Objective This study investigated the regulatory effect of high mobility group protein B1 (HMGB1) in the peripheral blood of patients with primary immune thrombocytopenia (ITP) on myeloid dendritic cells (mDC) and Th17/regulatory T cells (Treg) balance. Methods The study enrolled 30 newly diagnosed ITP patients and 30 healthy controls.Flow cytometry was used to measure the proportion of mDC, Th17, and Treg cells in the peripheral blood of ITP patients and healthy controls. ELISA was conducted to quantify the serum levels of HMGB1, interleukin 6 (IL-6), IL-23, IL-17, and transforming growth factor β(TGF-β). The mRNA levels of retinoic acid-related orphan receptor γt(RORγt) and forehead box P3(FOXP3) were detected by real-time PCR. The correlation between the abovementioned cells, cytokines, and platelet count was assessed using Pearson linear correlation analysis. Results The proportion of Th17 cells and the expression levels of HMGB1, IL-6, IL-23, IL-17 and the level of RORγt mRNA in the peripheral blood of ITP patients were higher than those in healthy controls. However, the Treg cell proportion and TGF-β level were lower in ITP patients than those in healthy controls. In patients with ITP, the proportion of mDC and the level of FOXP3 mRNA did not show significant changes. The proportion of mDC cells was significantly correlated with the expression of IL-6 and IL-23. Moreover, the expression of HMGB1 showed a significant correlation with the expression of mDC, IL-6, IL-23, RORγt mRNA, and IL-17. Notably, both the proportion of mDC cells and the expression of HMGB1 were negatively correlated with platelet count. Conclusion The high expression of HMGB1 in peripheral blood of ITP patients may induce Th17/Treg imbalance by promoting the maturation of mDC and affecting the secretion of cytokines, thereby potentially playing a role in the immunological mechanism of ITP.
Humans ; Th17 Cells/cytology* ; HMGB1 Protein/genetics* ; T-Lymphocytes, Regulatory/cytology* ; Female ; Male ; Dendritic Cells/metabolism* ; Adult ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic/genetics* ; Nuclear Receptor Subfamily 1, Group F, Member 3/genetics* ; Young Adult ; Interleukin-23/blood* ; Interleukin-17/blood* ; Interleukin-6/blood* ; Forkhead Transcription Factors/genetics* ; Myeloid Cells/cytology* ; Aged

Objective:To investigate the role of autophagy mediated by mTOR signaling pathway in the inhibition of osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) induced by cadmium.Methods:HBMSCs were divided into 0, 2.5 or 5.0 μmol/L groups according to the exposure dose of cadmium chloride (CdCl 2), and each group was treated for 1 day, 4 days and (or) 7 days. The ALP activity and mRNA and protein expression levels of osteogenesis markers (ALP, RUNX2 and OSTERIX), autophagy-related proteins (LC3 and Beclin-1) and mTOR signaling pathway related proteins (mTOR, p-mTOR and p-p70S6K) expression, alkaline phosphatase staining and alizarin red staining were detected. MHY 1485 was selected as the signaling pathway activator. The control group, CdCl 2 group (5.0 μmol/L), MHY 1485 group and CdCl 2+MHY 1485 combined treatment group were set. After 7 days of treatment, the expression levels of autophagy related proteins and mTOR signaling pathway related proteins of hBMSCs in each group were detected. Results:There was no significant difference in ALP activity between 0, 2.5 and 5.0 μmol/L groups on day 1 and 4 ( P>0.05); On day 7, compared with the 0 μmol/L group, the ALP activity, expression of osteogenic markers (ALP, RUNX2, OSTERIX) and mTOR signaling pathway related proteins (mTOR, p-mTOR, p-p70S6K) expression decreased in the 2.5 and 5.0 μmol/L group ( P<0.05). Compared with the 0 μmol/L group, the staining of the 2.5 and 5.0 μmol/L groups became lighter, and the formation of ALP and mineralized nodules was reduced. Compared with the CdCl 2 group, the autophagy related protein expression in the CdCl 2+MHY 1485 combined treatment group decreased, and the mTOR signaling pathway related protein expression increased. The difference was statistically significant ( P<0.05). Conclusion:The inhibition of osteogenic differentiation of hBMSCs by cadmium may be related to autophagy mediated by mTOR signaling pathway.

Objective:To investigate the role of autophagy mediated by mTOR signaling pathway in the inhibition of osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) induced by cadmium.Methods:HBMSCs were divided into 0, 2.5 or 5.0 μmol/L groups according to the exposure dose of cadmium chloride (CdCl 2), and each group was treated for 1 day, 4 days and (or) 7 days. The ALP activity and mRNA and protein expression levels of osteogenesis markers (ALP, RUNX2 and OSTERIX), autophagy-related proteins (LC3 and Beclin-1) and mTOR signaling pathway related proteins (mTOR, p-mTOR and p-p70S6K) expression, alkaline phosphatase staining and alizarin red staining were detected. MHY 1485 was selected as the signaling pathway activator. The control group, CdCl 2 group (5.0 μmol/L), MHY 1485 group and CdCl 2+MHY 1485 combined treatment group were set. After 7 days of treatment, the expression levels of autophagy related proteins and mTOR signaling pathway related proteins of hBMSCs in each group were detected. Results:There was no significant difference in ALP activity between 0, 2.5 and 5.0 μmol/L groups on day 1 and 4 ( P>0.05); On day 7, compared with the 0 μmol/L group, the ALP activity, expression of osteogenic markers (ALP, RUNX2, OSTERIX) and mTOR signaling pathway related proteins (mTOR, p-mTOR, p-p70S6K) expression decreased in the 2.5 and 5.0 μmol/L group ( P<0.05). Compared with the 0 μmol/L group, the staining of the 2.5 and 5.0 μmol/L groups became lighter, and the formation of ALP and mineralized nodules was reduced. Compared with the CdCl 2 group, the autophagy related protein expression in the CdCl 2+MHY 1485 combined treatment group decreased, and the mTOR signaling pathway related protein expression increased. The difference was statistically significant ( P<0.05). Conclusion:The inhibition of osteogenic differentiation of hBMSCs by cadmium may be related to autophagy mediated by mTOR signaling pathway.

Objective To investigate the expression of microRNA-21(miR-21)in breast cancer cell lines and serum of patients with breast cancer and the impact on the invasion and migration of breast cancer cells.Methods From Jan 2013 to Feb 2014, miR-21 expression were determined by fluorescent quantity polymerase chain reaction (FQ-PCR) in 4 breast cell lines (HBL-100, MCF-7, MDA-MB-231 and MDA-MB-468) and in serum from breast cancer patients ( n =56 ) , breast benign disease patients ( n =39 ) andhealth controls ( n =45 ) . The characteristics of cell invasion and migration were examined by transwellinvasion and migration assay afterbreast cancer cell line MDA-MB-231 were transfectedwith miR-21 inhibitor or negative control by lipofectamin.The t test was used to analysis the normal distribution data. Results FQ-PCR results showed that the relative expression of miR-21 in the normal breast epithelial cell line HBL-100 was 1.01 ±0.04, in the breast cancer cell line MCF-7, MDA-MB-231 and MDA-MB-468 were 1.99 ±0.11,4.02 ±0.38 and 3.73 ±0.79 respectively.Compared with the normal controls, miR-21 were highly expressed in the three breast cancer cell lines, the difference was statistically significant (t=9.01, 9.20 and 4.55, respectively, P<0.01); and the miR-21 was highly expressed in invasive and metastatic breast cancer cell lines (MDA-MB-231 and MDA-MB-468),compared with weakly invasive breast cancer cell line MCF-7, the difference was statistically significant ( t values were 6.14 and 2.91, P<0. 05), suggesting that miR-21 is highly expressed in breast cancer cells, and is closely related to the invasion and metastasis.The relative expression of miR-21 in serum of breast cancer was 2.63 (1.57-4.59), in benign breast disease group was 1.34 (1.01-1.78), in healthy control group was 0.81 (0.52-1.59), the miR-21 expression in the serum of breast cancer patients was significantly higher than in patients with benign lesions and normal control group (U values were 208 and 279, P<0.01), whereas no significant difference in serum in patients with benign lesions and normal control group, the miR-21 expression in the serum of breast cancer patients with lymph node metastasis (U=95 , P=0.19) was 3.55 (2.44-5.26), significantly higher than those without lymph node metastasis [2.11(1.59-3.25), U=216,P=0.021]. The results of invasion and migration assay showed that cells treated with miR-21 inhibitor invasion was:44 ±18, the number of cell migration was:98 ±22, while the negative control treated cells after invasion was:133 ±44, migration cell number:255 ±35;miR-21 inhibitor treatment compared with the negative control, cell invasion and migration was also significantly decreased( t values were 5.46 and 9.08, P<0. 01) .The cell invasion and migration assay indicated the numbers of MDA-MB-231 cells, which invaded or migrated to lower chamber, were 44 ±18 and 98 ±22 respectively after miR-21 inhibitor was applied, The numbers of invaded or migrated cells were 133 ±44 and 255 ±35 when the negative control was applied.The ability of cell invasion and migration was decreased significantly in the inhibitor group compared with the negative group(tvalue separately was 5.46, 9.08, P<0.01).The capacity of breast cancer cell invasion and migrationwas significantly decreased after transfection ofmiR-21 inhibitor.Conclusions MiR-21 is highly expressed in breast cancer cell lines and breast cancer patients′serum.Altered expression of miR-21 maybeplays an important role in breast cancer invasion and migration.MiR-21 may serve as new biomarker to early detectionand prognosis estimation of breast cancer.

Objective To investigate the expression level of microRNA-145 in breast cancer cell lines andtissues and its impact on breast cancer invasion and metastasis.Methods MiR-145 expression was detected by FQ-PCR in 5 breast cancer cell lines ( HBL-100, MCF-7, MDA-MB-231, MDA-MB-468 and SK-BR-3)and in breast cancer tissue and paraneoplastic tissues (n=39).The miR-145 expression plasmid ( Psif-miR-145 ) and negative control plasmid were transfected into SK-BR-3 using lipofectamine, respectively.The characteristics of invasion and migration of the transfected SK-BR-3 cells were examined by scratch test and transwell assay.The target genes of miR-145 were predicted by bioinformatics and the ANGPT2 gene were verified as miR-145 target by the dual-luciferase reporter assay.The expression levels of ANGPT2 protein was examined by western blot after pSIF-miR-145 transfection by lipofectamine in breast cancer cell line SK-BR-3.Results FQ-PCR result indicated that miR-145 expression level waslower in breast cancer tissue (45.93 ±22.02)than paraneoplastic tissue [ (182.04 ±56.92), U value was 7, P<0.01].MiR-145 expression level was lower in breast cancer cell lines than normal breast cells.miR-145expression in 4 breast cancer cell lines was 0.51 ±0.05, 0.07 ±0.01, 0.36 ±0.04 and 0.04 ±0.01, respectively.Compare with normal breast cell, miR-145 was lower expressed in all 4 breast cancer cell lines (t value separately was 15.93, 308.17, 25.02, 201.30;P<0.05).Lower expression of miR-145 was observed in the highly invasive breast cancer cells (MDA-MB-231, MDA-MB-468 and SK-BR-3), compared with weakly invasive breast cancer cell (MCF-7) (t value separately was14.18, 3.78, 15.20;P<0.05). Wound healing assay shows that overexpression of miR-145 in SK-BR-3 significantly reducedthe motility as compared with control group (P <0.01).The cell invasion assay indicated the numbers of miR-145 overexpressed SK-BR-3 cells, which invased to lower chamber, was 137 ±37, the numbers of invased cells was 617 ±80 when the negative control was applied. Over-expression of miR-145 could repress the expression levelsof ANGPT2 protein;miR-145 could repress the activity of luciferase reporter carrying a 3′-untranslated region of ANGPT2 mutated the predicted binding site, the activity of luciferase was reversed. Conclusions MiR-145 depressed in breast cancer cell lines and breast cancer tissues.MiR-145 maybe plays an important role in breast cancer invasion and migration by directly target ANGPT2.

Objective To study the facial cosmetic zonation and its application of adipose injection for facial juvenescence.Methods According to standard of aesthetics,we divided total face into ten cosmetic units and 49 cosmetic sub-units.23 patients,injected adipose for facial juvenescence,were evaluated.Results 149 cosmetic units from 23 patients were injected with adipose granules,and involved 300 sub-units.Each unit was injected with adipose from 1 to 35 ml.After injection,the faces showed swelling in varying degrees,which resolved from ten to fourteen days.Local ecchymosis occurred in 11 cases,and resolved from five to twenty days.No any complication appeared,such as infection,scleroma and vascular embolism and so on.The follow-up results showed that one month later post-operation,swelling on injective sites resolved,and appearances were stable.3-6 months later post-operation,the injective sites were fuller.One year later,the good j uvenescence effect achieved,and the patients felt satisfactory.Conclusions On the basis of facial cosmetic zonation,the whole cosmetic evaluation combined with local cosmetic evaluation might be helpful to carry out adipose injection for facial juvenescence,and reach a satisfactory result.

Objective To prepare the virus-like particles (VLPs) of Chikungunya virus (CHIKV) and evaluate the immunogenicity.Method CHIKV structural protein C-E3-E2-6K-E1 encoding gene were amplified by fusion PCR,and cloned into an insect cell expression vector.The recombinant Baculovirus were recovered by co-transfection of the expression plasmid with baculovirus linear DNA into SF9 insect cells,and CHIKV VLPs were prepared from suspension culture SF9 cells.Structural proteins expression were analyzed using IFA,SDS-PAGE and Western-Blot,and morphological analysis via electron microscopy.Result Which showed that CHIKV structural proteins were secreted into the cell culture supernatant and assembled into virus-like spherical particles.BALB /c mice were immunized with the VLPs,high levels of CHIKV specific antibodies were detected in the sera,and CHIKV infection of Vero cells could be effectively neutralized.Couclusion The results showed that CHIKV VLPs can be efficiently produced by the baculovirus expression system in insect cells,and specific IgG and neutralization antibodies could be induced in mice after VLPs immunization.This research laid the foundation for the development of CHIKV VLPs based on immunological detection reagents and even vaccines.

Objective To investigate the dose-response pattern on the inducible expression of PIG3 mRNA in normal human lymphoblastoid AHHI cells by 60Co γ-rays,and its possibility for developing novel radiation biodosimeter.Methods Laser confocal fluorescent microscopy was used to detect the γ-H2AX foci,a biomarker of DNA double-strand break.After irradiation with 0,1,2,4,6,8 and 10 Gy of 60Coγ- rays,AHH-1 cells were harvested at 4,10 and 24 h post-irradiation,and subjected to total RNA extraction and detection of PIG3 mRNA expression by quantitative real-time PCR.Results PIG3 protein foci were formed in the nuclei at 30 min after irradiation,and a part of these PIG3 foci were colocalized with γH2AX foci.After irradiation,PIG3 mRNA level was enhanced with the increased time of postirradiation,and remained an increased level at least till 24 h.The radiation-inducible expression of PIG3 mRNA was demonstrated in a dose-dependent manner.The dose-dependent range at 4 h post-irradiation was 0 - 6 Gy,and at 10 h and 24 h was 0 - 10 Gy.Conclusions PIG3 involves in the cellular response to DNA double-strand break.The dose-dependent inducible response of PIG3 mRNA expression might provide a valuable candidate for developing a novel radiation biodosimeter.

Objective To evaluate the radioprotective effect of vanillin derivative VND3207 on DNA damage induced by different LET ionizing radiation.Methods The plasmid DNA in liquid was irradiated by 60Co γ-rays, proton or 7Li heavy ion with or without VND3207.The conformation changes of plasmid DNA were assessed by agarose gel electrophoresis and the quantification was done using gel imaging system.Results The DNA damage induced by proton and 7Li heavy ion was much more serious as compared with that by 60Co γ-rays, and the vanillin derivative VND3207 could efficiently decrease the DNA damage induced by all three types of irradiation sources, which was expressed as a significantly reduced ratio of open circular form (OC) of plasmid DNA.The radioprotective effect of VND3207 increased with the increasing of drug concentration.The protective efficiencies of 200 μmol/L VND3207 were 85.3% (t =3.70,P =0.033), 73.3% (t = 10.58, P =0.017)and 80.4% (t =8.57,P =0.008)on DNA damage induction by 50 Gy of γ-rays, proton and 7Li heavy ion, respectively.It seemed that the radioprotection of VND3207 was more effective on DNA damage induced by high LET heavy ion than that by proton.Conclusions VND3207 has a protective effect against the genotoxicity of different LET ionizing radiation, especially for γ-rays and 7 Li heavy ion.

Emergency in public health is one major issue relating to the national economy and the people's livelihood.In dealing with the key technologies of detects,diagnosis,disinfection,prevention and control,it is wise to syncretize the correlated subjects in nucleus,chemistry and biology,improve the support condition of teaching,strengthen the reform of teaching contents,add some lessons of emergency treatment,make emergency equipments,and form the ability of rapid diagnosis on the spot and precise identifying in lab,for cultivating the students'capability of spot handling,organization and command,and furthermore exploring and practicing the training pattern of the public heath talent with high quality and emergency ability in our country.