and treatment.

Effects of bone marrow mesenchymal cells from immune thrombocytopenia patients on the biological behaviors of megakaryocytes / 上海交通大学学报（医学版）

Objective • To investigate changes of immune thrombocytopenia (ITP) patients-derived bone marrow mesenchymal cells (BMCs) in cells survival, cytokines expression as well as the effects of BMCs on the biological behaviors of megakaryocytes. Methods • BMCs were collected from 7 ITP patients and 5 normal controls (NC), and cultivated by the whole marrow adherent method. Surface markers and basal apoptosis rate of BMCs were analyzed by flow cytometry (FCM). Proliferation of BMCs was assessed by CCK-8 method. Phorbol 12-myristate 13-acetate (PMA) was used to stimulate differentiation of HEL cells. The induced HEL cells (inHEL) were divided into 3 groups: inHEL cultured alone (group a), inHEL co-cultured with BMCs derived from ITP patients (group b), inHEL co-cultured with BMCs derived from NC (group c). After 72 h incubation, the expression of cell surface proteins (CD41a, CD42b) and cell apoptosis rate were analyzed by FCM. The mRNA and proteins expression levels of cytokines IL6, IL11, TPO, SCF were detected by real-time fluorescent quantitative PCR (RT-qPCR) and enzyme linked immunosorbent assay (ELISA), respectively. Results • Compared with NC, BMCs from ITP patients grew progressively slowly (Day 4, P=0.039; Day 6, 10, P=0.009; Day 8, P=0.007), cell basal apoptosis rates were increased [AV+PI- (early apoptosis rate), P=0.036; AV+PI+ (late apoptosis rate), P=0.003; AV+PI-/+ (total apoptosis rate), P=0.004]. Compared with group a, the expression of CD41a in group c was much higher (P=0.000). The expression of CD41a in group b was higher than that in group a (P=0.015), but still much less than that in group c (P=0.000). Compared with group a, the early and total apoptosis rate in group b, c and the late apoptosis rate in group c were decreased obviously (all P=0.000), whereas there was no obvious change of the late apoptosis rate in group b. However, compared with group c, the late and total apoptosis rate in group b were significantly increased (both P=0.000). The expression levels of IL6, SCF mRNA and IL6 protein were significantly decreased in ITP BMCs (all P=0.000), but there was no obvious difference in the expression levels of IL11 and TPO between ITP BMCs and NC BMCs. Conclusion • BMCs from ITP patients show some defects in supporting megakaryocytic differentiation and survival under co-culture conditions, which mechanisms are related to the reduction of IL6 and SCF expression.

Effects of bone marrow mesenchymal cells from immune thrombocytopenia patients on the biological behaviors of megakaryocytes / 上海交通大学学报（医学版）

Objective·To investigate changes of immune thrombocytopenia (ITP) patients-derived bone marrow mesenchymal cells (BMCs) in cells survival, cytokines expression as well as the effects of BMCs on the biological behaviors of megakaryocytes. Methods?·?BMCs were collected from 7 ITP patients and 5 normal controls (NC), and cultivated by the whole marrow adherent method. Surface markers and basal apoptosis rate of BMCs were analyzed by flow cytometry (FCM). Proliferation of BMCs was assessed by CCK-8 method. Phorbol 12-myristate 13-acetate (PMA) was used to stimulate differentiation of HEL cells. The induced HEL cells (inHEL) were divided into 3 groups: inHEL cultured alone (group a), inHEL co-cultured with BMCs derived from ITP patients (group b), inHEL co-cultured with BMCs derived from NC (group c). After 72 h incubation, the expression of cell surface proteins (CD41a, CD42b) and cell apoptosis rate were analyzed by FCM. The mRNA and proteins expression levels of cytokines IL6, IL11, TPO, SCF were detected by real-time fluorescent quantitative PCR (RT-qPCR) and enzyme linked immunosorbent assay (ELISA), respectively. Results?·?Compared with NC, BMCs from ITP patients grew progressively slowly (Day 4, P=0.039; Day 6, 10, P=0.009; Day 8, P=0.007), cell basal apoptosis rates were increased [AV+PI- (early apoptosis rate), P=0.036; AV+PI+(late apoptosis rate), P=0.003; AV+PI-/+(total apoptosis rate), P=0.004]. Compared with group a, the expression of CD41a in group c was much higher (P=0.000). The expression of CD41a in group b was higher than that in group a (P=0.015), but still much less than that in group c (P=0.000). Compared with group a, the early and total apoptosis rate in group b, c and the late apoptosis rate in group c were decreased obviously (all P=0.000), whereas there was no obvious change of the late apoptosis rate in group b. However, compared with group c, the late and total apoptosis rate in group b were significantly increased (both P=0.000). The expression levels of IL6, SCF mRNA and IL6 protein were significantly decreased in ITP BMCs (all P=0.000), but there was no obvious difference in the expression levels of IL11 and TPO between ITP BMCs and NC BMCs. Conclusion?·?BMCs from ITP patients show some defects in supporting megakaryocytic differentiation and survival under co-culture conditions, which mechanisms are related to the reduction of IL6 and SCF expression.

Células madre: buscando marcadores de superficie celular que predispongan compromiso de diferenciación cardiaca / Stem cells: searching predisposition to cardiac commitment by surface markers expression

Resumen Actualmente las enfermedades cardiovasculares se han convertido en un serio problema para los sistemas de salud de todo el mundo, ya que son la principal causa de muerte y representan una enorme carga económica. Este problema ha sido abordado con diferentes estrategias, entre ellas con la ayuda de terapia celular, aunque sin resultados contundentes. Durante más de 20 años, se ha utilizado una gran variedad de células madre en diferentes modelos de infarto del miocardio. El uso de células madre cardiacas (CSC) parece ser la mejor opción, pero la inaccesibilidad y la escasez de estas células hacen que su uso sea muy limitado. Además, existe un riesgo elevado pues tienen que obtenerse directamente del corazón del paciente. A diferencia de las CSC, las células madre adultas derivadas de médula ósea o tejido adiposo, entre otras, representan una opción atractiva debido a su fácil accesibilidad y abundancia, pero sobre todo a la probable existencia de progenitores cardiacos entre sus diferentes subpoblaciones. En esta revisión hacemos un análisis de los marcadores de superficie presentes en CSC en comparación con otras células madre adultas, y sugerimos la preexistencia de células que comparten marcadores de superficie específicos con CSC, la presencia de un inmunofenotipo predecible, aunque en proporciones bajas, pero con un potencial de diferenciación cardiaca similar a las CSC, lo cual podría aumentar su valor terapéutico. Este estudio revela las nuevas perspectivas con respecto a la presencia de dichos marcadores, los cuales comprometerían algunas de estas subpoblaciones a diferenciarse a tejido cardiaco.

Abstract It is well-known that cardiovascular diseases are the leading cause of death world- wide, and represent an important economic burden to health systems. In an attempt to solve this problem, stem cell therapy has emerged as a therapeutic option. Within the last 20 years, a great variety of stem cells have been used in different myocardial infarction models. Up until now, the use of cardiac stem cells (CSCs) has seemed to be the best option, but the inaccessibility and scarcity of these cells make their use unreliable. Additionally, there is a high risk as they have to be obtained directly from the heart of the patient. Unlike CSCs, adult stem cells originating from bone marrow or adipose tissue, among others, appear to be an attractive option due to their easier accessibility and abundance, but particularly due to the probable existence of cardiac progenitors among their different sub-populations. In this review an analysis is made of the surface markers present in CSCs compared with other adult stem cells. This suggested the pre-existence of cells sharing specific surface markers with CSCs, a predictable immunophenotype present in some cells, although in low proportions, and with a potential of cardiac differentiation that could be similar to CSCs, thus increasing their therapeutic value. This study highlights new perspectives regarding MSCs that would enable some of these sub-populations to be differentiated at cardiac tissue level.

Effect of bone mesenchymal stem cell transplantation on accelerating the vascularization surrounding transplant pancreatic islet / 器官移植

Objective To investigate whether pancreatic islet transplantation in combination with bone mesenchymal stem cells (MSC) transplantation can promote the vascularization surrounding the transplant pancreatic islet.Methods The non-obese diabetic (NOD) mice were utilized as the recipients and randomly divided into pancreatic islet transplantation combined with MSC transplantation group (co-transplantation group,n=6),pancreatic islet transplantation group(n=6),MSC transplantation group(n=6) and sham transplantation group (n=3).The variation in blood glucose level and survival rate post-transplantation of NOD mice in each group was observed.The proliferation and apoptosis of the transplant pancreatic islet in the pancreatic islet transplantation group and co-transplantation group at 1,2 and 4 weeks after pancreatic islet transplantation were analyzed by 5-ethynyl-2'-deoxyufidine (EdU) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).The vascular density surrounding the transplanted pancreatic islet in the pancreatic islet transplantation group and co-transplantation group at postoperative 2,4 and 8 weeks were observed under light microscope and quantitatively analyzed by histochemical and immunohistochemical staining.Results Both MSC combined with pancreatic islet transplantation and pancreatic islet transplantation significantly improved the blood glucose level and enhanced the survival rate of NOD mouse models after transplantation.In addition,it could accelerate the regeneration of pancreatic islet cells and decrease cell apoptosis.MSC combined with pancreatic islet transplantation significantly enhanced the vascular density surrounding the transplant pancreatic islet compared with pancreatic islet transplantation alone.Conclusions MSC transplantation can accelerate the vascularization surrounding the transplant pancreatic islet,increase the blood supply and protect the function and activity of the transplant pancreatic islet.

Combination of musk and olibanum promotes proliferation of prostate stem cells in mice: An experimental study / 中华男科学杂志

Objective@#To study the effects of muskolibanum combination on the proliferation and differentiation of prostate stem cells.@*METHODS@#We cultured prostate epithelial cells and urogenital sinus mesenchymal (UGSM) cells from 7－10 d old C57BL/6 mice and 16－18 d old pregnant C57BL/6 mice, transplanted the mixed suspension of the two types of cells under the kidney envelope of SCIDCB.17 male mice, and harvested the transplants 30 days later. We randomly divided the SCIDCB.17 mice into four groups to be treated intragastrically with musk (n = 8), olibanum (n = 8), musk+olibanum (n = 7), and normal saline (blank control, n = 8)) respectively, all for 14 days. Then we collected the kidney tissue for observation of the morphology of the glandular tubes and differentiation of different subsets of stem cells by HE staining and determination of the expressions and distribution of P63, CD133, CD117 and Sca1 by immunohistochemistry and Western blot.@*RESULTS@#A system was successfully established for the isolation and mixed culture of Sca1 Lin+ CD49f+ (LSC) cells of prostate stem cells and UGSM cells of the mouse embryonic prostate. Immunohistochemistry showed positive expressions of P63, CD133, Sca1, and CD117 in the prostatic acinar epithelia and proved the presence of prostatic acinar epithelial structure in the transplants. Compared with the blank control group, the expressions of CD133, Sca1 and CD117 were significantly increased in the musk, olibanum, and musk+olibanum groups (P< 0.05), higher in the musk+olibanum than in the musk or olibanum group (P< 0.05), and their protein expressions were even more elevated in the musk+olibanum group (P< 0.01), with statistically significant difference from the olibanum group (P< 0.05).@*CONCLUSIONS@#The combination of musk and olibanum can improve the proliferation and differentiation of prostate stem cells.

Immediate and delayed intracavernous injection of bone marrow mesenchymal stem cells to improve erectile function in rats with cavernous nerve injury / 中华男科学杂志

Objective@#To explore the effects of immediate and delayed intracavernous injection of bone marrow mesenchymal stem cells (BM-MSCs) on neurogenic erectile dysfunction (NED) induced by bilateral cavernous nerve injury in Sprague-Dawley (SD) rats.@*METHODS@#BM-MSCs isolated from male SD rats were cultured and identified. Twenty-eight 8-week-old male SD rats were randomly divided into four groups, sham operation, NED model control, BM-MSCs immediate, and BM-MSCs delayed, and NED models were established in the latter three groups by crushing the bilateral cavernous nerves. The rats in the sham operation and model control groups were injected intracavernously with placebo while those in the latter two with BM-MSCs immediately or 2 weeks after modeling. At 12 weeks after operation, the penile function of the rats was assessed according to the penile intracavernous pressure (ICP), mean arterial pressure (MAP), and ICP/MAP ratio obtained from different groups of rats. Then, all the animals were sacrificed and the penile cavernosal tissue collected for histological analysis.@*RESULTS@#At 12 weeks after modeling, both ICP and ICP/MAP were significantly increased in the BM-MSCs immediate and delayed groups as compared with those in the model control (P <0.05), and so were the ratio of smooth muscle to collagen (P <0.05) and the smooth muscle content in the corpus cavernosum (P <0.05), and the number of neurofilament (NF)-positive nerve fibers (P <0.05) and the expression of neuronal nitric oxide synthase (nNOS) in the dorsal nerves of the midshaft penis (P <0.05).@*CONCLUSIONS@#Intracavernous injection of BM-MSCs can improve erectile function in rats with bilateral cavernous nerve injury by elevating the smooth muscle-collagen ratio and smooth muscle content in the corpus cavernosum and thus preventing its fibrosis as well as by increasing the number of NF-positive nerve fibers and expression of nNOS in the penile dorsal nerves.

Effects of bone marrow mesenchymal stem cells transplantation on the neurotrophic factor expression and cell apoptosis in the rats with spina bifida aperta / 国际儿科学杂志

Objective To observe the effects of bone marrow mesenchymal stem cells (BMSC) transplantation on the expression of nerve growth factor(NGF) and brain derived neurotrophic factor(BDNF) in the rat spinal cord with spina bifida,and to investigate the change in cell apoptosis after BMSC transplantation.Methods Spina bifida aperta was induced with a single intragastric injection of all-trans retinoic acid,then the BMSC was microinjected into spina cord of rat embryos on embryo 16 d(E16),BDNF and NGF were tested by immunofluorescence staining,and TUNEL assay were used for investigating cell apoptosis.Results Transplantation of BMSC enhanced the expression of NGF and BDNF,and reduced cell apoptosis in the defective site of spinal cord.Conclusion The transplantation of BMSC may improve the microenvironment of spinal cord and repair the neurological defects by enhancing the expression of neurotrophic factor and reducing the cell apoptosis.

Vit D promotes proliferation and differentiation of mesenchymal stem cells through regulates extracts of plastrum testudinis / 局解手术学杂志

Objective To explore the effect and the mechanism of vitamin D(Vit D) promotes proliferation and differentiation of mesenchymal stem cells (MSCs) through regulates extracts of plastrum testudinis (PTE).Methods Established the PGL3-Id1 promoter and transfered rat MSCs.PTE combined with 10-6,10-7,10-8mol/L Vit D respectively were acted on the transfected MSCs for 36 hours.The level of Id1 promoter were detected by luciferase activity measurement.1,3,30,100 pg/mL PTE combined with Vit D of 10-7 mol/L were acted on MSCs for 36 hours,3 days and 7 days,and the VDR expression were detected by RT-PCR test.Results PTE promoted the expression of Id1 in MSCs,the expression of Id1 was inhibited when PTE combined with Vit D (P ＜ 0.01),and it was significantly different among different dosis of Vit D(P ＜0.01).The expression of VDR was inhibited in different degree when PTE combined with Vit D for 36 hours,3 days and 7 days.PTE combed with large dose of Vit D for 36 hours had significant effect of inhibition,and the difference was statistically significant (P ＜ 0.05).The inhibiting effect was more obvious when PTE combined with large dose of Vit D for 3 days and 7 days.When different doses of PTE combined with Vit D for a same duration,the difference of VDR expression was statistically significant (P ＜ 0.05).Meanwhile,when same doses of PTE combined with Vit D for different durations,the difference of VDR expression at 7 days and 36 hours was statistically significant (P ＜ 0.05).Conclusion The proliferation of MSCs which promoted by PTE was inhibited by Vit D,and the nuclear receptor VDR may be one of the targets of drug action for PTE regulating proliferation and differentiation of MSCs.

Autologous Bone Marrow Mesenchymal Cell Induced Chondrogenesis for the Treatment of Osteoarthritis of Knee

Healthy and high quality of life has become the main issue with increasing human life span. Many biological treatments for osteoarthritis of the knee have been tried with limited success. We compared data from 7 patients who underwent total knee arthroplasty and 46 patients who underwent autologous bone-marrow mesenchymal cell induced chondrogenesis (MCIC) for osteoarthritis of grade IV of the Kellgren-Lawrence classification and grade IV of modified Outerbridge classification from 50 to 65 years of age. Clinical evaluation of the 2 groups showed significant improvement in the mean telephone Knee Society Scoring system (tKSS)-A (pain) and tKSS-B (function) scores throughout the postoperative follow-up period. There was no difference in the patients' satisfaction between the 2 groups. MCIC is a treatment option at least for delaying disease progression of osteoarthritis of the knee.

Prostaglandin E2 induced myofibrolast phenotype inversion and interleukin 8 expression in prostate stromal cells / 国际检验医学杂志

Objective To investigate the effects of prostaglandin E2(PGE2) in prostate mesenchymal cell activation .Methods To culture human prostate stromal cells WPMY‐1 in vitro ,implement immunofluorescence staining after treating with DMSO and 10-9 mol/L PGE2 and detect the change of the myofibroblast phenotype .The expression of cellular inflammatory factor interleukin 8(IL‐8) was detected by real‐time quantitative PCR and the concentration of IL‐8 was detected .Results PGE2 significantly in‐creased the cellular proportion of colocalization with alpha SMA and Vimentin in WPMY‐1 cells .PGE2 promoted the expression of IL‐8 in WPMY‐1 cells .Conclusion PGE2 can increase myofibroblast phenotype in human prostate mesenchymal cells WPMY‐1 in vitro culture ,promotes the IL‐8 expression ,which has an important role in the occurrence and development of prostate cancer .

A Case of Pulmonary Inflammatory Pseudotumor / 소아알레르기및호흡기학회지

Inflammatory pseudotumor of the lung is considered to be a rare, benign, neoplastic lesion, consisting mainly of spindle mesenchymal cells, sometimes in such a way that its histological appearance mimics that of a spindle cell sarcoma, fibrous histiocytoma or fibrosarcoma. A case of inflammatory pseudotumor of the lung occurring in a 13-year-old boy is reported with pathologic findings, including its ultrastructure. The patient had had no symptoms and accidentally discovered his condition after a chest X-ray examination at a regular school physical check up. The mass was located in the suprahilar area of the left lung. Exploratory thoracotomy revealed a large mass that was removed, together with the left upper lobe of the lung. Microscopically, the mass was composed of numerous interstitial inflammatory cells, mainly lymphoplasma cells. Ultrastructurally, the spindle-shaped mesenchymal cells were arranged haphazadly and the normal pulmonary structure was nearly totally destroyed. Emphasis is given to complete resection of the tumor for both diagnostic and therapeutic purposes.

In vitro neuronal and osteogenic differentiation of mesenchymal stem cells from human umbilical cord blood

Mesenchymal stem cells (MSCs) have the capabilities for self-renewal and differentiation into cells with the phenotypes of bone, cartilage, neurons and fat cells. These features of MSCs have attracted the attention of investigators for using MSCs for cell-based therapies to treat several human diseases. Because bone marrowderived cells, which are a main source of MSCs, are not always acceptable due to a significant drop in their cell number and proliferative/differentiation capacity with age, human umbilical cord blood (UCB) cells are good substitutes for BMCs due to the immaturity of newborn cells. Although the isolation of hematopoietic stem cells from UCB has been well established, the isolation and characterization of MSCs from UCB still need to be established and evaluated. In this study, we isolated and characterized MSCs. UCB-derived mononuclear cells, which gave rise to adherent cells, exhibited either an osteoclast or a mesenchymal-like phenotype. The attached cells with mesenchymal phenotypes displayed fibroblast-like morphologies, and they expressed mesenchymal-related antigens (SH2 and vimentin) and periodic acid Schiff activity. Also, UCB-derived MSCs were able to transdifferentiate into bone and 2 types of neuronal cells, in vitro. Therefore, it is suggested that the MSCs from UCB might be a good alternative to bone marrow cells for transplantation or cell therapy.

Differentiation of Human Amniotic Mesenchymal Cells into Cardiomyocyte-like Cells in Vitro / 中国生物工程杂志

To explore the plasticity of human amniotic mesenchymal cells(hAMCs) into cardiomyocyte-like cells,hAMCs were isolated from human amnion with collagenase digestion.Phenotype of the isolated cells was analyzed by flow cytometry(FCM).hAMCs were treated with 5-azacytidine and basic fibroblast growth factor(bFGF) to investigate their ability of differentiation into cardiomyocytes.The induced differentiated cells were evaluated by immunofluorescence for desmin and ?-actin expression and by RT-PCR for Nkx2.5,GATA-4 and alpha-myosin heavy chain(?-MHC) mRNA expression.The results showed that,after primary culture,hAMCs could reach a confluence of 80% with swirl like growth at 6 days.The cells proliferated rapidly after passages with a 100% confluence at 3~4 d.hAMCs were positive expression of CD44 and vimentin,but negative for CK19.After induced differentiation at 8~10d,the differentiated cells have close-up arranged with long spindle-shape.At 2 weeks and 4 weeks,induced cells expressed ?-actinin and cardiac-specific transcription factor Nkx2.5.Expressions of GATA-4 and desmin can be detected but ?-MHC can not in the hAMCs both before and after the induction.In conclusion,hAMCs have the ability of differentiation into cardiomyocyte-like cells,which means that hAMCs can be regarded as candidate cells for cellular cardiomyoplasty(CCM).

Development of a bioactive cellulose membrane from sea squirt skin for bone regeneration - a preliminary research

OBJECTIVES: To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. MATARIALS AND METHODS: After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with HandE and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. RESULTS: The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. CONCLUSION: We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.

The signal transduction and drug development of epithelial-mesenchymal transition / 中国药理学通报

Epithelial-mesenchymal-transition(EMT) plays a key role in the formation of embryo.At present,it is believed that EMT also contribute to the metastasis of primary tumors.A lot of signaling pathways participate in EMT,such as Wnt/?-catenin,Notch,Hedgehog,TGF? and growth factor receptors.The drugs that affect these pathways may play an important part in oncotherapy.

The Effect Of Hyaluronic Acid On Mouse Calvaria Pre-Osteoblasts Osteogenesis In Vitro

Hyaluronic acid (HA) is an almost essential component of extracellular matrices. Early in embryogenesis mesenchymal cells migrate, proliferate and differentiate, in part, because of the influence of HA. Since the features of embryogenesis are revisited during wound repair, including bone fracture repair, this study was initiated to evaluate whether HA has an effect on calcification and bone formation in an in vitro system of osteogenesis. Mouse calvaria Pre-osteoblast (MC3T3-E1) cells were cultured in alpha-MEM medium with microorganism-derivative hyaluronic acid that was produced by Strep. zooepidemicus which of molecular weight was 3 million units. The dosages were categorized in each 0.5, 1.0 and 2.0 mg/ml concentration experimental groups. After 2 and 4 days cultures in expeirmental and control groups, the tendency of cell proliferation, MTT assay, protein synthesis ability, collagen synthesis and alkaline phosphatase activity were analysed and bone nodule formation capacity were measured with Alizarin Red S stain after 29 days cultures. The cell proliferation was increased in time, especially the group of 0.5 and 1.0 mg/ml concentration of HA were showed prominent cell proliferation. After 2 and 4 days culture, experimental groups in general were greater cell activity in MTT assay. The protein synthesis was increased in all experimental groups compared to control group, especially most prominent in 1.0 mg/ml concentration group. The collagen synthesis capacity were increased in HA experimental groups, especially prominent in 1.0 mg/ml group and the activity of alkaline phosphatase were increased, especially also prominent in 1.0 mg/ml group, compared to control group. Above these, the activity of mouse carvarial pre-osteoblast cells was showed greater bone osteogenesis activity in all applied HA experimental group, especially group of 1.0 mg/ml concentration of HA.

Differentiation of mesenchymal cells derived from mouse fetal liver into neuronal cells in vitro / 第二军医大学学报

Objective:To investigate potency of mouse fetal liver mesenchymal cells differentiating into neural cells in vitro .Methods: Sterile cells from 14.5 day old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20%FCS and adherent cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by ? mercaptoethanol (? ME) and all trans retinoic acid (ATRA). The characteristics of treated cells were assayed by immunocytochemistry staining and semi quantitative RT PCR method on 5 h and 5 d after induction. Results: After induction by ? ME and ATRA, approximately 80% of the cells exhibited typical neuronal morphology;most cells expressed neuronal phenotype such as neuron specific nuclear protein(NeuN), neuron specific tubulin Ⅲ(TuJ 1), neurofilament M(NF M),and neuron specific enolase(NSE).However, microtubule associated protein tau(Tau 1)and microtubule associated protein 2 (MAP 2) as indicators of mature neuronal phenotype were undetectable. Semi quantitative RT PCR showed increased mRNA expression of brain factor 1(BF 1), brain 3(Brn 3), and neurofilament L(NF L) in treated cells versus untreated cells ( P