[Objective]To evaluate the effect of bone mesenchymal stem cells(BMSCs) on healing of tendon-bone junction in a bone tunnel and to provide experimental evidence of labeling and tracing the stem cell.[Method]BMSCs from rats were harvested by adherent separation screening method and labeled by SPIO and DiI.Thirty-nine 8-week-old rats were randomly divided into two groups,21 rats injected with double-labeled BMSCs and Pluronic F-127 as experimental group and 18 rats injected with Pluronic F-127 alone as control group.In each group,biomechanical analysis was carried out to assess the effect of healing at 2,4 and 8 weeks.The transplanted BMSCs were traced by fluorescence microscope at 2,4,8 weeks and 7.0T MR instantly,3 and 7days after operation.[Result]BMSCs were labeled effectively by SPIO and DiI.DiI-labeled positive cells were observed between tendon and bone with fluorescence microscope at 2,4 and 8 weeks.No significant signal changes of tendon-bone interface could be observed by 7.0 T MR.Biomechanically,maximum pullout load had no statistical difference between experimental group and control group at 2 weeks.At 4 and 8 weeks,the experimental group had significantly higher maximum pullout load compared with control group(P

Objective To explore the effect of prevention and treatment of external application of Algoplaque for controlling phlebitis caused by peripheral indewelling needle in vein for patients. Methods This research was divided into two parts,prevention and treatment. As for prevention research,patients were randomly divided into the experimental and the control groups,each group included 30 patients. In the experimental group,we applied directly external application of Algoplaque at the upper of needle puncture site of the vein and nearby the eye. In the control group,we applied the film directly to fix the indwelling needle. As for the treatment research, it was carried out in patients with occurred phlebitis, who were randomly divided into two groups,the experimental group included 30 cases of patients and the control group included 28 cases of patients. Observation time was one to five days. Results The incidence of phlebitis in the experimental group of prevention research was 23%, in the control group it was 90%. The incidence of phlebitis in the experimental group was significantly lower than that of the control group. The effective rate in the experimental group of treatment research was 96.7% and it was 67.9% in the control group. The difference was very significant. Conclusions External application of Algoplaque can effectively control phlebitis caused by peripheral indewelling needle in vein.

To express Pleurocidin in Escherichia coli and to enhance the secretory efficiency of the fusion protein, the gene encoding Pleurocidin was ligated with Cherry DNA sequence via blunt-end ligation. Then this fusion gene was cloned into pET22b (+) vector and the recombinant plasmid was transformed into E. coli BL21 (DE3). Lactose was used to induce expression of fusion protein. The recombinant plasmid pET22b (+) -CP was successfully constructed and high-level expression of fusion protein was induced with lactose. Statistics showed that addition of glycine after 16 h of induction significantly enhanced the secretory efficiency of the fusion protein. After hydrolysis of the fusion protein by diluted hydrochloric acid and some further purification steps, r-Pleurocidin was obtained with antibacterial activity against E. coli DH5α and Bacillus subtilis BS168. In conclusion, the fusion protein was expressed in E. coli and biologically active r-Pleurocidin was obtained after hydrochloric acid cleavage and purification.
Animals ; Cloning, Molecular ; Escherichia coli ; metabolism ; Fish Proteins ; biosynthesis ; Flounder ; Recombinant Fusion Proteins ; biosynthesis

AIM:To investigate the role of mesenchymal stem cell-induced regulatory dendritic cells ( MSC-DCregs) in mouse acute graft-versus-host disease( aGVHD) model.METHODS: Bone marrow cells from BALB/c ( H-2 d ) mice were isolated and were induced to differentiate into DCs.The DCs were selected by flow cytometry, and after 10 d co-culture with MSCs, they were induced to be MSC-DCregs.Male 8-week-old C57BL/6 (H-2b) mice were used as do-nor mice.The female 8-week-old BALB/c (H-2d) mice, who had received 100 cm source-skin distance, 30 cm ×30 cm radiation field, 700 cGy total body irradiation (TBI) pretreatment were used as recipient mice.The recipients were divided into 5 groups:control group, TBI group ( injected with medium only) , bone marrow transplantation group ( injected with 1 ×107 bone marrow cells), aGVHD group (injected with 1 ×107 bone marrow cells and 1 ×107 spleen cells), and MSC-DCregs group (injected with 1 ×107 bone marrow cells, 1 ×107 spleen cells and 1 ×106 MSC-DCregs).The white blood cell count, recipients’ chimerism, clinical evaluation of aGVHD, survival analysis and pathological changes were deter-mined.RESULTS:Hematopoieic recovery was seen at 10 d after transplantation.The recipients’ chimerism was parallel to the donors’ at 30 d.The median survival time of the mice in aGVHD group and MSC-DCregs group was 27 d and 33 d, and the survival rates at 30 d were 20% and 100% (P<0.01), respectively.The clinical scores of the mice in MSC-DCregs group were lower than those in aGVHD group ( P<0.01) .Moreover, the pathological changes in the skin and liver of the mice in MSC-DCregs group were less serious than those in aGVHD group.CONCLUSION: The MSC-DCregs in-duce an aGVHD tolerance in vivo, and further research of its mechanism is still in great necessary.

AIM:To investigate the expression and potential role of complement 5a receptor (C5aR) in chro-nic graft-versus-host disease (cGVHD).METHODS:The expression of C5aR on lymphocytes and the frequency of CD4+CD25+ Foxp3+ regulatory T cells (Tregs) in 20 cGVHD patients and 9 healthy donors was detected by flow cytometry.The correlation between the expression of C5aR and the percentage of Tregs in the cGVHD patients was analyzed.In addition, the splenocytes from the mice were cultured in vitro, and stimulated these splenocytes with recombinant mouse C5a protein (rmC5a).The peripheral blood mononuclear cells (PBMCs) from cGVHD patients were cultured in vitro, which was inhibited by C5aR antagonist (C5aRA).The frequency of Tregs in these splenocytes and the PBMCs were evaluated by flow cytometry.RESULTS:The expression of C5aR on the lymphocytes was significantly increased in the cGVHD patients compared with the healthy donors, while the percentage of Tregs was markedly lower in the cGVHD patients.The expression of C5aR was negatively correlated with the percentage of Tregs.Furthermore, the development of Tregs was suppressed by rmC5a stimulation, but was promoted by C5aRA in vitro.CONCLUSION:C5aR elevation is associated with Treg reduction in cGVHD, indicating that C5aR may play a potential role in suppressing Tregs, resulting in the incidence of cGVHD.

Professor 's experience of acupuncture combined with medication for epilepsy is summarized, which is explained from epilepsy's etiology and pathogenesis, diagnosis and treatment of acupuncture and medication, respectively. Besides, the theoretical foundation and use instruction of acupuncture technique "-" for epilepsy are introduced. Professor highly values the adherence to etiology and pathogenesis, pays attention to syndrome differentiation and searches for the primary disease cause. He proposes the wind, phlegm, stasis and deficiency are the pathogenesis of epilepsy, and points out acupuncture could be applied during attack stage and remittent stage, but electroacupuncture should be used with caution. Regulating spirit is the key for treating epilepsy. The combination of acupuncture and medication could regulate the governor vessel and guide to the origin, which have significant curative effect.
Acupuncture Therapy ; Combined Modality Therapy ; Electroacupuncture ; Epilepsy ; therapy ; Humans