Objective To evaluate the feasibility of composite material of bone morphogenetic protein (BMP) and PLGA/TCP in repair of peri-porous-titanium bone defects in adult rabbits.Meth- otis The composite of PLGA/TCP scaffold with bovine BMP was made and implanted in distal bone de- fects peri-porous-titanium in femur of adult rabbits.The effect of BMP with PLGA/TCP on peri-prosthesis was evaluated by means of scanning electron microscope (SEM),energy dispersive X-ray analysis (EDX) and biomechanics.Results BMP with PLGA/TCP showed obvious peak of Ca and P of EDX in the pores of titanium in experiment group six weeks after operation and higher than that in control group (P＜0.05).Push-out test demonstrated that the bonding strength between the composite of HA coating/ porous titanium and bone was increased significantly with time (P＜0.05).In experiment group,at 6 and 12 weeks,peri-porous-titanium had higher shearing force compared with control group (P＜0.05). Conclusion BMP loaded with PLGA/TCP is a promising bone graft for bone defects in revision arthro- plasty,as indicates that bone induction of BMP plays an important role in biological stabilizaiton.

OBJECTIVETo investigate the quantity and function of circulating dendritic cells (DC) in patients with chronic idiopathic thrombocytopenic purpura (ITP).

METHODSHigh dose dexamethasone (HD-DXM) at a dose of 40 mg orally per day for four consecutive days was the initial treatment for chronic ITP patients. Flow cytometry was used to analyze the number of myeloid DC (mDC), plasma cytoid DC (pDC) and CD4+FOXP3+ T cells in patients before and after the treatment, meanwhile the co-stimulatory molecules on circulating DCs were assayed as well. Monocyte-derived DCs and CD4+ T cells were co-cultured with autologous or allogeneic normal fresh platelets and after 6 days of incubation H-TdR was used to assay the proliferation of CD4+ T cells.

RESULTSThe absolute numbers of circulating mDC and pDC were not significantly different between pre-treatment patients and healthy controls (P > 0.05 and P >0.05). However, percentage of CD4+ FOXP3+ T cells was decreased (P < 0.01), and their percentage was inversely correlated with the number of pDC and mDC (r = -0.396, P =0.045 and r = -0.410, P =0.037). The initial response rate to HD-DXM was 92.3%. After 4-days treatment, CD4 FOXP3+ Treg cells increased (P <0.01) while pDCs decreased (P <0.01). Although mDCs increased after HD-DXM (P <0.05), their CD11c expression level was decreased (P < 0.01), the mean fluorescence intensity (MFI) decreased from 340 +/- 30 before treatment to 199 +/- 21 after treatment. The inverse correlation between pDCs and CD4+ FOXP3+ Treg cells remained (r= -0.524, P =0.006) while that between mDCs and Treg cells disappeared (r = - 0.360, P =0.071). The MFI of CD86 on DCs was higher in ITP patients than in healthy controls (P <0.05), while the proportions of CD86, CD40, CD80 and the MFI of CD40, CD80 in ITP patients were normal (P > 0.05). DCs from chronic ITP patients co-cultured with autologous or allogeneic platelets were highly efficient in stimulating autologous CD4+ T cells proliferaton as compared to those derived from healthy donors (P < 0.05 and P <0.05).

CONCLUSIONDCs may play a role in the pathogenesis of chronic ITP in relation with CD4+CD25+ Treg cells.

Adult ; CD4-Positive T-Lymphocytes ; immunology ; Dendritic Cells ; immunology ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood ; immunology

Humans ; Nontuberculous Mycobacteria ; Peritoneal Dialysis/adverse effects* ; Peritonitis/etiology*

OBJECTIVETo investigate the role of CD4+ CD25+ regulatory T (Treg) cells in patients with ITP.

METHODSFlow cytometry was used to examine the number of CD4+ CD25+, CD4+ CD25 high, CD4+ Foxp3+ and CD4+ CD25+ Foxp3+ T cells. The level of Foxp3 mRNA expression was analyzed by realtime quantitative reverse transcriptase polymerase chain reaction (RT-PCR) . CD4+ CD25 high T cells was cocultured with CD4+ CD25 - T cells from patients or controls for assessing the regulatory properties of CD4+ CD25 Treg cells.

RESULTSThe proportion of CD4+ CD25+ T cells in the peripheral blood of patients with ITP [(15.64 +/- 5.82) %] was significantly higher than that in normal control group [(9.30 +/- 3.95)%] (P <0.01). There was no significant difference in the percentages of CD4+ CD25 high T cells between ITP patients and controls [(1.53 +/- 0.66)% versus (1.36 +/- 0.55)% (P = 0.317)]. But the number of CD4 Foxp 3+ T cells and CD4 + CD25+ Foxp3+ T cells in patients were significantly lower than that in control group (P <0.01). The expression of Foxp3 mRNA reduced (P < 0.01) and the suppressive activity of CD4+ CD25 high T cells is lower than that of healthy controls (P <0.01).

CONCLUSIONIn patients with ITP, both the number and immuno-regulative function of CD4+ CD25+ Treg cells are reduced.

Adolescent ; Adult ; Aged ; Female ; Forkhead Transcription Factors ; metabolism ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; metabolism ; RNA, Messenger ; metabolism ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo study transplantation of mouse bone marrow mesenchymal stem cells (MSCs) into the xenogeneic testis.

METHODS(1) The tibias and femurs were dissected from 5-6-week-old mice. The marrow in the tibias and femurs was flushed out with medium. MSCs were isolated, cultured and purified in vitro by Percoll density gradient centrifugation combined with adherent method. (2) MSCs of the third generation were adopted and marked with Hoechest33342 for observation, and then made into cell-suspending fluid. (3) The marked MSCs were transplanted into the testis of the xenogeneic mouse by testis net injection. The biopsies of the testis tissues were carried out at different time and made into frost slices at three sites for observation.

RESULTS(1) A lot of purified MSCs were obtained at the third generation. (2) The nucleoli of the marked MSCs showed light-yellow under the fluoroscope. (3) Xenogeneic transplantation of mouse bone marrow MSCs by testis net injection was successful, without immunoreaction. On the 1 st day after transplantation, MSCs only concentratively distributed in the medial slices, the nucleoli being light-yellow; On the 1 st and 3 rd day, MSCs dispersively distributed in the medial slices; On the 6th, 9th and 12th day, MSCs presented in all the slices of the three sites, some ranging tubally; On the 15th and 18th day, the fluorescence of MSCs weakened; On the 21 st day, the fluorescence of MSCs disappeared.

CONCLUSIONTransplantation of mouse bone marrow MSCs into the xenogeneic testis by testis net injection is effective and feasible, without immunoreaction. MSCs can survive after transplantation.

Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Graft Survival ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred BALB C ; Testis ; surgery

OBJECTIVETo explore the eukaryotic expression of arresten in CHO cells and to investigate its basic biological activities.

METHODSCHO cells were divided into three groups: transfected pSecTag-arresten group, transfected pSecTag group and control group without transfection. PSecTag-arresten was transfected into CHO cells by Lipofectamine 2000 method. The arresten mRNA in CHO cells was assayed by RT-PCR. The protein expression of arresten gene was examined by Western-Blot. The cells expressing arresten were screened out by Zeocin. The effect of arresten on huvec cell migration and anchoring to three-dimensional vascular structures was measured.

RESULTSThe result of RT-PCR and Western-blot showed that arresten gene has been successfully transfected into CHO cells and expressed in those cells. Arrssten inhibited huvec cell migration and anchoring to three-dimensional vascular structures.

CONCLUSIONCHO cells expressing arresten have been obtained successfully. Arresten can inhibit huvec cell migration and anchoring to three-dimensional vascular structures, indicating that it might be one of its anti-angiogenetic approaches.

Angiogenesis Inhibitors ; biosynthesis ; genetics ; pharmacology ; Animals ; Blotting, Western ; CHO Cells ; Cell Line ; Cell Movement ; drug effects ; Cells, Cultured ; Collagen Type IV ; biosynthesis ; genetics ; pharmacology ; Cricetinae ; Cricetulus ; Endothelial Cells ; cytology ; drug effects ; physiology ; Humans ; Neovascularization, Physiologic ; drug effects ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo investigate the effect of ecdysterone (EDS) on the proliferation of human bone marrow mesenchymal stem cells (hMSCs) in vitro.

METHODShMSCs were isolated from human bone marrow cell suspension by density gradient centrifugation. The expression of integrins CD44, CD105, CD34 and CD29 were examined by immunocytochemical method. EDS at 10, 25, 50 or 100 microg/ml were added in hMSC culture system, using the routine culture medium for hMSCs as control. The cell viability were analyzed by MTT assay and the cell cycle changes were examined by flow cytometry.

RESULTSThe optical density (OD) differed significant between the EDS treatment groups and the control group (P<0.01), and 25 microg/ml EDS group showed the highest OD value (P<0.01) without significant differences among 10, 50 and 100 microg/ml EDS groups (P>0.05). Flow cytometry showed that treatment of the cells with 25 microg/ml EDS significantly increased the cell percentages in S and G(2)M phases and the proliferation index (PI) of the cells as compared with the control group.

CONCLUSIONWithin a given concentration range, EDS can promote the proliferation of hMSCs in vitro, and this effect can be the most obvious at the concentration of 25 microg/ml. The effect of EDS in promoting the proliferation of hMSCs does not positively correlate to EDS concentration administered.

Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Ecdysterone ; pharmacology ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology

Long duty hours have been associated with significant medical errors, adverse events, and physician “burn-out”. An innovative night float (NF) system has been implemented in our internal medicine program to reduce the negative effects of long duty hours associated with conventional full-call systems. However, concerns remain if this would result in inadequate training for interns. We developed a structured questionnaire to assess junior doctors’ perceptions of the NF system compared to full calls, in areas of patient safety, medical training, and well-being. Ninety-seven (71%) of the 137 doctors polled responded. Ninety-one (94%) felt the NF system was superior to the full call system. A strong majority felt NF was beneficial for patient safety compared to full call (94% vs. 2%, p<0.001). The NF system was also perceived to reduce medical errors (94% vs. 2%, p<0.001) and reduce physician “burn-out” (95% vs. 5%, p<0.001). Beyond being a practical solution to duty-hour limitations, there was a significant perceived benefit of the NF system compared to the full call in terms of overall satisfaction, patient safety, reducing medical errors and physician “burn-out”.
After-Hours Care ; Asian Continental Ancestry Group ; Education, Medical ; Humans ; Internal Medicine ; Medical Errors ; Patient Safety ; Patient Satisfaction

Objective To discuss the clinical value of transabdominal sonography after bowl preparation in diagnosis of ileocecal valve syndrome ( IVS) .Methods The ultrasonic features of IVS in 37 cases were summerized and correlated with the follow-up findings after conservative treatment or the pathologic results after operation .Twenty-eight cases were confirmed by follow-up and 9 cases by operative pathology.Results Among the 37 cases of IVS,28 were idiopathic IVS (75.7%,28/37) and 9 were secondary IVS (24.3%, 9/37%).For the secondary cases, the primary diseases included 5 acute appendicitis,2 Meckel diverticulum,1 terminal ileitis and 1 carcinoma of ascending colon .The diagnostic accuracy rate of ultrasound was 89.2%(33/37).Misdiagnosis rate was 10.8%(4/37),including 1 case of idiopathic and 3 cases of secondary IVS .The IVS ultrasonic images coulde be displayed clearly using 7.0-10.0 MHz probes.In fasting examination,three ultrasonic characteristic signss were found in interminal ileum region at the right lower abdomen .And these features were bagel-shaped sign [91.9%(34/37),average size (1.9 ±1.6) cm ×(0.8 ±0.3) cm],short sleevelet-shaped sign [91.9% (34/37,average size (2.1 ± 0.4)cm ×(1.3 ±0.2) cm],and rose-shaped sign [83.8% (31/37),average size (1.4 ±0.2) cm × (1.0 ±0.2) cm].The shapes of some signs were changeable when the probe compressed .In the case of idiopathic IVS ,several pathologic changes could be seen on sonography after intestinal tract filling of oral 20%mannitol,including slight thickened mucosa and submucosa of erminalileum ,enlarged ieocecal valve and the crocodile-mouth sign.Conclusions Transabdominal ultrasonic examination with high frequency probe after bowl preparation plays an important role in diagnosis of IVS .The method is simple and accurate and should be recommended and applied clinically .

Long duty hours have been associated with significant medical errors, adverse events, and physician “burn-out”. An innovative night float (NF) system has been implemented in our internal medicine program to reduce the negative effects of long duty hours associated with conventional full-call systems. However, concerns remain if this would result in inadequate training for interns. We developed a structured questionnaire to assess junior doctors’ perceptions of the NF system compared to full calls, in areas of patient safety, medical training, and well-being. Ninety-seven (71%) of the 137 doctors polled responded. Ninety-one (94%) felt the NF system was superior to the full call system. A strong majority felt NF was beneficial for patient safety compared to full call (94% vs. 2%, p<0.001). The NF system was also perceived to reduce medical errors (94% vs. 2%, p<0.001) and reduce physician “burn-out” (95% vs. 5%, p<0.001). Beyond being a practical solution to duty-hour limitations, there was a significant perceived benefit of the NF system compared to the full call in terms of overall satisfaction, patient safety, reducing medical errors and physician “burn-out”.