Objective The purpose of this study was to develop a new cultural method, so that the rat bone marrow stromal cells (MSCs) can well differentiate into osteoblasts in vitro and the cultured osteoblasts can be identified. Methods MSCs of rats were isolated and cultured, then the cultured cells were identified with inverted microscopy and undergone an HE stain for observing the morphology of living cells, and the cells were histochemically stained for examining the activity of cellular alkaline phosphatase (ALP), as well as were radio-immunologically analyzed for detecting the secretion of osteocalcin. Results The MSCs were typical fibroblast-like and possess a strang proliferative capability after cultured in vitro.The cellular ALP activity and osteocalcin secretion achieved the peak value from the fortieth day and maintained the status until the sixtieth day after cultured in vitro.Conclusion The data demonstrated that the new developed culture method has the advantages of short time, less risk of contamination and higher efficiency, and conducive to MSC's differentiating and proliferating into osteoblasts that do function well.

Objective:Because of the lower proliferation ability of periodontal ligament cells of elderly people,how to choose suitable scaffold materials for them becomes very important.The aim of this study was to observe the morphological manifestation of the periodontal ligament cells of elderly people seeded onto the three-dimensional porous nano-hydroxylapatite scaffolds.Methods: Periodontal ligament cells from elderly people were seeded onto the three-dimensional porous nano-hydroxylapatite scaffolds,cultured for 15 days in vitro,and observed for the morphological changes by scanning electron microscopy in comparison with those of young people.Results:Fifteen days after seeded on the three-dimensional porous nano-hydroxylapatite scaffolds,the periodontal ligament cells of the elderly people exhibited sufficient adhesive behavior,proliferated well and produced tiny calcium nodules,but the number of the nodules was smaller than that in the young people.Conclusion: The periodontal ligament cells of elderly people grow well on three-dimensional porous nano-hydroxylapatite scaffolds,and can be considered as a suitable scaffold for periodontal tissue engineering in elderly people.

Objective:To investigate the feasibility of using marrow stromal cells (MSC) as seed cells and three-dimensional porous nano-hydroxylapatite as scaffolds for constructing tissue-engineered bone.Methods:MSC from rat were cultured and induced to differentiate into osteoblasts in vitro. And then these induced cells were identified and seeded onto three-dimensional porous nano-hydroxylapatite scaffolds, cultured for 15 days in vitro. Scanning electron microscopy was used to evaluate the growth of these induced cells on three-dimensional porous nano-hydroxylapatite scaffolds. Results: The activity of alkaline phosphatase (ALP) and the secretion of osteocalcin of MSC from rat appeared and were increased gradually along the culture. The cells seeded on three-dimensional porous nano-hydroxylapatite scaffolds could adhere and proliferate well, and come into being many tiny calcium nodules and collagenous fibers. Conclusion:The data demonstrated that the new developed culture method is conducive to MSC’s differentiating and proliferating into osteoblasts that have a fine activity and the three-dimensional porous nano-hydroxylapatite may be considered as a suitable scaffold for the seeded cells. Using marrow stromal cells as seed cells and three-dimensional porous nano-hydroxylapatite as scaffolds is advantageous for constructing tissue-engineered bone.

Objective To observe effects of chitosan on proliferation and differentiation of human periodontal ligament cells( PDLC). Methods The human periodontal ligament cells were isolated and cultured.The effects of chitosan on proliferative ability of human PDLCs were evaluated with MTT method, and alkaline phosphatase activity and the secretion of osteocalcin of human PDLCs were measured with spectrophotometric assay and radioimmunological method. Results Compared with control group, the proliferative ability of human PDLCs in Chi(0.05g?L~ -1) group and Chi(0.1g?L~ -1) group at 3,5,7 day was considerably increased (P

?Objective:To investigate the behavior of absorbable coral hydroxyapatite (Interpore 500R) as bone tissue engineering scaffold. Methods: Marrow stromal stem cells harvested from legally aborted human fetus were cultured in DMEM containing 100 ml/L fetal boving serum and induced to differentiate towards osteoblasts.Then the cells were seeded onto absorbable coral hydroxyapatite , then the composite was implanted subcultaneously into the back of nude mice .Cell materia attachement was observed with scanning electron microscope,type I collegen secretion was studied with laser confocal microscope and new bone formation was studied histologicaley. Results: The cells could adhere to absorbable coral hydroxyapatite and secrete collagen I.Mature bone tissue was found in the implanted composite 40 day after transplantation. Conclusion: Absorbable coral hydroxyapatite Interpore 500R is a favorable bone tissue engineering scaffold.

Objective To evaluate the efficacy of Huangqi inoculation fluid union chemotherapy in the treat- ment of the induction chemotherapy time of acute leukemia.Methods The patients with acute leukemia were ran- domized into two groups:the 1st group treated with conventional induction chemotherapy(control-group) and the 2nd group treated with chemotherapy treatment union Huangqi inoculation fluid(observation-group).Chinieal effects and poisonous side effects of two groups were compared.Results The completely alleviating rate of observation- group was 87.75%,the partially alleviating rate of observation-group was 4.04%,and KPS grades total improve- ment rate was 73.47%;the completely alleviating rate of control-group was 50.00%,the partially alleviating rate of control group was 16.67%,and KPS grades total improvement rate was 36.67%;the poisonous side effects of ob- servation-group was lower titan that of the control-group(P

AIM To investigate the possible role of the mitochondrial transmembrane potential (??m) and caspase 3 in arsenic trioxide(As 2O 3)-induced apoptosis of cancer cells. METHOD Namalwa, SGC7901 and Bcap37 cell lines were used as in vitro models. Apoptosis was confirmed by sub-G 1 cells content as well as phosphatidylserine(PS) externalization. The ??m was detected on flow cytometry through double staining of Rhodamine 123(Rh123) and popidium iodide(PI). In addition, the effect of DEVD-CHO, a selective inhibitor of Caspase 3, on As 2O 3-induced apoptosis was studied. RESULT The As 2O 3-induced apoptosis closely associated with the externalization of the ??m and the activation of Caspase 3. As 2O 3 induced cells necrosis when Caspase 3 was inhibited. CONCLUSION As 2O 3 may selectively activates Caspase 3 after it induces externalization of ??m, which causes cancer cells apoptosis.

AIM:To investigate matrine-induced apoptosis of human medulloblastoma D 341 cells and the ex-pression of Bax , Bcl-2, serine/threonine kinase Akt and phosphorylated Akt ( p-Akt) in vitro.METHODS: D341 cells were divided into experimental groups ( added with matrine at different concentrations ) and control group ( under the same conditions without matrine).The proliferation of D341 cells was analyzed by CCK-8 assay.Apoptosis was detected by An-nexin V-FITC/PI double staining and the expression of Bax , Bcl-2, Akt and p-Akt was detected by Western blotting .RE-SULTS:Matrine significantly inhibited the proliferation of D 341 cells and increased the apoptosis in a dose-and time-de-pendent manner .The cell apoptosis was characterized by chromatin condensation with margination of chromatin to the nu -clear membrane , increased when and larger cytoplasmic vacuoles , and formation of apoptotic body after treatment with ma-trine.The expression of Bax increased , while the expression of Bcl-2 and p-Akt decreased when the drug concentration gradually increased .CONCLUSION:Matrine induces the apoptosis of human medulloblastoma D 341 cells in vitro by acti-vation of Bax, down-regulation of Bcl-2 and reduction of p-Akt expression level in the PI3K/Akt signaling pathway.

Immunoenhancement activity of bee pollen and its acetone extract was studied in normal, sarcoma-180 bearing, cyclophosphamide- and antilymphocyte serum-treated mice.Bee pollen and its acetone extract given orally for 30 days could significantly increase the production of serum anti-SRBC hemolysin (HC50) and the number of spleen plaque forming cells (PFC) in primary response to sheep red blood cell (SRBC) in young and adult mice. The acetone extract of bee pollen could significantly prevent the decrease of HC50) the number of PFC and specific rosette forming cells (SRFC), and the quantitative hemolysin of spleen cells (QHS) against SRBC in S-180 bearing, cyclophosphamide- and antilymphocyte serum-treated mice respectively.These results suggested that bee pollen of Brassica campestris L. and its acetone extract have immune-enhancement activity.

Objective To investigate the immunological pathogenesis of Kawasaki disease( KD)through examination of changes in the expression of suppressors of cytokine signaling 1(SOCS1)and SOCS3,helper T cells and CD4 + CD25 + regulatory T cells(CD4 + CD25 + Treg)in peripheral blood from children with acute KD. Methods Six-teen children[10 boys,6 girls,aged 1 - 2 years old,averaged(1. 6 ± 0. 3)years old]in the acute phase of KD(KD group),16 children[9 boys,7 girls,aged 1 - 3 years old,averaged(1. 5 ± 1. 1)years old]with pneumonia(pneumo-nia group)and 8 normal children[5 boys,3 girls,aged 1 - 5 years old,averaged(2. 0 ± 1. 1)years old]of the same age(normal control group)from the Affiliated Hospital of Qingdao University who were admitted from October 2012 to March 2013 were recruited. The mRNA levels of SOCS1 and SOCS3 in the T cells from peripheral blood were examined by way of reverse transcription - polymerase chain reaction(RT - PCR). Interferon - γ( IFN - γ),interleukin - 4 (IL - 4)and CD4 + CD25 + Treg were quantified by means of fluorescence activated cell sorting(FACS). Results The expressions of SOCS1 and SOCS3,the percentage of IL - 4 T cells observed in the peripheral blood of the pneumonia group were similar to the normal control group(P ﹥ 0. 05),but significantly decreased in the percentage of INF - γ and the level of CD4 + CD25 + Treg(t = 3. 71,12. 81,all P ﹤ 0. 05). Compared to the normal control group and the pneumo-nia group,the expressions of SOCS1 and SOCS3,the percentage of INF - γ and IL - 4 T cells decreased significantly in the peripheral blood of the KD group(t = 2. 27,4. 48,17. 64,2. 73,2. 74,1. 25,2. 36,2. 59,all P ﹤0. 05 ). On the other hand,the level of CD4 + CD25 + Treg in the peripheral blood of the KD group was markedly lower than that in the normal control group(t =7. 70,P ﹤0. 05),but similar to the pneumonia group(P ﹥0. 05). Conclusions The function of helper T cells is inhibited in acute KD. The CD4 + CD25 + Treg may be involved in the immunological pathogenesis of KD.