Based on M9 culture medium,the concentration of ingredients of culture medium was optimized for the fermentation of pNSR32/BL21(DE3),the engineering bacterial with spider silk protein,and lactose as an inducer.The condition of optimum culture medium was obtained for the expression of the high molecular weight recombinant spider silk protein by using orthogonal and individual factor experimental design.The result was showed that the optimum culture medium was consisted of 0.3% glycerol,3% yeast,0.75% tryptone,0.05%(NH_4)_2SO_4 and a little inorganic salt_.It was confirmed that the optimum culture medium will benefit the growth of bacterial and expression of recombinant spider silk protein.The production level of propose protein will attain at 20% of the total proteins in the fermentation.

Objective To compare the clinical efficacy of moxibustion Zhiyin with knee chest decubitus,ear acupoint magnetic bead press combined with knee chest decubitus and knee chest decubitus in correction of breech position,thus to provide more safe and effective correction method for the clinical treatment.Methods 150 pregnant women with breech were selected.They were randomly divided into observation group Ⅰ,observation group Ⅱ and control group according to the digital table,50 cases in each group.The control group was treated with knee chest decubitus to correct breech position,the observation group Ⅰ was given moxibustion Zhiyin combined with knee chest position,the observation group Ⅱ was given ear acupoint magnetic bead press combined with knee chest decubitus.Results The effective rates of the observation group Ⅰ and observation group Ⅱ were 96％,92％,respectively,which were higher than 76％ of the control group,the difference were statistically significant (χ2=5.09,4.18,all P＜0.05).The effective rate between the observation group Ⅰ and observation group Ⅱ showed no significant difference (P＞0.05).Conclusion Moxibustion Zhiyin combined with knee chest decubitus and ear acupoint magnetic bead press combined with knee chest decubitus in the correction of breech position have short course of treatment,high success rate,and are more economical and convenient,with high safety and good curative effect,which are worthy of promotion.

Objective To assess the osteogenic ability after co-culture BMSC and ADSC in vivo and in vitro.Methods ADSC and BMSC were obtained by adherent screening method and enzymatic digestion method.Flow cytometry was used to confirm the phenotypes of ADSC and BMSC.Oil red O was used to induce MSC to fat.Alkaline phosphatase (ALP) and alizarin red staining were used in osteogenic group.This sample was divided into four groups,no-induced stem cells group;BMSC osteogenic induction group;ADSC osteogenic induction group;co-culture of BMSC and ADSC osteogenic induction group.ALP activities and Calcium absorbance were determined during different periods of osteogenic introduction.OCN and Runx2 expression level were tested via RT-PCR and western blot methods after osteogenic induction for 2 weeks.Furthermore,cells in each group were seeded on HA/CS/PLLA composite scaffolds,and the scaffolds with cells were planted into bone defects in rat models.The rats were sacrificed by overdose anesthesia at 8 weeks after surgery and the scaffolds were removed for further analysis.Results Oil red O staining demonstrated red after adipogenic induction.Alkaline phosphatase and Alizarin red staining showed flaky red under condition of osteogenic induction.There had no statistical change among each group after osteogenic induction for 3 days,and ALP activity significantly increased after osteogenic induction for 5 days.Meanwhile,the ALP activity in co-culture of BMSC and ADSC group was markedly higher than the other three groups.However,there had no significant change in A value of calcium absorbance among each group after osteogenic induction for 7 days,while it increased at 14th day and ALP activity in co-culture of BMSC and ADSC group was significantly higher than the other three groups.After osteogenic induction for 2 weeks,the mRNA expression of OCN and Runx2 in co-culture of BMSC and ADSC group was 78.24±8.11 and 1 180.13±121.16 respectively,and the protein expression of OCN and Runx2 was 6.54±0.59 and 4.43±0.51.These mRNA and protein expression level in co-culture of BMSC and ADSC group enhanced significant compared with the other 3 groups.Histological assay demonstrated that the new bone tissues formed in co-culture of BMSC and ADSC group were 497.75±7.44 μm2,which was larger than that in the other 3 groups at 8 weeks after implantation.Conclusion Co-culture BMSC and ADSC may up-regulated the osteogenic ability in vivo and in vitro.

Objective To search for the methods of inducing the high stability rat middle cerebral artery occlusion (MCAO) model and to noninvasively assess the method of model effect. Methods Six kinds of filaments with different diameters were used to induce rat MCAO models and their success rate, incidence of subarachnoid hemorrhage, and infarct volume were compared. The model scores were performed to assess the model effect after the operation and at 24 hours after reperfusion. Results Higher model success rate and lower incidence of subarachnoid hemorrhage were achieved when the 0. 28 mm in diameter and 0.32-0.34 mm in tip diameter filaments were used. 1he sensibility and specificity were higher when the model scores were performed at 24 hours after reperfusion. Conclusions Using the filaments of 0.28 mm in diameter and 0. 32-0. 34 mm in tip diameter for intraluminal thread could improved the stability of the models. The model scores at 24 hours after reperfusion could noninvasively assess the model effect.

The probable mechanism of the reduction of rat cerebral ischemic-reperfusion injury by propyl gallate was studied. Intraluminal suture middle cerebral artery occlusion model of rat was employed. Propyl gallate was injected immediately after the ischemia was happened. The activity of NF-kappaB, and the expression of COX-2 and HSP70 on the peripheral ischemia were determined by Western blotting. The expression of TNF-alpha was determined by ELISA assay. RT-PCR and immunofluorescence staining were employed to detect the transcription and expression of TLR-4. Results showed that propyl gallate could inhibit the activity of NF-kappaB in the peripheral ischemia, and reduce the expression of COX-2 and TNF-alpha. As the upstream of NF-kappaB, the transcription and expression of TLR-4 decreased, as well as HSP70, the endogenic ligand of TLR-4. As an antioxidant, propyl gallate could reduce the cerebral ischemic-reperfusion injury through inhibiting the activity of NF-kappaB and decreasing the COX-2 and TNF-alpha in the peripheral ischemia. It also could influence HSP70 and TLR-4.

Aim The seminal plasma of 51 cases of infertile were studied with the PCR technice and ELISA method, of which 20 spectimens (39. 2％ )were Uu positive, 18 spectimens (35. 3 ％ ) were AsAb positive, positive of Uu and AsAb were 11 specimens(21.6％ ). Correlation of seminal plasma that there is significiant between thetwo groups in sperm activity score, sperm viability, liquefy time, deformative rate and sperm morphogenesis. The conclusion is that genital tract infection with Uu can affect semen parameters and diminish semen quality, the fertility ability of human sperm will be destroyed.

The myelodysplastic syndromes (MDS), which are characterized by the presence of ineffective hematopoiesis and an increased risk of transformation to acute myeloid leukemia (AML), are a group of clonal disorders deriving from damage of the hematopoietic stem/progenitor cells. The 58th American Society of Hematology (ASH) Annual Meeting consists of 5 main subjects, includingchronic myelomonocytic leukemia (CMML) and MDS biology and treatment, higher risk MDS clinical studies, lower risk MDS clinical studies, predisposition and diagnosis of MDS, and prognostic and predictive utility of recurrent somatic mutations in MDS. This article will introduce some highlights of the oral reports in this meeting.

ObjectiveTo observe the effect of megace (MA) on the survival condition of cancer patients in chemotherapy (CT) periods.Methods92 patients with cancer were divided into the MA group (treated with MA+CT) and control group (treated only with CT). The changing of the appetite, normal food amount, weight, gastrointestinal reaction and whole body conditions of two groups were evaluated.ResultsIn the MA group, 52.2% patients had appetite improvement, 47.8% had more food amount, 45.7% gained more weight, 50% had no obvious gastrointestinal reaction such as vomiting and nausea, and 50% had improve the survival condition according to the Karnofsky performance status (KPS) scores (increment >10). In the control group, only 6.5% had appetite improvement, 4.3% had more food amount, 13% gained more weight, 28.3% had no vomiting and nausea, and 15.2% had improved the survival condition. There was a significant difference between two groups (P<0.01).ConclusionMegace is able to reduce nausea and vomiting caused by CT, improve appetite, increase patients weight, protect bone marrow from the inhibition of CT, improve the life quality of cancer patients, and has no evident side-effects.

A series of noscapine analogues have been synthesized via 13-step reaction starting from 2-hydroxy-3-methoxybenzaldehyde. Anti-tumor activities of these compounds were evaluated against HL-60 cell lines in vitro by the standard MTT assay. It was found that most of these derivatives showed appreciable inhibitory activity against HL-60 and tubulin polymerization. The results also indicated that the potency of compound 31 is about three times more than that ofnoscapine against HL-60 cell line and tubulin polymerization. Moreover, it induced a massive accumulation of cells in G2/M phase. These results showed noscapine and its derivatives were worth to be intensively studied further.

Objective To elucidate the interaction between osteoblast of bone marrow microenvironment and leukemia cells,and to investigate the role of osteoblast in the leukemia cells survival and apoptosis and the influence of leukemia cells on the osteoblast.Methods Leukemia cells from AML1-ETO9a-Rac1 mouse leukemia model and osteoblast cells were used.The ratio of GFP+ leukemia cells that co-cultured with or without osteoblast was detected by FACS.In addition,the apoptosis level of leukemia cells was detected by flow cytometry by PI and Annexin Ⅴ labeling.Activation level of PARP was determined by Western-blot.Real-time PCR (RT-PCR) was utilized to detect the mRNA level of TPO,N-cadherin,OPN and Ang1 in osteoblast which was separated from leukemic mice.Results The ratio of GFP+ cells in AE9a-Rac1 leukemia cells co-cultured with osteoblast cell was significantly higher than that of AE9a-Rac1 leukemia cells cultured alone.The apoptotic level of AE9a-Rac 1 leukemia cells cultured alone was significant higher than that of AE9a-Rac 1 leukemia cells in co-culture system.Western blot showed that activated level of PARP in AE9a-Rac1 leukemia cells co-cultured with osteoblast was lower than that cultured alone.RT-PCR result showed that TPO and N-cadherin mRNA levels in primary osteoblast separated from leukemic mice were higher than that from normal mice.Ang1 and OPN mRNA levels of osteoblast from leukemia mice were lower.Conclusion Osteoblast cell can support the survival and inhibit the apoptosis of leukemia cells.Leukemia cells can influence the functions of osteoblast by microenvironment associated cytokines production.