Size-dependent biological effects on vascular endothelial cells induced by different particulate matters

Summary： The contribution of particles to cardiovascular mortality and morbidity has been enlightened by epidemiologic and experimental studies. However, adverse biological effects of the particles with different sizes on cardiovascular cells have not been well recognized. In this study, sub-cultured human umbilical vein endothelial cells （HUVECs） were exposed to increasing concentrations of pure quartz particles （DQ） of three sizes （DQPM1, 〈1 μm; DQPM3-5, 3-5 μm; DQPM5, 5 μm） and carbon black particles of two sizes （CB0.1, 〈0.1 μm; CB 1, 〈 1 μm） for 24 h. Cytotoxicity was estimated by measuring the activity of lactate dehydrogenase （LDH） and cell viability. Nitric oxide （NO） generation and cyto- kines （TNF-α and IL-1β） releases were analyzed by using NO assay and enzyme-linked immunoabsorbent assay （ELISA）, respectively. It was found that both particles induced adverse biological effects on HUVECs in a dose-dependent manner. The size of particle directly influenced the biological activity. For quartz, the smaller particles induced stronger cytotoxicity and higher levels of cytokine responses than those particles of big size. For carbon black particles, CB0.1 was more capable of inducing adverse responses on HUVECs than CB 1 only at lower particle concentrations, in contrast to those at higher concentrations. Meanwhile, our data also revealed that quartz particles performed stronger cell damage and produced higher levels of TNF-α than carbon black particles, even if particles size was similar. In conclusion, particle size as well as particle composition should be both considered in assessing vascular endothelial cells injury and inflammation responses induced by particles.     

Mouse A6-positive Hepatic Oval Cells Derived from Embryonic Stem Cells简

Summary： Oval cells have a potential to differentiate into a variety of cell lineages including hepatocytes and biliary epithelia. Several models have been established to activate the oval cells by incorporating a variety of toxins and carcinogens, alone or combined with surgical treatment. Those models are obviously not suitable for the study on human hepatic oval cells. It is necessary to establish a new and efficient model to study the human hepatic oval cells. In this study, the hepatocyte growth factor（HGF） and epidermal growth factor（EGF） were used to induce differentiation of mouse embryonic stem（ES） cells into hepatic oval cells. We first confirmed that hepatic oval cells derived from ES cells, which are bipotential, do exist during the course of mouse ES cells＇ differentiation into hepatic parenchymal cells. RT-PCR and transmission electron microscopy were applied in this study. The ratio of Sca-1＋/CD34＋ cells sorted by FACS in the induction group was increased from day 4 and reached the maximum on the day 8, whereas that in the control group remained at a low level. The differentiation ratio of Sca-1＋/CD34＋ cells in the induction group was significantly higher than that in the control group. About 92.48% of the sorted Sca-1＋/CD34＋ cells on the day 8 were A6 positive. Highly purified A6＋/Sca-1＋/CD34＋ hepatic oval cells derived from ES cells could be obtained by FACS. The differentiation ratio of hepatic oval cells in the induction group（up to 4.46%） was significantly higher than that in the control group. The number of hepatic oval cells could be increased significantly by HGF and EGF. The study also examined the ultrastructures of ES-derived hepatic oval cells＇ membrane surface by atomic force microscopy. The ES-derived hepatic oval cells cultured and sorted by our protocols may be available for the future clinical application.     

Human adipose-derived mesenchymal stem cells: a better cell source for nervous system regeneration

Background In order to suggest an ideal source of adult stem cells for the treatment of nervous system diseases,MSCs from human adipose tissue and bone marrow were isolated and studied to explore the differences with regard to cell morphology,surface markers,neuronal differentiation capacity,especially the synapse structure formation and the secretion of neurotrophic factors.Methods The neuronal differentiation capacity of human mesenchymal stem cells from adipose tissue （hADSCs） and bone marrow （hBMSCs） was determined based on nissl body and synapse structure formation,and neural factor secretion function.hADSCs and hBMSCs were isolated and differentiated into neuron-like cells with rat brain-conditioned medium,a potentially rich source of neuronal differentiation promoting signals.Specific neuronal proteins and neural factors were detected by immunohistochemistry and enzyme-linked immunosorbent assay analysis,respectively.Results Flow cytometric analysis showed that both cell types had similar phenotypes.Cell growth curves showed that hADSCs proliferated more quickly than hBMSCs.Both kinds of cells were capable of osteogenic and adipogenic differentiation.The morphology of hADSCs and hBMSCs changed during neuronal differentiation and displayed neuronlike cell appearance after 14 days＇ differentiation.Both hADSCs and hBMSCs were able to differentiate into neuron-like cells based on their production of neuron specific proteins including β-tubulin-Ⅲ,neuron-specific enolase （NSE）,nissl bodies,and their ability to secrete brain derived neurotrophic factor （BDNF） and nerve growth factor （NGF）.Assessment of synaptop hysin and growth-associated protein-43 （GAP-43） suggested synapse structure formation in differentiated hADSCs and hBMSCs.Conclusions Our results demonstrate that hADSCs have neuronal differentiation potential similar to hBMSC,but with a higher proliferation capacity than hBMSC.Adipose tissue is abundant,easily available and would be a potential ideal source of adult stem cells for neural-related clinical research and application.     

New insights into the effects of Mycobacterium bovis Bacillus Calmette-Guérin on asthma

Allergic asthma has been defined as a disease of immunodysregulation, where the pathology is a direct consequence of excessive T-helper type 2 immune responses （Th2） to allergens in the lungs. Disease is associated with increased secretion of type-2 cytokines such as interleukin （IL）-4, IL-5 and IL-13 which, among other things,     

Application of tissue engineering in the treatment of stress urinary incontinence

Stress urinary incontinence is one of the most common diseases in urology. The main treatments for stress urinary incontinence are pharmacotherapy, physicobehavioral therapy and surgery.However, the results of present methods are not satisfactory. Tissue engineering is a newly emerging technology that may provide a novel method for the treatment of stress urinary incontinence.     

Effect of Intracoronary Infusion of Bone Marrow Mononuclear Cells or Peripheral Endothelial Progenitor Cells on Myocardial Ischemia-reperfusion Injury in Mini-swine

Objective To simulate and assess the clinical effect of intracoronary infusion of bone marrow mononuclear cells or peripheral endothelial progenitor cells on myocardial reperfusion injury in mini-swine model. Methods Twenty-three mini-swine with myocardial reperfusion injury were used as designed in the study protocol. About （3.54±0.90）×10^7 bone marrow mononuclear cells （MNC group, n=9） or （1.16± 1.07）× 10^7 endothelial progenitor cells （EPC group, n=7） was infused into the affected coronary segment of the swine. The other mini-swine were infused with phosphate buffered saline as control （n=7）. Echocardio- graphy and hemodynamic studies were performed before and 4 weeks after cell infusion. Myocardium infarc- tion size was calculated. Stem cell differentiation was analyzed under a transmission electromicroscope. Results Left ventricular ejection fraction dropped by 0% in EPC group, 2% in MNC group, and 10% in the control group 4 weeks after cell infusion, respectively （P〈0.05）. The systolic parameters increased in MNC and EPC groups but decreased in the control group. However, the diastolic parameters demonstrated no significant change in the three groups （P〉0.05）. EPC decreased total infarction size more than MNC did （1.60±0.26 cm2 vs. 3.71±1.38 cm2, P〈0.05）. Undermature endothelial cells and myocytes were found under transmission electromlcroscope. Conclusions Transplantation of either MNC or EPC may be beneficial to cardiac systolic function, but might not has obvious effect on diastolic function. Intracoronary infusion of EPC might be better than MNC in controlling infarction size. Both MNC and EPC may stimulate angiogenesis, inhibit flbrogenesis, and differentiate into myocardial cells.     

Future application of hair follicle stem cells: capable in differentiation into sweat gland cells

Background Sweat glands (SGs) can not regenerate after complete destruction in the severe skin injury,so it is important to find a ideal stem cell source in order to regenerate functional SGs.Hair foll...     

Influence of sinomenine on protein profiles of peripheral blood mononuclear cells from ankylosing spondylitis patients: a pharmacoproteomics study

Background Ankylosing spondylitis （AS） is a common inflammatory rheumatic disease which lacks satisfactory treatment so far.Sinomenine （SIN） is an alkaloid and has recently been utilized in treating multiple rheumatic diseases including AS in China,but its exact mechanism remains to be explored.This study investigated the alteration of proteome in peripheral blood mononuclear cells （PBMCs） from AS patients.Methods Thirty AS patients were enrolled in this study.PBMCs from each AS patient were cultured in medium with or without SIN respectively.Then PBMCs proteins from both groups were separated by two-dimensional electrophoresis （2-DE） and analyzed by mass spectrometry （MS）.Two differentially expressed proteins were then chosen to be verified using Western blotting.Results Seven proteins,including α-synuclein （SNCA）,calmodulin （CALM）,acidic leucine-rich nuclear phosphoprotein 32 family member A （ANP32A）,chloride intracellular channel protein 1 （CLIC1）,guanine nucleotide-binding protein G（I）/G（S）/ G（T） subunit beta-1 （GNB1）,gelsolin （GSN） and histone H2B type 1-M （HISTH2BM）were over-expressed,while coronin1A （CORO1A） was under-expressed in the SIN-treated PBMCs.Further bioinformatics search indicated that the changes of SNCA,ANP32A and CLIC1 pertained to apoptosis,while changes of GSN and CORO1A were associated with both apoptosis and inhibition of immunological function.Subsequently GSN and CORO1A were selected to validate by Western blotting and the results were consistent with those of 2-DE.Conclusion There were 8 differentially expressed proteins in the SIN-treated PBMCs,which might shed some light on the mechanism of SIN in the treatment of AS.     

Cyclooxygenase-2 Blockade Inhibits Accumulation and Function of Myeloid-derived Suppressor Cells and Restores T Cell Response after Traumatic Stress

Myeloid-derived suppressor cells （MDSCs） play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 （Cox-2） contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD1 lb＋/Gr-l＋ cells, proliferation and apoptosis of CD4＋ T cells were determined. Ar- ginase activity and arginase-1 （Arg-1） protein expression of splenic CDllb＋/Gr-I＋ cells, and de- layed-type hypersensitivity （DTH） response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD1 lb＋/Gr-l＋ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD1 lb＋/Gr-l＋ ceils. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4＋ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.     

Erbin interacts with Sema4C and inhibits Sema4C-induced epithelial-mesenchymal transition in HK2 cells

Erbin, a member of Leucine-rich repeat and PDZ-containing protein family, was found to inhibit TGF-β-induced epithelial-mesenchymal transition （EMT） in our previous study. However, the mechanism of Erbin in regulating EMT is unclear. Semaphorin protein Sema4C, with PDZ binding site at C-terminal has been recognized as a positive regulator of EMT. Here, we aimed to examine the inter- action between Erbin and Sema4C. HK2 cells were treated with TGF-β1, or transfected with Erbin and （or） Sema4C. Interaction of Erbin and Sema4C was identified by immunoprecipitation. RT-PCR was used to detect the expression of Erbin and Sema4C at mRNA level after transfection. The expression levels of Erbin, Sema4C, and markers of EMT were measured by using Western blotting or ELISA. Af- ter HK2 cells were stimulated with 10 ng/mL TGF-β1 for 72 h, the protein expression levels of Erbin and Sema4C were both up-regulated, and immunoprecipitation results showed Erbin interacted with Sema4C in HK2 cells both at endogenous and exogenous levels. Furthermore, overexpression of Sema4C suppressed E-cadherin, induced vimentin and promoted fibronectin secretion, indicating Sema4C promotes the process of EMT. However, HK2 cells overexpressing Erbin were resistant to Sema4C-induced EMT. In contrast, Erbin specific siRNA promoted EMT induced by Sema4C. Taken together, these results suggest that Erbin can interact with Sema4C, and co-expression of Erbin blocks the process of Sema4C-induced EMT.     

Amplification of functional myeloid-derived suppressor cells during stem cell mobilization induced by granulocyte colony-stimulation-factor

The effects of granulocyte colony-stimulation-factor （G-CSF） on stem cell mobilization and its impact on the amplification of myeloid-derived suppressor cells （MDSCs） of donor mice were ex- amined. A mouse model of stem cell mobilization was established by consecutive subcutaneous injec- tion of 100 μg/kg G-CSF for 5 days. The blood from the donor mice was routinely examined during mobilization. Stem cells and MDSCs were analyzed by flow cytometry. The immunosuppressive mole- cules derived from MDSCs in serum and spleen, including hydrogen dioxide （H202） and nitric oxide （NO）, and the activity of nitric oxide synthase （NOS） were determined during the mobilization. Apop- tosis of T lymphocytes was assessed by using Annexin-V/PI. During stem cell mobilization, the number of lymphocytes and white blood cells in the peripheral blood was increased, and peaked on the 4th day. The number of stem cells in G-CSF-treated mice was significantly greater than that in controls （P〈0.01）. The expansions of MSDCs were also observed after G-CSF mobilization, with a more notable rate of growth in the peripheral blood than in the spleen. The activity of NOS and the production of NO were increased in the donor mice, and the serum H202 levels were approximately 4-fold greater than the con- trois. Consequently, apoptosis of T lymphocytes was increased and showed a positive correlation with the elevated percentage of MDSCs. It was concluded that G-CSF could provide sufficient peripheral blood stem cells for transplantation. Exogenous administration of G-CSF caused the accumulation of MDSCs in the peripheral blood and the spleen, which could lead to apoptosis ofT lymphocytes and may offer a new strategy for the prevention and treatment of graft versus host disease.     

Effects of notch-1 down-regulation on malignant behaviors of breast cancer stem cells

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells （BCSCs）. BCSCs were enriched by using serum-free me- dium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction （RT-PCR） and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and fl0w cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a prom- ising therapeutical approach for breast cancer.     

Adenoviral transduction of PTEN induces apoptosis of cultured hepatic stellate cells

Hepatic fibrosis is the liver＇s wound healing response to virtually all forms of chronic liver injury： toxic insult, viral infection, immunological conditions and metabolic diseases. Uncontrolled liver fibrosis eventually results in cirrhosis and associated complications, such as cancer and liver failure. Given the enormous public health implications, recent improvements in the treatment of chronic liver disease have accelerated interest in uncovering the mechanisms underlying hepatic fibrosis and its resolution. Currently, it is believed that activation of quiescent hepatic stellate cells （HSC） into proliferative, contractile, and fibrogenic cells in response to liver injury appears to be the dominant driving force in fibrosis.     

Telmisartan promotes proliferation and differentiation of endothelial progenitor cells via activation of Akt

Background Numerous studies have demonstrated that the peroxisome proliferator-activated receptor-y （PPARy） plays an important role in regulating endothelial progenitor cells （EPC） function.Telmisartan,as a partial agonist of PPARy,may have an effect on the regulation of EPC functions.The purpose of this study was to investigate the effects of telmisartan on EPC proliferation and differentiation.Methods Peripheral blood derived mononuclear cells containing EPC were isolated from healthy volunteers and then cultured on flbronectin-coated dishes in the presence or absence of telmisartan.The proliferative activity of EPC was determined by colony forming units （CFU） and MTT assay.The migratory activity of EPC was assessed by transwell assay.The expression of endothelial cells （EC） markers,including vascular endothelial cadherin （VE-cadherin）,yon Willebrand factor （vWF） and endothelial nitric oxide synthase （eNOS）,were measured by Western blotting analysis.Results Morphological analysis revealed that telmisartan significantly increased the proliferation of EPC and the number of endothelial cell colony forming units.Telmisartan could enhance the expression of the makers of mature EC,including VE-cadherin,vWF,and eNOS,which indicated telmisartan could stimulate EPC to differentiate into mature EC.Telmisartan increased the phosphorylation of Akt in EPC.The inhibition of Akt activation significantly attenuated the effect of telmisartan on EPC functions,suggesting that Akt is involved in the stimulatory effect of telmisartan on EPC differentiation.Conclusions The results of this study demonstrate that telmisartan promotes EPC functions via activation of Akt.     

Preparation of PLLA/bpV（pic） Microspheres and Their Effect on Nerve Cells

In this study, we prepared PLLA/bpV（pic） microspheres, a bpV（pic） controlled release system and examined their ability to protect nerve cells and promote axonal growth. PLLA microspheres were prepared by employing the o/w single emulsification-evaporation technique. Neural stem cells and dorsal root ganglia were divided into 3 groups in terms of the treatment they received： a routine medium group（cultured in DMEM）, a PLLA microsphere group（DMEM containing PLLA microspheres alone） and a PLLA/bpV（pic） group [DMEM containing PLLA/bpV（pic） microspheres]. The effects of PLLA/bpV（pic） microspheres were evaluated by the live-dead test and measurement of axonal length. Our results showed that PLLA/bpV（pic） granulation rate was（88.2±5.6）%; particle size was（16.8±3.1）%, drug loading was（4.05±0.3）%; encapsulation efficiency was（48.5±1.8）%. The release time lasted for 30 days. In PLLA/bpV（pic） microsphere group, the cell survival rate was（95.2 ±4.77）%, and the length of dorsal root ganglion（DRG） was 718±95 μm, which were all significantly greater than those in ordinary routine medium group and PLLA microsphere group. This preliminary test results showed the PLLA/bpV（pic） microspheres were successfully prepared and they could promote the survival and growth of neural cells in DRG.     

Gastromegaly infiltrated with plasma cells: a new feature of organomegaly in patients with POEMS syndrome

Organomegaly is a major component of POEMS syndrome （an acronym of polyneuropathy,organomegaly, endocrinopatfiy, M protein, and-skin changes）, which is a rare multisystem disorder of unknown pathogenesis, In patients with POEMS syndrome, the organs involved in organomegaly usually are the liver, spleen, and lymph nodes,a-4 Three studies have reported the frequencies of hepatomegaly as 25%, 68%, and 78%; of splenomegaly as 22%, 52%, and 35%;