BACKGROUND:Previous experiments have shown that improved platelet-rich plasma can promote the proliferation of human dental pulp cells in a concentration-dependent manner, particularly when the concentration is 10%.
OBJECTIVE:To investigate the effect of platelet-rich plasma at different concentrations on the proliferation and immunogenicity of human bone marrow mesenchymal stem cells.
METHODS:Human bone marrow mesenchymal stem cells from healthy donors were cultured and passaged for 3-4 passages, identified by flow cytometry and differentiation inductions. Platelet-rich plasma samples which were manufactured from the venous blood of the same donor were used for culturing human bone marrow mesenchymal stem cells. The proliferation of human bone marrow mesenchymal stem cells was measured by cellcounting kit-8 method and the growth curves were drawn. The most suitable concentration of platelet-rich plasma was selected to culture human bone marrow mesenchymal stem cells for three generations and the Stro-1 expression rate on the surface of human bone marrow mesenchymal stem cells was determined through flow cytometry.
RESULTS AND CONCLUSION:Platelet-rich plasma at the concentration of 5%-10%evidently promoted the proliferation of human bone marrow mesenchymal stem cells on the 6th and 8th days. The most effective concentration to promote the proliferation was 10%. Platelet-rich plasma at the concentration of 10%stil promoted the proliferation of human bone marrow mesenchymal stem cells on the 10th day, and maintained a better immunogenicity of human bone marrow mesenchymal stem cells compared to the control group. These findings indicate that platelet-rich plasma can promote the proliferation of human bone marrow mesenchymal stem cells in a concentration-dependent manner, and 10%platelet-rich plasma is better to maintain the immunogenicity of human bone marrow mesenchymal stem cells.

Human adenovirus (HAdV) is one of the most important pathogens in infants and young children with acute respiratory infections and other diseases. This article reviews the literature on HAdV, including its molecular biological characteristics, detection and typing, and pathogenic mechanism, the clinical features and epidemiological characteristics of HAdV-related diseases, and the prevention and control of HAdV infections. So far, 67 types of HAdV have been identified, including recombinant variants discovered in recent years. The major epidemic strains that cause acute respiratory infections are HAdV-3 and HAdV-7, both of which belong to the subgroup B. HAdV often leads to acute respiratory infections, but it also causes diseases of other systems. HAdV-related diseases have similar clinical manifestations as those caused by other respiratory viruses, but often accompanied by gastrointestinal symptoms. The pathogenic mechanism of HAdV remains unclear, especially for the new recombinant variants, due to few studies on their association with diseases. Because there are no prospective, large randomized controlled trials of HAdV infections, the treatment of HAdV infections is controversial. Vaccine is the most effective measure to reduce respiratory HAdV infections, but it is still not commercially available.
Adenovirus Infections, Human ; virology ; Adenoviruses, Human ; classification ; genetics ; isolation & purification ; physiology ; Animals ; Humans

To explore the anti-atherosclerotic mechanism of estrogen and especially observe the effect of estradiol on the content of cholesterol in J774a.1 mouse mononuclear/macrophage-derived foam cells which were incubated with oxidized low-density lipoproteins (ox-LDL). J774a.1 mouse mononuclear/macrophages were incubated with ox-LDL or with both ox-LDL and estradiol (1, 0.1 or 0.01 micromol x L(-1)). Oil red O staining was used to observe the formation of foam cells, and cholesterol oxidase fluorometric was used to determine the content of cellular cholesterol content. Western blotting and RTFQ-PCR were used to observe the expressions of scavenger receptor class B type I (SR-B I ) in J774a.1 foam cells. Compared with the control cells, J774a.1 mouse mononuclear/macrophage-derived foam cells showed significantly increased contents of total cholesterol and cholesterol ester (P < 0.001) and decreased SR-B I mRNA expression (P < 0.01). Estradiol treatment significantly lowered the contents of total cholesterol and cholesterol ester (P < 0.05), and increased SR-B I protein and mRNA expression (P < 0.01) in the foam cells in a dose-dependent manner. Estradiol can inhibit the formation of mononuclear/macrophage-derived foam cells by decreasing the contents of total cholesterol and cholesterol ester and up-regulating the expression of SR-B I in the foam cells.
Animals ; Cell Line ; Cholesterol ; metabolism ; Cholesterol Esters ; metabolism ; Estradiol ; pharmacology ; Foam Cells ; cytology ; metabolism ; Lipoproteins, LDL ; metabolism ; Macrophages ; drug effects ; metabolism ; Mice ; Scavenger Receptors, Class B ; metabolism

Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.

Objective: To observe the clinical effect of Tongfu granules(通腑颗粒) on the gastro- intestinal dysfunction in patients with multiple organ dysfunction syndrome(MODS).Methods: The trial was prospective,multi-centric and clinically controlled.One hundred and forty patients with MODS who had been selected were randomly divided into two groups: mosapride citrate group and Tongfu granules group.Respectively at 0 hour,the 48 th hour,the 7 th day or before death, the following scoring systems were calculated: the intestinal dysfunction score,acute physiology and chronic health evaluationⅡ((APACHEⅡ)) score and Marshall score.The duration of mechanical ventilation,hospitalization in intensive care unit(ICU) and the prognosis within 28 days were recorded.Results: After treatments,the intestinal dysfunction score,(APACHEⅡ) score and Marshall score of all patients decreased,at the same time,the therapeutic effects of Tongfu granules group were more significant than those in mosapride citrate group(P20 scores.The mortality was elevated with the increased number of dysfunction organs.Conclusion: Tongfu granules can ameliorate the severity of the disease situation andimprove the prognosis of patients with MODS.

Objective To design a novel electronic device for measuring the pressure in the cuff of the artificial airway; and to study the advantage of this device on continuous and intermittent cuff pressure monitoring. Methods ① a portable electronic device for cuff pressure measurement was invented, which could turn pressure signal into electrical signal through a pressure transducer. Meantime, it was possible to avoid pressure leak from the joint and the inside of the apparatus by modified Luer taper and sophisticated design. If the cuff pressure was out of the normal range, the apparatus could release a sound and light alarm. ② Six traditional mechanical manometers were used to determine the cuff pressure in 6 tracheal tubes. The cuff pressure was maintain at 30 cmH2O (1 cmH2O =0.098 kPa) by the manometer first, and repeated every 30 seconds for 4 times. ③ Study of continuous cuff pressure monitoring: We used a random number generator to randomize 6 tracheal tubes, 6 mechanical manometers and 6 our products by number 1-6, which has the same number of a group. Every group was further randomized into two balanced groups, one group used the mechanical manometer first, and the other used our product first. The baseline pressure was 30 cmH2O, measurement was performed every 4 hours for 6 times. Results When traditional mechanical manometer was used for cuff pressure monitoring, cuff pressure was decreased by an average of 2.9 cmH2O for each measurement (F = 728.2, P = 0.000). In study of continually monitoring, at each monitoring point, the pressure measured by electronic manometer was higher than the mechanical manometer. All the pressures measured by mechanical manometer were dropped below 20 cmH2O at 8th hour, and there was no pressure decrease below 20 cmH2O measured by electronic manometer in 24 hours by contrast. In study of intermittent monitoring, the same result was found. The pressure was dropped significantly with time when measured by mechanical manometer (F = 61.795, P = 0.000), the drops below 20 cmH2O began at 8th hour; but when measured by electronic manometer, all the value stayed unchanged around the baseline in 24 hours (F = 0.511, P = 0.796). Conclusions Compared with traditional mechanical manometer, cuff pressures monitored by our novel electronic manometer were steadier in both continuous and intermittent monitoring. The device is compact and convenient, and can provide a good solution for continuously monitor of the tracheal cuff pressure.

Objective To explore the HLA-A2 restriction and immunogenicity of 5 previously identified HCV-speeific CTL epitopes. Methods Based on T2 cell, to explore the HLA-A2 restriction of previously identified HCV-specific CTL epitopes by MHC-peptide complex stabilization assay;To detect pep-tide-specific CTL in HLA-A2~+ PBMC stimulated by HLA-A2-restricted peptides by intracellular cytokine staining(ICS) and ELISPOT; To explore the cytotoxicity of peptide-specific CTL to same peptide-loaded T2 cells (target cells) by CTL cytotoxicity test. Results Among 5 previously identified CTL epitopes NS4b_78 (SMMAFSAAL) and NS5a_367 (TVSSALAEL) have high-affinity for HLA-A2 molecules(FI 1) ;ELISPOT results shown that NS4b_78(SMMAFSAAL) and NSSa_367(TVSSALAEL) induced high levels of IFN-γ-se-creting cells [(60±6) SFC/10~4 PBMC vs (4±1 ) SFC/10~4 PBMC, P < 0.01 ; (10 ± 3 ) SFC/10~4 PBMC vs (2±1 ) SFC/10~4 PBMC, P <0.01, respectively] ;ICS results indicated that there were high percentages of CD8~+ IFN-γ~+ T cells in total CD8~+T cells stimulated by these peptides [(2.33 ±0.22 ) % vs (0.05±0.01)%, P <0.001 ; (0.36±0.06)% vs (0.03±0.01)%, P <0.001, respectively]. Furthermore,peptide-specific CTL could effectively kill same peptide-loadcd T2 cells. Conclusion NS4b_78 (SMMAF-SAAL) and NSSa_367 (TVSSALAEL) were identified as HLA-A2-restricted CTL epitopes which could in-duce immune response in vitro.

Objective To identify human leucocyte antigen (HLA)-A* 0201-restricted hepatitis C virus (HCV)-cytotoxic T lymphocyte (CTL) epitopes. Methods Based on the prediction results of RANKpep and SYFPEITHI prediction programs, six candidate CTL epitopes were selected and synthesized. The affinity of candidate CTL epitopes to HLA-A* 0201 molecules of T2 cells was explored. Subsequently, enzyme-linked immunosorbent spot (ELISPOT) assay and intracellular cytokine staining (ICS) were utilized to determine whether candidate CTL epitopes could induce the recall positive response in peripheral blood mononuclear cells (PBMC) of HLA-A* 0201 positive HCV-1b-infected patients. Results Among six candidate CTL epitopes, peptides C_181(LLSCLTTPV) and NS2_172 (VLQAGLIRV) had high affinity to HLA-A* 0201 molecules. Moreover, the affinity was proportional to the concentration of peptide. Furthermore, among ten HLA-A* 0201 positive HCV-1b-infected patients, the frequencies of C_181 and NS2_172-specific interferon (IFN)-γ-producing cells were 0-19 spots forming cells (SFC)/1 × 105 PBMC and 0-20 SFC/1 × 105 PBMC, respectively.The percentages of C_ 181 and NS2_172-specific IFN-γ+ CD8+ T lymphocytes in total CD8+ T lymphocytes were 0.006%-0.065% and 0.005%-0.080%, respectively. Conclusion Peptides C_181 (LLSCLTTPV) and NS2_172 (VLQAGLIRV) are identified as novel HLA-A* 0201-restricted HCV-CTL epitopes.

Objective To compare the therapeutical effect of puerarin, ligustrazine, ginsenoside Rb1, Hydroxysafflor yellow A on cerebral ischemia reperfusion mice. Methods The mice were randomly assigned for sham group, model group, puerarin group, ligustrazine group, ginsenoside Rb1 group, and Hydroxysafflor yellow A group, 24 mice for each group. All the groups were subjected to middle cerebral artery occlusion (MCAO) by 1 h ischemia and 24 h of reperfusion except the sham group. The puerarin, ligustrazine, ginsenoside Rb1, Hydroxysafflor yellow A were administrated by tail vein injection with 3μmol/kg at the onset of 1 h of ischemia. The neurologic deficit score, infarct area calculated by TTC staining, cerebral cortex blood flow monitored by laser doppler flowmetry, NO content measured by chemical colorimetry and western blot were applied to determine the expression for cleaved-caspase-3 and nuclear transcription factor NF-κB for each group. Results Compared with the model group, the infarct area (15.83%± 1.83%, 22.00%± 2.53%, 22.83%± 1.83%, 17.83%± 1.72%vs. 34.67%± 2.66%) in the puerarin group, ligustrazine group, ginsenoside Rb1 group, Hydroxysafflor yellow A group was significantly decreased (P<0.01 or P<0.05);the cerebral cortex blood flow (598.81 ± 9.90 μl/kg?min-1, 614.78 ± 9.20 μl/kg?min-1, 577.83 ± 5.55 μl/kg?min-1, 583.54 ± 7.98 μl/kg?min-1 vs. 548.43 ± 1.97 μl/kg?min-1) significantly increased (P<0.01 or P<0.05);the NO content (17.09 ± 1.18μmol/L, 18.54 ± 0.54μmol/L, 18.17 ± 0.49μmol/L, 15.10 ± 0.73μmol/L vs. 20.63 ± 0.73μmol/L) ignificantly decreased (P<0.01 or P<0.05);the expression of cleaved-caspase-3 (1.02 ± 0.08, 1.12 ± 0.04, 0.87 ± 0.08, 1.07 ± 0.08 vs. 1.30 ± 0.06) and NF-κB p-p65/NF-κB p65 (1.03 ± 0.19, 1.15 ± 0.05, 1.12 ± 0.08, 0.72 ± 0.08 vs. 1.45 ± 0.08) ignificantly decreased (P<0.01 or P<0.05) Conclusions Four Chinese herbal monomers could improve nerve and cerebral dysfunctions and ameliorate ischemia symptoms with varying degrees. The mechanisms were involved with the enhancement of cerebral cortex blood flow and inhibition of cell apoptosis and the activation of inflammatory signaling pathways.

Objective:To investigate the effect of modified plateletrich plasma(mPRP)on the osteogenic differentiation of stemcells from human exfoliated deciduous teeth(SHED).Methods:mPRP at 1%,2%,5%,10% and FBS at 10% were added to thecultured SHED of passage 4,respectively.The influence of mPRP on alkaline phosphatase(ALP)activity was evaluated using ALPkit.RUNX2 and osteocalcin mRNA expression in the treated cells were examined by realtime PCR.Results:mPRP enhanced ALPactivity in the SHED,and the effect of mPRP was more obvious at 2%.Treatment of the cells with 2% mPRP upregulated the mRNAexpressions of RUNX2 and osteocalcin.Conclusion:mPRP can promote the osteogenic differentiation of SHED.