OBJECTIVETo investigate the expression of chemokine stromal cell-derived factor-1 (SDF-1) receptor CXCR4 in human gingival mesenchymal stem cells (GMSCs) and the migration potential of GMSCs stimulated with SDF-1.

METHODSHuman GMSCs were isolated by single-cell cloning method. Their cell surface markers were characterized by flow cytometry, and the rate of colony formation was evaluated. Differentiation assay was used to detect the differentiation potential of GMSCs. The expression of chemokine SDF-l receptor CXCR4 in GMSCs was detected by immunocytochemical staining. The chemotactic effect of SDF-1 on GMSCs was detected using a 24-multiwell Transwell cell culture chamber. The number of net migrated cells was counted in different microscope fields.

RESULTSHuman GMSCs possessed high self-renewal potential and formed single-cell colonies cultured in vitro. GMSCs expressed mesenchymal stem cells-associated markers CD44, CD73, CD90, CD105, and CD166, and the expression of hemopoietic stem cell surface markers CD14, CD34, and CD45 was negative. GMSCs differentiated into osteoblasts and adipocytes under defined culture conditions. The colony forming unit-fibroblastic for GMSCs was 21.4%/±2.8%. Immunocytochemical staining demonstrated that GMSCs expressed chemokine SDF-1 receptor CXCR4. The number of GMSCs migrating at concentrations of 100 ng.mL-1 and 200 ng.mL-1 of SDF-l in the Transwell cell culture chamber was significantly higher than that of the negative control (189.3±4.4, 164.6±4.9 cells/field vs. 47.8±2.5 cells/field, P<0.01). Treatment with the CXCR4 neutralizing antibody, an antagonist for CXCR4, significantly reduced the migratory effect compared with the negative controls (29.0±2.4 cells/field vs. 47.8±2.5 cells/field, P<0.01).

CONCLUSIONHuman GMSCs express chemokine SDF-l receptor CXCR4. SDF-1 may participate in regulating chemotaxis of human GMSCs. Results suggest that the migration induced by SDF-1 is mediated by CXCR4.

Adipocytes ; Cell Culture Techniques ; Cell Differentiation ; Chemokine CXCL12 ; metabolism ; Chemotaxis ; Flow Cytometry ; Gingiva ; physiology ; Humans ; Mesenchymal Stromal Cells ; physiology ; Osteoblasts ; Receptors, CXCR4 ; Signal Transduction

This article was aimed to study macroporous resin adsorption kinetics for effective extraction of water ex-tracting with alcohol precipitating in cicada slough. PT, APTT and the coagulation-fibrinolysis dynamic figure were taken as main indexes, which were combined with static and dynamic tests, to select the best macroporous resin to separate and purify the extraction. Adsorption kinetics curve was drawn to fit the adsorption kinetics model. The re-sults showed that NKA-9 macroporous resin was more effective in separating and purifying effective extraction than others. The adsorption dynamic behavior was well described by the pseudo-second-order kinetics equation. It was concluded that the adsorption rate was mainly controlled by the intraparticle diffusion.

BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

Objective To explore the clinical and imaging factors influencing the patients' prognosis after preoperative radiotherapy for local advanced rectal cancer.Methods We retrospectively analyzed 106 locally advanced rectal cancer patients from June 2004 to September 2015 in our institution.All patients underwent preoperative radiotherapy.According to the Mandard score,patients were divided into 5 groups (TRG1-5).All patients were divided into two groups according to the TRG,which including good responder (TRG1 + 2) and poor responder (TRG3 + 4 + 5) groups.All of the tumor ADC values of post-RT were measured by Diffusion-weighted MRI technology,and the relationship between tumor ADC values of post-RT and TRG was analyzed.Results In univariate analysis,age,chemotherapy,pT,pN,differentiation degree,vascular invasion and TRG were significantly associated with overall survival (x2 =3.945-8.110,P ＜ 0.05).Multivariate analysis indicated that differentiation degree and TRG were the independent prognostic factors for OS (x2 =5.221,6.563,P ＜ 0.05).No significant difference was found between long-course and short-course radiotherapy group (P ＞ 0.05) in OS.The good responder group had a favorable survival in 5-year OS compared to the poor responder group (x2 =8.110,P ＜ 0.05).Preoperative radiotherapy,preoperative chemotherapy,pathological type,differentiation degree and gross type,vascular tumor thrombus and tumor ADC values of post-RT were significantly associated with TRG (x2 =4.189-18.139,P ＜ 0.05).The best critical point of tumor ADC values of post-RT was 1.7 x 10-3 mm2/s by using ROC curve.The accuracy of tumor ADC values of post-RT in predicting TRG1 + 2 was 70％.Conclusions The TRG can predict the efficacy of preoperative radiotherapy in patients with locally advanced rectal cancer based on the Mandard score.There was no significant difference in OS between long-course radiotherapy group and short-course radiotherapy group.The tumor ADC values of post-RT might become a potential factor to predict TRG in patients with locally advanced rectal cancer after preoperative radiotherapy.

Objective To investigate the distribution and clonality of the T-cell receptor (TCR) Vβ repertoire in chronic graft versus host disease (cGVHD).Methods The complementarity determining region 3 (CDR3) of the TCRβ gene with 24 variable regions was amplified in peripheral blood mononuclear cells drawn from one cGVHD patient after allogenic bone marrow transplantation (allo-BMT) 35, 39, 43 or 45 months respectively, using RT-PCR, to observe the expression of TCR Vβ repertoire T cells. The PCR products were further analyzed by genescan to evaluate clonality of T cells. Ressults Fourteen or 16 TCR Vβ subfamily T ceils were detected in each sample of cGVHD case. Oligoclonal T cells were identified in TCR Vβ 6, 16, 17, 19 and 21 subfamilies. The stable clonal T cells in all samples were identified in Vβ6, Vβ17 and Vβ21 subfamilies.Conclusion Skewing distribution and stable clonal expansion of T cells can be found in cGVHD cases and it may be related to the initiation of cGVHD.

OBJECTIVETo investigate the chemotactic response of human periodontal ligament stem cells (PDLSCs) to bone morphogenetic protein-2 (BMP-2).

METHODSHuman PDLSCs were obtained from clinically healthy premolars extracted for orthodontic reasons and used to isolate PDLSCs by limited dilution method. The expression of Vimentin and stem cell marker STRO-1 on PDLSCs were demonstrated with immunocytochemical staining. Differentiation assay was used to detect the differentiation potential of PDLSCs. Cloning formation experiment and 5-bromo-2-deoxyuridine (BrdU) incorporation assay were used to determine the stem cell characteristics of PDLSCs. The chemotactic effect of BMP-2 on PDLSCs was detected by using a 24-multiwell Transwell cell culture chamber. The number of net migrated cells was counted in different microscope fields.

RESULTSHuman PDLSCs displayed positive staining for Vimentin and expressed the stem cell marker STRO-1. These cells differentiated into osteoblasts and adipocytes under defined culture conditions, possessed high self-renewal potential and formed single-cell colonies in vitro. The number of cells migrating at concentrations of 100, 200 ng mL(-1) of BMP-2 in Transwell cell culture chamber was significantly higher than that of negative control (P<0.01).

CONCLUSIONBMP-2 may participate in regulating chemotaxis of human PDLSCs.

Adipocytes ; Bone Morphogenetic Protein 2 ; Cell Culture Techniques ; Cell Differentiation ; Humans ; Osteoblasts ; Periodontal Ligament ; Stem Cells

Objective To investigate the predictive value of androge receptors in stage pT1 non-muscle-invasive bladder cancer.Methods A total of 196 patients with stage pT1 non-muscle-invasive bladder cancer who underwent a transurethral resection of the bladder in Baoji People's Hospital from February 2003 to June 2012 were recruited to carry a retrospective analysis.The mRNA expression of the androge receptors transcript variants 1 (AR1) and 2 (AR2) was measured by quantitative real-time PCR.Spearman's correlation coefficient was used to analysis the correlation of androge receptors mRNA level to KRT5 and KRT20 mRNA.Results Kaplan-Meier analysis indicated that the recurrence-free survival,progression-free survival and cancer specific survival of patients with high AR1 mRNA expression (≥35.47) were significant better than patients with low AR1 mRNA expression (P ＜0.05).Multivariate COX regression analysis revealed that high AR1 mRNA expression was an independent prognostic marker for recurrence-free survival and cancer specific survival (P ＜ 0.05).Spearman rank correlation revealed a significant positive association between mRNA expression of AR1 and KRT5 (rs =0.3171,P ＜ 0.001) as well as a negative association with multifocal tumors (rs =0.1478,P ＜ 0.05).Conclusion Androge receptors mRNA expression can predict recurrence-free survival and cancer specific survival in patients with stage T1 nonmuscle-invasive bladder cancer.Further studies on the levels of androge receptors mRNA tend to be particularly important.

Objective To observe the effect of Proanthocyanidin on motor after spinal cord injury (SCI) in rats. Methods 36 healthy adult SD rats were divided into groups A, B and C (n=12), and SCI was induced with Allen's mode (250 g·mm) on T9. Proanthocyanidin 40 mg/kg was injected intraperitoneally for group A, methylprednisolone (MP) 30 mg/kg for group B and the same volume of saline for group C 30 min after SCI. 1 d, 3 d and 7 d after operation, all the rats were assessed with Basso-Beattie-Bresnahan (BBB) scale and slanting board test, and their serumal malondialdehyde (MDA) and superoxide dismutase (SOD) were detected. Results The scores of BBB and the slanting board test imporoved more in group A and group B than in group C (P<0.05). The SOD increased and MDA decreased in groups A and B significantly 1 d and 3 d after operation compared with those of group C (P<0.05), and only in group A 7 d after operation (P<0.05). Conclusion Proanthocyanidin may inhibit the lipid peroxidation and promote the recovery of motor after spinal cord injury in rats.

ObjectiveTo?investigate?the?expression?of?chemokine?stromal?cell-derived?factor-1?(SDF-1)?receptor?CXCR4?in?human?gingival?mesenchymal?stem?cells?(GMSCs)?and?the?migration?potential?of?GMSCs?stimulated?with?SDF-1.?Methods?Human?GMSCs?were?isolated?by?single-cell?cloning?method.?Their?cell?surface?markers?were?characterized?by?flow?cytometry,?and?the?rate?of?colony?formation?was?evaluated.?Differentiation?assay?was?used?to?detect?the?differentiation?potential?of?GMSCs.?The?expression?of?chemokine?SDF-1?receptor?CXCR4?in?GMSCs?was?detected?by?immunocytochemical?staining.?The?che-motactic?effect?of?SDF-1?on?GMSCs?was?detected?using?a?24-multiwell?Transwell?cell?culture?chamber.?The?number?of?net?migrated?cells?was?counted?in?different?microscope?fields.?Results???Human?GMSCs?possessed?high?self-renewal?potential?and?formed?single-cell?colonies?cultured?in vitro.?GMSCs?expressed?mesenchymal?stem?cells-associated?markers?CD44,?CD73,?CD90,?CD105,?and?CD166,?and?the?expression?of?hemopoietic?stem?cell?surface?markers?CD14,?CD34,?and?CD45?was?negative.?GMSCs?differentiated?into?osteoblasts?and?adipocytes?under?defined?culture?conditions.?The?colony?forming?unit-fibroblastic?for?GMSCs?was?21.4%±2.8%.?Immunocytochemical?staining?demonstrated?that?GMSCs?expressed?chemokine?SDF-1?receptor?CXCR4.?The?number?of?GMSCs?migrating?at?concentrations?of?100?ng·mL-1?and?200?ng·mL-1?of?SDF-1?in?the?Transwell?cell?culture?chamber?was?significantly?higher?than?that?of?the?negative?control?(189.3±4.4,?164.6±4.9?cells/field?vs.?47.8±2.5?cells/field,?P<0.01).?Treatment?with?the?CXCR4?neutralizing?antibody,?an?antagonist?for?CXCR4,?significantly?reduced?the?migratory?effect?compared?with?the?negative?con-trols?(29.0±2.4?cells/field?vs.?47.8±2.5?cells/field,?P<0.01).?Conclusion???Human?GMSCs?express?chemokine?SDF-1?receptor?CXCR4.?SDF-1?may?participate?in?regulating?chemotaxis?of?human?GMSCs.?Results?suggest?that?the?migration?induced?by?SDF-1?is?mediated?by?CXCR4.

OBJECTIVETo determine the effect of aspirin on cell proliferation, alkaline phosphatase (ALP) activity, cell cycle and apoptosis in murine bone marrow stromal cells, so as to explore an appropriate dose range to improve bone regeneration in periodontal treatment.

METHODSST2 cells were stimulated with aspirin (concentrations of 1, 10, 100 and 1 000 μmol/L) for 1, 2, 3, 5 and 7 d. Cell proliferation was measured by methyl thiazolyl tetrazolium (MTT) assay. After ST2 cells were treated for 1, 3 and 7 d, ALP activity was measured by ALP kit, cell cycle and apoptosis were measured by flow cytometry (FCM) after treated for 48 h.

RESULTSMTT assays showed that various doses of aspirin have different effects on the cell growth. Briefly, lower concentrations (1, 10 μmol/L) of aspirin promoted the cell growth, the A value of 0, 1 and 10 μmol/L aspirin 7-day-treated cells were 0.313±0.012, 0.413±0.010 and 0.387±0.017 respectively (P <0.01 vs control), and so did the ALP level ([4.3±0.9], [6.0±0.3] and [7.7±0.4] μmol·min(-1)·g(-1), P <0.05 vs control), while higher concentrations, especially 1000 μmol/L of aspirin might inhibit the cell growth with time going, A value and ALP level were 0.267±0.016, (4.3±1.3) μmol·min(-1)·g(-1) respectively (P <0.05 vs control). Cell cycle analysis revealed no changes in comparison to control cells after treatment with 1 or 10 μmol/L aspirin, but it was observed that cell mitosis from S phase to G2/M phase proceeded at higher concentrations of 100 μmol/L aspirin, and the cell cycle in phase G0/G1 arrested at 1000 μmol/L. Parallel apoptosis/necrosis studies showed that the percentage of cells in apoptosis decreased dramatically at all doses of aspirin, the apoptosis rates of ST2 cells responded to 0, 1, 10, 100 and 1000 μmol/L aspirin were (11.50±0.90)%, (5.30±0.10)%, (5.50±0.10)%, (4.90±0.90)% and (7.95±0.25)% respectively (P<0.05 vs control).

CONCLUSIONSThis study demonstrated that lower dosage of aspirin can promote ST2 cells growth, osteogenic activity and inhibit its apoptosis. Aspirin maybe used for the bone reconstruction with a proper concentration.

Alkaline Phosphatase ; metabolism ; Animals ; Apoptosis ; drug effects ; Aspirin ; administration & dosage ; pharmacology ; Bone Regeneration ; Cell Cycle ; drug effects ; Cell Division ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Formazans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; enzymology ; Mice ; Periodontics ; Tetrazolium Salts ; Time Factors