Animals ; Bone Regeneration ; Bone Substitutes ; Computer-Aided Design ; Humans ; Lactic Acid ; chemistry ; Microsurgery ; Neovascularization, Physiologic ; Polyesters ; chemistry ; Polyglycolic Acid ; chemistry ; Starch ; chemistry ; Stem Cells ; cytology ; Tissue Engineering ; methods

Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; Neovascularization, Physiologic ; Osteogenesis

The expression of paired immunoglobin-like receptors A (PIR-A) and B (PIR-B) and their relationship with tolerogenic dendritic cells (T-DC) in mice were investigated. The mouse DCs line, DC2.4 cells were cultured with the recombinant murine interleukin-10 (IL-10) and recombinant human transforming growth factor beta1 (TGF-beta1) respectively to develop the T-DC and stimulated with lipopolysaccharide (LPS) for 48 h to induce the mature dendritic cells (LPS-DC). Special small interfering RNAs (siRNA) molecule for PIR-B was chemically synthesized and transfected into DC2.4 cells (Si-DC) by lip2000. The expression of PIRs on DC2.4 cells were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), flow cytometry (FCM) and Western blot. Realtime reverse transcriptase-polymerase chain reaction (Realtime-PCR) was applied for measurement of PIR-A and CD80, CD86, MHC-II mRNA expression. The allogeneic stimulating capacity of DCs was measured by mixed lymphocyte reaction (MLR) using (3)H-thymidine incorporation test. The concentration of IFN-gamma in supernatants of MLR from distinct groups was analyzed by ELISA. The results showed that PIR positive rate was (28.65+/-8.12)% examined by FCM on DC2.4 cells. PIR positive rate was increased dramatically to (54.21+/-6.34)%, (58.78+/-4.70)%, (48.24+/-6.75)% respectively for IL-10, TGF-beta1 and LPS induction (P<0.01), but there was no significantly different among the three groups (P>0.01). The semi-quantitative RT-PCR and Western blot revealed that IL-10 and TGF-beta1 induced the higher PIR-B level and lower PIR-A level. On the contrary, the LPS down-regulated the PIR-B expression and up-regulated the PIR-A expression. Realtime PCR examination demonstrated that PIR-A and co-stimulating molecules such as CD80, CD86 and MHC-II were increased significantly after stimulation with LPS. Compared with the DC2.4 cells and the LPS-DC, the T-DCs inhibited alloactivated T cell proliferation and down-regulated the IFN-gamma secretion in MLR supernatant. Si-DC promoted the T cell proliferation (P<0.01) and enhanced the IFN-gamma secretion (P<0.01). It was concluded that up-regulating the PIR-B and down-regulating the PIR-A expression were the general feature of phenotype and constructed the new targets for dendritic cells to acquire immune tolerance in mice. Overexpression of PIR-B can inhibit the up-regulation of the PIR-A, CD80, CD86 and MHC-II expression, which might be the molecular mechanism for the T-DC.

Objective To investigate the expression of macrophage migration inhibitory factor(MIF) in renal tissue of children with Henoch-Schonlein purpura nephritis(HSPN),and its correlation with clinical indexes and pathological changes,and to explore its effect on the pathogenesis of HSPN.Methods According to the clinical manifestation,60 children with HPSN were divided into only purpura group,mixed group and HSPN group.MIF concentration of Henoch-Schonlein purpura(HSP) groups and healthy control group were detected with enzyme linked immunosorbent assay(ELISA).MIF protein expression and the marker of human macrophage(CD68) in renal tissues of HSPN and normal control group were detected with immunohistochemistry method.The total urine protein for 24 hours and urinary N-acetyl-beta-D-glucosaminidase (NAG) level were detected with laboratory routine method.Results MIF concentration in mixed group and HSPN group were significantly higher than that in only purpura group and healthy control group(Pa

AlM: To analyze personality traits in preoperative patients who undergolaser in situ keratomileusis ( LASlK) and to provide psychological basis for the selection of refractive surgery.
METHODS: Eligible patientswere seeking customized LASlK (group A n=53), conventional LASlK(group B n=75)and non-operation patients with ametropia (group C n=71 ) , who completed 16 personality factor questionnaires (16PF). Statistical analyses were performed with one-way ANOVA by SPSS11. 0 software package.
RESULTS: Compared to group C, patients in group A scored high on dominance and tension levels, and low on emotional stability level(P<0. 05), while patients in group B were more venturesome and more experimenting, but were less vigilant ( P<0. 05 ). Additionally, there were statistical differences between group A and group B on dominance, gallantry and vigilance respectively(P<0. 05). CONCLUSlON: The data indicates that personality profiles of LASlK patients with refractive error influence their decision for correction. Patients need suitable psychological assessment before surgery who actively chose customized LASlK seem to be more assertive and suspicious.

Objective To investigate the effect of small interfering RNA (siRNA) targeting cytoplasmic domain of tissue factor on apoptosis of vascular endothelial cells. Methods Specific siRNA targeting cytoplasmic domain of tissue factor were designed, and synthetic oligos were inserted into plasmid DNA. The siRNA constructs were transfected into human umbilical vascular endothelial cells (HUVEC) with liposome. The HUVEC were transfected with the constructs encoding siRNA Ⅰ, siRNA Ⅱ and pcDNA~(TM)6.2 GW/-miR plasmid separately. The transfected HUVEC were mixed with CD8~+ T lymphocytes. The apoptotic rate of tranfected HUVEC mixed with lymphocytes was analyzed by flow cytometry. Magnetic beads were used to measure PT of the supematant in the mixed lymphocytes culture. Results The siRNA constructs were confirmed by DNA sequence analysis. The apoptotic rate of HUVEC transfected with siRNA Ⅰ and Ⅱ plasmids was decreased significantly as compared with the empty control group (P<0.01). The apoptosis rate of HUVEC transfected with siRNA Ⅰ plasmid was lower than that of HUVEC transfected with siRNA Ⅱ plasmid (P<0.05). APTT of the culture supernatants in the three transfection groups was lower in the control groups (P <0.05), but there was significant difference among the three transfection groups. Conclusion The siRNA targeting cytoplasmic domain of tissue factor were successfully constructed, siRNA can protect HUVEC, and reduce the apoptotic rate of endothelial cells in mixed lymphocyte reaction without influencing the coagulation function.

The effect of CO2 and the manner of CO2 offer on the growth rate and maximual cell density of ultro-high density culture of Chaetoceros mulleri in the photobioreactor were studied in the work. The amount of CO2 offered to the culture was controlled by the parameter of pH value in the culture. Furthermore the growth kinetics of Chaetoceros muller in the photobioreactor was studied. The results showed requirement of CO2 by the cells and the increase of pH in the culture were the key limiting factors to the growth, when a high cell concentration in the culture was reached. The offer of CO2 could improve the statute of CO2, could control the pH in the culture and increase the growth rate and maximum cell density. The results from the experiments of CO2 offer manner showed different efficiency to growth was resulted from differences of CO2 offer manner. The best way is mixing the CO2 and air before the CO2 was offered to the culture.
Bioreactors ; Carbon Dioxide ; pharmacology ; Culture Media ; Culture Techniques ; methods ; Diatoms ; growth & development

ERα36 is a novel subtype of estrogen receptor alpha (ERα) known to play an important role in breast cancer development and widely expressed in normal tissues and cells including nerve cells. However, the expression and function of ERα36 in nerve cells have not been well elucidated. To examine whether ERα36 is involved in differentiation of nerve cells, the differentiated and undifferentiated PC12 (PC12D and PC12unD) cells were used. Transfection of ERα36-shRNA plasmid into PC12 cells was performed to establish the ERα36 gene knock-down cells model. Immunocytofluorescence and Western blot were used to analyze the expression of Nestin, β-tubulinIII and Neu-N in the PC12 cells. The results showed that ERα36 was expressed in both cell types. Compared with PC12D cells, PC12unD cells showed higher expression of Nestin and lower expression of β-tubulinIII. ERα36-shRNA-mediated knock-down of ERα36 expression enhanced the expression of β-tubulinIII and Neu-N, but attenuated Nestin expressions in PC12unD cells; ERα36 knock-down in PC12D cells mediated Nestin, β-tubulinIII and Neu-N in a contrary manner. These results indicate that ERα36 knock-down appear to be associated with inhibiting differentiation in differentiated cells and promoting differentiation in undifferentiated cells, suggesting that ERα36 is a dual regulator in nerve differentiation.
Animals ; Antigens, Nuclear ; metabolism ; Cell Differentiation ; Estrogen Receptor alpha ; genetics ; metabolism ; Gene Knockdown Techniques ; Nerve Tissue Proteins ; metabolism ; Nestin ; metabolism ; Neurons ; cytology ; metabolism ; PC12 Cells ; Rats ; Transfection ; Tubulin ; metabolism

BACKGROUNDThere is no agreement as to whether intensive glucose control in type 2 diabetes can reduce the incidence of macrovascular events in these patients. We performed a meta-analysis comparing intensive glucose control or conventional glucose control in randomized controlled trials.

METHODSDatabases including MEDLINE, EMBASE, and Cochrane controlled trials register, the Cochrane Library, and Science Citation Index were searched to find relevant trials. Outcome measures were the incidence of major macrovascular events.

RESULTSSix trials involving 28 065 patients were included. Analysis suggested that there was an obviously decreased incidence of major macrovascular events in patients having intensive glucose treatment vs. controls (RR 0.92; 95%CI 0.87, 0.98; P = 0.005). However, intensive glycemia control strategies in type 2 diabetes showed no significant impact on the incidence of death from any cause compared with conventional glycemia control strategies, intensive 14.7%, controls 12.0% (RR 0.95; 95%CI 0.80, 1.12; P = 0.55), as well as on the incidence of cardiovascular death, intensive 3.7%, controls 3.6% (RR 1.10, 95%CI 0.79, 1.53; P = 0.57).

CONCLUSIONSControl of glycemia to normal (or near normal levels) in type 2 diabetes appears to be effective in reducing the incidence of major macrovascular events, but there were no significant differences of either the mortality from any cause or from cardiovascular death between the two glycemia-control strategies.

Blood Glucose ; analysis ; Diabetes Mellitus, Type 2 ; blood ; complications ; drug therapy ; mortality ; Diabetic Angiopathies ; prevention & control ; Glycated Hemoglobin A ; analysis ; Humans ; Randomized Controlled Trials as Topic

OBJECTIVETo study the response of rat bone marrow mesenchymal stem cells (MSCs) to a single period of mechanical strain and expression patterns of transforming growth factor-beta (TGF-beta) and insulin-like growth factor-II (IGF-II) after mechanical stretch.

METHODSBone marrow MSCs were isolated from SD rats and cultured in vitro. A four-point bending apparatus were used to perform a single period of mechanical strain (2000 microepsilon, 40 min) on MSCs. Cellular proliferation and alkaline phosphatase (ALP) activity of MSCs were examined and gene expression patterns of TGF-beta and IGF-II were detected by SYBR green quantitative real-time RT-PCR.

RESULTSCell proliferation, ALP activity and expression of TGF-beta and IGF-II were all significantly up-regulated in stretched MSCs when compared with their controls. The mRNA levels of TGF-beta and IGF-II got top increase immediately after mechanical loading and increased about 51.44 and 8.92 folds, respectively, when compared with control cells. Expression of TGF-beta and IGF-II decreased with time and returned to control level at 12 h after mechanical stimulus, despite of a small increase at 6 h.

CONCLUSIONThe mechanical stretch can promote MSCs proliferation, up-regulate its ALP activity and induce a time-dependent expression increase of TGF-beta and IGF-II which in turn result in osteogenic differentiation of MSCs. Mechanical stimulus is a key stimulator for osteogenic differentiation of MSCs and vital for bone formation in distraction osteogenesis.

Animals ; Bone Marrow Cells ; Cell Differentiation ; Cell Proliferation ; Insulin-Like Growth Factor II ; Mesenchymal Stromal Cells ; Osteogenesis ; Osteogenesis, Distraction ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Somatomedins ; Transforming Growth Factor beta